Author: "Yann Guiguen" / Topic: male - Searchworks@Jio Institute Digital Library Search Results

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1. An ancient truncated duplication of the anti‐Müllerian hormone receptor type 2 gene is a potential conserved master sex determinant in the Pangasiidae catfish family

Author: Ming Wen, Qiaowei Pan, Elodie Jouanno, Jerome Montfort, Margot Zahm, Cédric Cabau, Christophe Klopp, Carole Iampietro, Céline Roques, Olivier Bouchez, Adrien Castinel, Cécile Donnadieu, Hugues Parrinello, Charles Poncet, Elodie Belmonte, Véronique Gautier, Jean-Christophe Avarre, Remi Dugue, Rudhy Gustiano, Trần Thị Thúy Hà, Marc Campet, Kednapat Sriphairoj, Josiane Ribolli, Fernanda L. de Almeida, Thomas Desvignes, John H. Postlethwait, Christabel Floi Bucao, Marc Robinson-Rechavi, Julien Bobe, Amaury Herpin, Yann Guiguen, Hunan Normal University (HNU), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Lausanne = University of Lausanne (UNIL), Plateforme Bio-Informatique - Génotoul, Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRAE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-BioInfOmics, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génétique Physiologie et Systèmes d'Elevage (GenPhySE ), Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-École nationale supérieure agronomique de Toulouse (ENSAT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génome et Transcriptome - Plateforme Génomique ( GeT-PlaGe), Plateforme Génome & Transcriptome (GET), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Génétique Diversité et Ecophysiologie des Céréales (GDEC), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Clermont Auvergne (UCA), Institut des Sciences de l'Evolution de Montpellier (UMR ISEM), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de recherche pour le développement [IRD] : UR226-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Research Institute of Freshwater Fisheries (CRIFI-RIFF), Research Institute for Aquaculture, Neovia Asia, Kasetsart University (KU), Universidade Federal de Santa Catarina = Federal University of Santa Catarina [Florianópolis] (UFSC), Embrapa Amazônia Ocidental, Partenaires INRAE, University of Oregon [Eugene], Swiss Institute of Bioinformatics [Lausanne] (SIB), Swiss Institute of Bioinformatics [Genève] (SIB), National Institutes of Health : R01 OD011116, National Institutes of Health: R35 GM139635, ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-16-CE12-0035,GenoFish,Evolution des génes et des génomes après duplication compléte(2016), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), MING WEN, State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Science, Hunan Normal University, QIAOWEI PAN, INRAE, LPGP, ELODIE JOUANNO, INRAE, LPGP, JEROME MONTFORT, INRAE, LPGP, MARGOT ZAHM, Plate-forme bio-informatique Genotoul, Mathématiques et Informatique Appliquées de Toulouse, INRAE, CÉDRIC CABAU, SIGENAE, GenPhySE, Université de Toulouse, INRAE, CHRISTOPHE KLOPP, SIGENAE, CAROLE IAMPIETRO, INRAE, CÉLINE ROQUES, INRAE, OLIVIER BOUCHEZ, INRAE, ADRIEN CASTINEL, INRAE, CÉCILE DONNADIEU, INRAE, HUGUES PARRINELLO, Montpellier GenomiX, CHARLES PONCET, GDEC Gentyane, INRAE, Université Clermont Auvergne, ELODIE BELMONTE, GDEC Gentyane, INRAE, Université Clermont Auvergne, VÉRONIQUE GAUTIER, GDEC Gentyane, INRAE, Université Clermont Auvergne, JEAN-CHRISTOPHE AVARRE, ISEM, CNRS, IRD, Univ Montpellier, REMI DUGUE, ISEM, CNRS, IRD, Univ Montpellier, RUDHY GUSTIANO, Research Institute of Freshwater Fisheries (CRIFI-RIFF), TRÂN THI THÚY HÀ, Research Institute for Aquaculture, MARC CAMPET, Neovia Asia, KEDNAPAT SRIPHAIROJ, Faculty of Natural Resources and Agro-Industry, JOSIANE RIBOLLI, Laboratório de Biologia e Cultivo de Peixes de Água Doce, Universidade Federal de Santa Catarina, FERNANDA LOUREIRO ALMEIDA OSULLIVAN, CPAA, THOMAS DESVIGNES, Institute of Neuroscience, University of Oregon, JOHN H. POSTLETHWAIT, Institute of Neuroscience, University of Oregon, CHRISTABEL FLOI BUCAO, Department of Ecology and Evolution, University of Lausanne, MARC ROBINSON-RECHAVI, Department of Ecology and Evolution, University of Lausanne, JULIEN BOBE, INRAE, LPGP, AMAURY HERPIN, INRAE, LPGP, and YANN GUIGUEN, INRAE, LPGP.
Subjects: Male, Male genome assembly, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Receptors, Peptide, Animals, Catfishes/genetics, Phylogeny, Receptors, Peptide/genetics, Receptors, Transforming Growth Factor beta/genetics, Y Chromosome/genetics, amhr2, evolution, male genome assembly, pangasiid catfishes, sex determination, Evolution, Pangasiid catfishes, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex determination, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, Peixe-gato, Y Chromosome, Genetics, Hormônio, Receptors, Transforming Growth Factor beta, Catfishes, Ecology, Evolution, Behavior and Systematics, Biotechnology
Abstract: The evolution of sex determination (SD) mechanisms in teleost fishes is amazingly dynamic, as reflected by the variety of different master sex-determining genes identified, even sometimes among closely related species. Pangasiids are a group of economically important catfishes in many South-Asian countries, but little is known about their sex determination system. Here, we generated novel genomic resources for 12 Pangasiid species and provided a first characterization of their SD system. Based on an Oxford Nanopore long-read chromosome-scale high quality genome assembly of the striped catfish Pangasianodon hypophthalmus, we identified a duplication of the anti-Müllerian hormone receptor type II gene (amhr2), which was further characterized as being sex-linked in males and expressed only in testicular samples. These first results point to a male-specific duplication on the Y chromosome (amhr2by) of the autosomal amhr2a. Sequence annotation revealed that the P. hypophthalmus Amhr2by is truncated in its N-terminal domain, lacking the cysteine-rich extracellular part of the receptor that is crucial for ligand binding, suggesting a potential route for its neofunctionalization. Short-read genome sequencing and reference-guided assembly of 11 additional Pangasiid species, along with sex-linkage studies, revealed that this truncated amhr2by duplication is also conserved as a male-specific gene in many Pangasiids. Reconstructions of the amhr2 phylogeny suggested that amhr2by arose from an ancient duplication / insertion event at the root of the Siluroidei radiation that is dated around 100 million years ago. Altogether these results bring multiple lines of evidence supporting that amhr2by is an ancient and conserved master sex-determining gene in Pangasiid catfishes, a finding that highlights the recurrent usage of the transforming growth factor β pathway in teleost sex determination and brings another empirical case towards the understanding of the dynamics or stability of sex determination systems.
Published: 2022

2. Generation of a chromosome‐level genome assembly for Pacific halibut ( Hippoglossus stenolepis ) and characterization of its sex‐determining genomic region

Author: Andrew J. Jasonowicz, Anna Simeon, Margot Zahm, Cédric Cabau, Christophe Klopp, Céline Roques, Carole Iampietro, Jérôme Lluch, Cécile Donnadieu, Hugues Parrinello, Daniel P. Drinan, Lorenz Hauser, Yann Guiguen, and Josep V. Planas
Subjects: Male, Fishes, Genetics, Animals, Female, Flounder, Genomics, Chromosomes, Ecosystem, Ecology, Evolution, Behavior and Systematics, Genome-Wide Association Study, Biotechnology
Abstract: The Pacific halibut (Hippoglossus stenolepis) is a key species in the North Pacific Ocean and Bering Sea ecosystems, where it also supports important fisheries. However, the lack of genomic resources limits our understanding of evolutionary, environmental and anthropogenic forces affecting key life history characteristics of Pacific halibut and prevents the application of genomic tools in fisheries management and conservation efforts. In the present study, we report on the first generation of a high-quality chromosome-level assembly of the Pacific halibut genome, with an estimated size of 602 Mb, 24 chromosome-length scaffolds that contain 99.8% of the assembly and a N
Published: 2022

3. Characterization of the sex determining region of channel catfish (Ictalurus punctatus) and development of a sex-genotyping test

Author: Ming Wen, Qiaowei Pan, Wes Larson, Camille Eché, and Yann Guiguen
Subjects: Male, Ictaluridae, Genome, Genotype, Genetic Linkage, Y Chromosome, Genetics, Animals, Female, General Medicine
Abstract: Channel catfish is an important species for aquaculture that exhibits a sexually dimorphic growth in favor of males. Genetic sexing and development of sex markers are crucial for the early identification of sex and of particular genotypes (YY males) for the production of all-male population in channel catfish aquaculture. In this study, we sequenced genomic DNA from pools of males and pools of females to better characterize the sex determining region (SDR) of channel catfish and to develop sex-specific markers for genetic sexing. Performing comparative analyses on male and female pooled genomic reads, we identified a large SDR (∼8.3 Mb) in the middle of channel catfish linkage group 4 (LG04). This non-recombining SDR contains a high-density of male-specific (Y chromosome) fixed single nucleotide polymorphisms (SNPs) along with ∼ 185 kb male-specific insertions or deletions. This SDR contains 95 annotated protein-encoding genes, including the recently reported putative channel catfish master sex determining (MSD) gene, breast cancer anti-estrogen resistance protein 1 (bcar1), located at one edge of the SDR. No sex-specific SNPs and/or indels were found in the coding sequence of bcar1, but one male-specific SNP was identified in its first intron. Based on this genomic information, we developed a PCR-based sex-specific genetic test. Genotyping results confirmed strong linkage between phenotypic sexes and the identified SDR in channel catfish. Our results confirm, using a Pool-Seq approach, that channel catfish is male heterogametic (XX-XY) with a large SDR on the LG04 sex chromosome. Furthermore, our genotyping primers can be used to identify XX, XY, and YY fish that will facilitate future research on sex determination and aquaculture applications in channel catfish.
Published: 2022

4. A supernumerary 'B-sex' chromosome drives male sex determination in the Pachón cavefish, Astyanax mexicanus

Author: Séverine Beille, Sylvie Rétaux, John H. Postlethwait, Claire Kuchly, Hugues Parrinello, Amaury Herpin, Tomáš Pavlica, Ahmed Al-Rikabi, Christophe Klopp, Cécile Donnadieu, Jorge Torres-Paz, Manfred Schartl, Lisa Gil, Thomas Liehr, Joerg Bohlen, Adrien Castinel, Romain Feron, Yann Guiguen, Céline Lopez-Roques, Margot Zahm, Cédric Cabau, Boudjema Imarazene, Frédéric Veyrunes, Thomas D. Mueller, Elodie Jouanno, Alexandr Sember, Kang Du, Sergey A. Simanovsky, Qiaowei Pan, Julie Perez, Laurent Journot, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Texas State University, Université de Lausanne = University of Lausanne (UNIL), Swiss Institute of Bioinformatics [Lausanne] (SIB), Génome et Transcriptome - Plateforme Génomique ( GeT-PlaGe), Plateforme Génome & Transcriptome (GET), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Génétique Physiologie et Systèmes d'Elevage (GenPhySE ), Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-École nationale supérieure agronomique de Toulouse (ENSAT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRAE), Institute of Animal Physiology and Genetics of the Czech Academy of Sciences (IAPG / CAS), Czech Academy of Sciences [Prague] (CAS), Charles University [Prague] (CU), Jena University Hospital [Jena], A.N. Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences [Moscow] (RAS), Institut des Sciences de l'Evolution de Montpellier (UMR ISEM), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Institut de recherche pour le développement [IRD] : UR226-Centre National de la Recherche Scientifique (CNRS), University of Würzburg = Universität Würzburg, University of Oregon [Eugene], This project was supported by funds from the 'Agence Nationale de la Recherche' (ANR/DFG, PhyloSex project, 2014-2016) to Y.G. and M.S. S.R. was supported by grants from an Equipe FRM (Fondation pour la Recherche Médicale, DEQ20150331745) and MITI CNRS (Mission pour les Initiatives Transverses et Interdisciplinaires). J.H.P. was supported by an NIH grant (R35 GM139635). Sequencing was supported by France Génomique as part of an 'Investissement d’avenir' program managed by ANR (contract ANR-10-INBS-09) and by the GET-PACBIO program (Programme opérationnel FEDER-FSE MIDI-PYRENEES ET GARONNE 2014-2020). The CytoEvol platform at ISEM was supported by the Labex CeMEB. J.B., T.P., and A.S. were supported by RVO: 67985904 of IAPG CAS, Liběchov. T.P. was supported by the projects of the Czech Ministry of Education (SVV 260571/2021). S.A.S. was supported by the Russian Foundation for Basic Research (RFBR) (18-34-00638). B.I.’s PhD fellowship was supported by the Doctoral School of Ecology, Geosciences, Agronomy, Nutrition of the University of Rennes 1 and INRAE. We are grateful to the genotoul bioinformatics platform Toulouse Occitanie (Bioinfo Genotoul, https://doi.org/10.15454/1.5572369328961167E12) for providing help, computing, and storage resources. Funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), University of Lausanne (UNIL), Université de Lausanne (UNIL), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-École nationale supérieure agronomique de Toulouse [ENSAT]-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRA), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche pour le développement [IRD] : UR226, Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École pratique des hautes études (EPHE), Guiguen, Yann, Blanc – Accords bilatéraux 2013 - Evolution des déterminants majeurs du sexe chez les poissons. - - PhyloSex2013 - ANR-13-ISV7-0005 - Blanc – Accords bilatéraux 2013 - VALID, and Organisation et montée en puissance d'une Infrastructure Nationale de Génomique - - France-Génomique2010 - ANR-10-INBS-0009 - INBS - VALID
Subjects: Male, [SDV]Life Sciences [q-bio], Male sex determination, sex determination, Cavefish, B chromosome, Biology, Genome, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, 0302 clinical medicine, Animals, gonads, Gene, genome, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Genetics, 0303 health sciences, Sexual differentiation, Characidae, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], sex chromosomes, Chromosome, Sex reversal, Biological Evolution, [SDV] Life Sciences [q-bio], Caves, cavefish, gdf6, sex differentiation, Female, General Agricultural and Biological Sciences, 030217 neurology & neurosurgery
Abstract: Sex chromosomes are generally derived from a pair of classical type-A chromosomes, and relatively few alternative models have been proposed up to now.(1,2) B chromosomes (Bs) are supernumerary and dispensable chromosomes with non-Mendelian inheritance found in many plant and animal species(3,4) that have often been considered as selfish genetic elements that behave as genome parasites.(5,6) The observation that in some species Bs can be either restricted or predominant in one sex(7–14) raised the interesting hypothesis that Bs could play a role in sex determination.(15) The characterization of putative B master sex-determining (MSD) genes, however, has not yet been provided to support this hypothesis. Here, in Astyanax mexicanus cavefish originating from Pachón cave, we show that Bs are strongly male predominant. Based on a high-quality genome assembly of a B-carrying male, we characterized the Pachón cavefish B sequence and found that it contains two duplicated loci of the putative MSD gene growth differentiation factor 6b (gdf6b). Supporting its role as an MSD gene, we found that the Pachón cavefish gdf6b gene is expressed specifically in differentiating male gonads, and that its knockout induces male-to-female sex reversal in B-carrying males. This demonstrates that gdf6b is necessary for triggering male sex determination in Pachón cavefish. Altogether these results bring multiple and independent lines of evidence supporting the conclusion that the Pachón cavefish B is a “B-sex” chromosome that contains duplicated copies of the gdf6b gene, which can promote male sex determination in this species.
Published: 2021

5. A brief review of vertebrate sex evolution with a pledge for integrative research: towards ‘ sexomics ’

Author: Yann Guiguen, Blanche Capel, Nicole Valenzuela, Qi Zhou, Michail Rovatsos, Manfred Schartl, Ben J. Evans, Matthias Stöck, Frédéric Veyrunes, Lukáš Kratochvíl, Tony Gamble, Alexander Suh, Heiner Kuhl, Institut des Sciences de l'Evolution de Montpellier (UMR ISEM), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre National de la Recherche Scientifique (CNRS)-Institut de recherche pour le développement [IRD] : UR226, Leibniz Institute of Freshwater Ecology and Inland Fisheries (IGB), Hiroshima University, Charles University [Prague] (CU), Department of Computing and Software (McMaster University), McMaster University [Hamilton, Ontario], University of East Anglia [Norwich] (UEA), Uppsala University, Iowa State University (ISU), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université de Montpellier (UM)-Institut de recherche pour le développement [IRD] : UR226-Centre National de la Recherche Scientifique (CNRS), Huzhou University [Zhejiang], University of Vienna [Vienna], Marquette University [Milwaukee], Duke University Medical Center, University of Würzburg, Texas State University, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), M.St. and M.Sc. were in part supported by COFASP/ERANET (STURGEoNOMICS) by the German Federal Ministry of Food and Agriculture through the Federal Office for Agriculture and Food (grant nos. 2816ERA04G, and 2816ERA05G), M.Sc. was also funded by the German Research Foundation (DFG), grant nos. SCHA408/14-1 und 15-1, H.K. was funded by the German Research Foundation (DFG), grant no. KU 3596/1-1 (project no. 324050651), L.K. and M.R. were supported by the Czech Science Foundation (project no. 17-22604S), B.J.E. was supported by the Natural Sciences and Engineering Research Council of Canada (RGPIN-2017-05770), A.S. was supported by the Swedish Research Council Vetenskapsrådet (grant nos. 2016-05139547 and 2020-04436), N.V. was supported by a grant from the National Science Foundation (grant no. IOS-1555999), B.C. was supported by a grant from the National Science Foundation (grant no. IOS-1256675), Q.Z. is supported by a European Research Council Starting Grant (grant no. 677696)., and Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-École pratique des hautes études (EPHE)
Subjects: Male, 0106 biological sciences, 0301 basic medicine, Sex Differentiation, media_common.quotation_subject, sex determination, Genomics, Fertility, [SDV.BID]Life Sciences [q-bio]/Biodiversity, 010603 evolutionary biology, 01 natural sciences, Genome, General Biochemistry, Genetics and Molecular Biology, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, Evolution, Molecular, Evolutionsbiologi, reproduction, 03 medical and health sciences, Genome Size, biology.animal, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Testis, evolution, genomics, Animals, Sex organ, Clade, Review Articles, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis, media_common, Evolutionary Biology, Dosage compensation, Sexual differentiation, biology, sex chromosomes, Ovary, Vertebrate, Articles, Sex Determination Processes, Biological Evolution, 030104 developmental biology, Evolutionary biology, Female, General Agricultural and Biological Sciences, vertebrates
Abstract: Triggers and biological processes controlling male or female gonadal differentiation vary in vertebrates, with sex determination (SD) governed by environmental factors or simple to complex genetic mechanisms that evolved repeatedly and independently in various groups. Here, we review sex evolution across major clades of vertebrates with information on SD, sexual development and reproductive modes. We offer an up-to-date review of divergence times, species diversity, genomic resources, genome size, occurrence and nature of polyploids, SD systems, sex chromosomes, SD genes, dosage compensation and sex-biased gene expression. Advances in sequencing technologies now enable us to study the evolution of SD at broader evolutionary scales, and we now hope to pursue asexomicsintegrative research initiative across vertebrates. The vertebratesexomecomprises interdisciplinary and integrated information on sexual differentiation, development and reproduction at all biological levels, from genomes, transcriptomes and proteomes, to the organs involved in sexual and sex-specific processes, including gonads, secondary sex organs and those with transcriptional sex-bias. Thesexomealso includes ontogenetic and behavioural aspects of sexual differentiation, including malfunction and impairment of SD, sexual differentiation and fertility. Starting from data generated by high-throughput approaches, we encourage others to contribute expertise to building understanding of thesexomesof many key vertebrate species.This article is part of the theme issue ‘Challenging the paradigm in sex chromosome evolution: empirical and theoretical insights with a focus on vertebrates (Part I)’.
Published: 2021

6. Allelic diversification after transposable element exaptation promoted gsdf as the master sex determining gene of sablefish

Author: Giles Goetz, Amaury Herpin, Alexandra Depincé, Yann Guiguen, Manfred Schartl, J. Adam Luckenbach, Edward S. Hayman, Krista M. Nichols, Aurélie Hua-Van, Anna Octavera, Julien Bobe, Goro Yoshizaki, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan University [Changsha] (HNU), Biocenter University of Würzburg = Biozentrum der Universität Würzburg, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), The Xiphophorus Genetic Stock Center, Department of Chemistry and Biochemistry, Texas State University, Evolution, génomes, comportement et écologie (EGCE), Institut de Recherche pour le Développement (IRD)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), NOAA National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric Administration (NOAA), Tokyo University of Marine Science and Technology (TUMSAT), University of Washington [Seattle], Center for Reproductive Biology, Washington State University (WSU), and ANR-16-COFA-0004,AquaCrispr,Optimization of the CRISPR/Cas9 knock-in technology and application in salmon and trout(2016)
Subjects: Male, Transposable element, [SDV]Life Sciences [q-bio], Locus (genetics), Biology, Y chromosome, Evolution, Molecular, 03 medical and health sciences, 0302 clinical medicine, Y Chromosome, Genetics, Animals, 14. Life underwater, Allele, Gene, Genetics (clinical), ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Research, Promoter, Genomics, Exaptation, Sex Determination Processes, Anoplopoma fimbria, Evolutionary biology, DNA Transposable Elements, 030217 neurology & neurosurgery, Heterogametic sex
Abstract: Concepts of evolutionary biology suggest that morphological change may occur by rare punctual but rather large changes, or by more steady and gradual transformations. It can therefore be asked whether genetic changes underlying morphological, physiological, and/or behavioral innovations during evolution occur in a punctual manner, whereby a single mutational event has prominent phenotypic consequences, or if many consecutive alterations in the DNA over longer time periods lead to phenotypic divergence. In the marine teleost, sablefish (Anoplopoma fimbria), complementary genomic and genetic studies led to the identification of a sex locus on the Y Chromosome. Further characterization of this locus resulted in identification of the transforming growth factor, beta receptor 1a (tgfbr1a) gene, gonadal somatic cell derived factor (gsdf), as the main candidate for fulfilling the master sex determining (MSD) function. The presence of different X and Y Chromosome copies of this gene indicated that the male heterogametic (XY) system of sex determination in sablefish arose by allelic diversification. The gsdfY gene has a spatio-temporal expression profile characteristic of a male MSD gene. We provide experimental evidence demonstrating a pivotal role of a transposable element (TE) for the divergent function of gsdfY. By insertion within the gsdfY promoter region, this TE generated allelic diversification by bringing cis-regulatory modules that led to transcriptional rewiring and thus creation of a new MSD gene. This points out, for the first time in the scenario of MSD gene evolution by allelic diversification, a single, punctual molecular event in the appearance of a new trigger for male development.
Published: 2021

7. A Y-linked anti-Müllerian hormone type-II receptor is the sex-determining gene in ayu, Plecoglossus altivelis

Author: Masatoshi Nakamoto, Ryusuke Sudo, Liu Wang, Yann Guiguen, Manfred Schartl, Eriko Koshimizu, Masato Endo, Takashi Sakamoto, Tsubasa Uchino, Ryota Sekiguchi, Yudai Kuchiishi, John H. Postlethwait, Tokyo University of Marine Science and Technology (TUMSAT), Yokohama City University (YCU), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Würzburg, Texas State University, University of Oregon [Eugene], This work was supported by funds from the Japan Society for the Promotion of Science (JSPS) KAKENHI (grant numbers 16K14972 and 20H00431 to TS and 18K05816 to MN), from the Japan International Cooperation Agency (JICA) for Science and Technology Research Partnership for Sustainable Development (SATREPS) to TS, from the 'Agence Nationale de la Recherche' and the 'Deutsche Forschungsgemeinschaft' (ANR/DFG, PhyloSex project, 2014-2016) to YG and MS, and from the USA National Institutes of Health to JHP (grant numbers R01OD011116 and 5R01GM085318)., and ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013)
Subjects: Male, Cancer Research, Single Nucleotide Polymorphisms, QH426-470, Genome, Ayu, Contig Mapping, 0302 clinical medicine, Loss of Function Mutation, Gene duplication, Medicine and Health Sciences, Genetics (clinical), Genetics, 0303 health sciences, Sex Characteristics, hormone anti-müllerienne, Genomics, Sex reversal, Genome wide association study of the useful traits in wild fish using the next generation sequencer, 次世代シーケンサーを用いた天然魚における優良形質の全ゲノム相関解析, Osmeriformes, Female, Anatomy, Plecoglossus altivelis, Genital Anatomy, Research Article, Fish Proteins, Receptors, Peptide, Biology, Y chromosome, Research and Analysis Methods, Synteny, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], 魚類における耐病性責任遺伝子の同定と新規疾病が野生集団に与える遺伝的影響の解析, Fish Genomics, Genome-Wide Association Studies, Plecoglossidae, Animals, 14. Life underwater, Gonads, Molecular Biology Techniques, Linkage Mapping, Molecular Biology, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, Sexual differentiation, Whole Genome Sequencing, Gene Mapping, Reproductive System, Biology and Life Sciences, Computational Biology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Human Genetics, biology.organism_classification, Genome Analysis, Animal Genomics, Y linkage, Receptors, Transforming Growth Factor beta, 030217 neurology & neurosurgery
Abstract: Whole-genome duplication and genome compaction are thought to have played important roles in teleost fish evolution. Ayu (or sweetfish), Plecoglossus altivelis, belongs to the superorder Stomiati, order Osmeriformes. Stomiati is phylogenetically classified as sister taxa of Neoteleostei. Thus, ayu holds an important position in the fish tree of life. Although ayu is economically important for the food industry and recreational fishing in Japan, few genomic resources are available for this species. To address this problem, we produced a draft genome sequence of ayu by whole-genome shotgun sequencing and constructed linkage maps using a genotyping-by-sequencing approach. Syntenic analyses of ayu and other teleost fish provided information about chromosomal rearrangements during the divergence of Stomiati, Protacanthopterygii and Neoteleostei. The size of the ayu genome indicates that genome compaction occurred after the divergence of the family Osmeridae. Ayu has an XX/XY sex-determination system for which we identified sex-associated loci by a genome-wide association study by genotyping-by-sequencing and whole-genome resequencing using wild populations. Genome-wide association mapping using wild ayu populations revealed three sex-linked scaffolds (total, 2.03 Mb). Comparison of whole-genome resequencing mapping coverage between males and females identified male-specific regions in sex-linked scaffolds. A duplicate copy of the anti-Müllerian hormone type-II receptor gene (amhr2bY) was found within these male-specific regions, distinct from the autosomal copy of amhr2. Expression of the Y-linked amhr2 gene was male-specific in sox9b-positive somatic cells surrounding germ cells in undifferentiated gonads, whereas autosomal amhr2 transcripts were detected in somatic cells in sexually undifferentiated gonads of both genetic males and females. Loss-of-function mutation for amhr2bY induced male to female sex reversal. Taken together with the known role of Amh and Amhr2 in sex differentiation, these results indicate that the paralog of amhr2 on the ayu Y chromosome determines genetic sex, and the male-specific amh-amhr2 pathway is critical for testicular differentiation in ayu., この研究のプレスリリース版はこちら: http://id.nii.ac.jp/1342/00002272/
Published: 2021

8. A nonfunctional copy of the salmonid sex-determining gene (sdY) is responsible for the 'apparent' XY females in Chinook salmon, Oncorhynchus tshawytscha

Author: Sylvain Bertho, Amaury Herpin, Elodie Jouanno, Ayaka Yano, Julien Bobe, Hugues Parrinello, Laurent Journot, René Guyomard, Thomas Muller, Penny Swanson, Garrett McKinney, Kevin Williamson, Mariah Meek, Manfred Schartl, Yann Guiguen, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Würzburg = Universität Würzburg, Physiological Chemistry, Biocenter, Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Julius-von-Sachs-Institute, Department of Molecular Plant Physiology and Biophysics, University of Würzburg, Environmental and Fisheries Sciences Division, Northwest Fisheries Science Center, National Oceanic and Atmospheric Administration (NOAA), Washington Department of Fish and Wildlife, Partenaires INRAE, Atreca, Inc., Michigan State University System, Texas State University, Scha408/12-1, 10-1, Deutsche Forschungsgemeinschaft, ANR-11-BSV7-0016,SDS,Rôle et évolution d'un possible déterminant majeur du sexe chez les Salmonidés.(2011), Guerineau, Nathalie C., and BLANC - Rôle et évolution d'un possible déterminant majeur du sexe chez les Salmonidés. - - SDS2011 - ANR-11-BSV7-0016 - BLANC - VALID
Subjects: sex reversal, Male, [SDV.BA] Life Sciences [q-bio]/Animal biology, [SDV]Life Sciences [q-bio], [SDV.BA]Life Sciences [q-bio]/Animal biology, sex determination, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex Determination Processes, [SDV] Life Sciences [q-bio], Salmon, salmonids, Testis, Genetics, Animals, Female, sdY, XY females, Gonads, Molecular Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, Genetics (clinical), Salmonidae
Abstract: Many salmonids have a male heterogametic (XX/XY) sex determination system, and they are supposed to have a conserved master sex-determining gene (sdY) that interacts at the protein level with Foxl2 leading to the blockage of the synergistic induction of Foxl2 and Nr5a1 of the cyp19a1a promoter. However, this hypothesis of a conserved master sex-determining role of sdY in salmonids is challenged by a few exceptions, one of them being the presence of naturally occurring “apparent” XY Chinook salmon, Oncorhynchus tshawytscha, females. Here, we show that some XY Chinook salmon females have a sdY gene (sdY-N183), with 1 missense mutation leading to a substitution of a conserved isoleucine to an asparagine (I183N). In contrast, Chinook salmon males have both a nonmutated sdY-I183 gene and the missense mutation sdY-N183 gene. The 3-dimensional model of SdY-I183N predicts that the I183N hydrophobic to hydrophilic amino acid change leads to a modification in the SdY β-sandwich structure. Using in vitro cell transfection assays, we found that SdY-I183N, like the wild-type SdY, is preferentially localized in the cytoplasm. However, compared to wild-type SdY, SdY-I183N is more prone to degradation, its nuclear translocation by Foxl2 is reduced, and SdY-I183N is unable to significantly repress the synergistic Foxl2/Nr5a1 induction of the cyp19a1a promoter. Altogether, our results suggest that the sdY-N183 gene of XY Chinook females is nonfunctional and that SdY-I183N is no longer able to promote testicular differentiation by impairing the synthesis of estrogens in the early differentiating gonads of wild Chinook salmon XY females.
Published: 2021

9. Genome biology of the darkedged splitfin

Author: Kang, Du, Martin, Pippel, Susanne, Kneitz, Romain, Feron, Irene, da Cruz, Sylke, Winkler, Brigitta, Wilde, Edgar G, Avila Luna, Gene, Myers, Yann, Guiguen, Constantino, Macias Garcia, and Manfred, Schartl
Subjects: Male, Cyprinodontiformes, Genome, Sex Chromosomes, Pregnancy, Animals, Female, Biology, Placentation
Abstract: Viviparity evolved independently about 150 times in vertebrates and more than 20 times in fish. Several lineages added to the protection of the embryo inside the body of the mother, the provisioning of nutrients, and physiological exchange. This often led to the evolution of a placenta. Among fish, one of the most complex systems serving the function of the placenta is the embryonal trophotaenia/ovarian luminal epithelium of the goodeid fishes. For a better understanding of this feature and others of this group of fishes, high-quality genomic resources are essential. We have sequenced the genome of the darkedged splitfin
Published: 2021

10. RADSex: A computational workflow to study sex determination using restriction site-associated DNA sequencing data

Author: Ørjan Karlsen, Susanne Kneitz, Rafael Henrique Nóbrega, Qiaowei Pan, Masatoshi Nakamoto, Elodie Jouanno, Thomas Desvignes, John H. Postlethwait, Ato Takanashi, Braedan M. McCluskey, Amaury Herpin, Matthias Stöck, Alvaro S. Roco, Boudjema Imarazene, Manfred Schartl, Zhongwei Wang, Mateus Contar Adolfi, Romain Feron, Catherine Wilson, Hugues Parrinello, Takashi Sakamoto, Ingo Braasch, Christophe Klopp, Marcos Antonio de Oliveira, Frederick W. Goetz, Harry William Detrich, Robert M. Waterhouse, Ming Wen, Yann Guiguen, Mari Kawaguchi, Angel Amores, Laurent Journot, Anna Wargelius, Verena A. Kottler, Kang Du, Jennifer L. Anderson, IINRAE, Univ Lausanne, Swiss Inst Bioinformat, Hunan Normal Univ, Uppsala Univ, Univ Montpellier, INRAE, Univ Wurzburg, Texas State Univ, Univ Oregon, NOAA, Sophia Univ, Northeastern Univ, Universidade Estadual Paulista (Unesp), Tokyo Univ Marine Sci & Technol, Inst Marine Res, Chinese Acad Sci, IGB, Michigan State Univ, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Department of Ecology and Evolution, University of Lausanne, Université de Lausanne (UNIL), Swiss Institute of Bioinformatics [Lausanne] (SIB), State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan University [Changsha] (HNU), Department of Systematic Biology [Uppsala], Evolutionary Biology Centre (EBC), Uppsala University-Uppsala University, Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Physiological Chemistry, Biocenter, University of Würzburg = Universität Würzburg, The Xiphophorus Genetic Stock Center, Department of Chemistry and Biochemistry, Texas State University, Institute of Neuroscience, University of Oregon, Eugene, University of Oregon [Eugene], Tokyo University of Marine Science and Technology (TUMSAT), Environmental and Fisheries Sciences Division, Northwest Fisheries Science Center, National Oceanic and Atmospheric Administration (NOAA), Sophia University, Tokyo, Faculty of Science and Technology, Department of Marine and Environmental Sciences, Marine Science Center, Northeastern University, Universidade Estadual Paulista Júlio de Mesquita Filho = São Paulo State University (UNESP), Institute of Marine Research [Bergen] (IMR), University of Bergen (UiB), Institute of Hydrobiology [Wuhan], Chinese Academy of Sciences [Beijing] (CAS), Leibniz Institute of Freshwater Ecology and Inland Fisheries (IGB), Department of Integrative Biology, Michigan State University, Michigan State University [East Lansing], Michigan State University System-Michigan State University System, This project was supported by funds from the 'Agence Nationale de la Recherche', the 'Deutsche Forschungsgemeinschaft' (ANR/DFG, PhyloSex project,2014-2016, SCHA 408/10-1, MS), the National Institutes of Health (USA) grant R01GM085318, JHP), and the National Science Foundation (USA) Office of Polar Programs (grants PLR-1247510 and PLR-1444167 to HWD and grant OPP-1543383 to JHP, TD, and HWD). The MGX core sequencing facility was supported by France Genomique National infrastructure, funded as part of 'Investissement d’avenir' program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09). RF was partially supported by Swiss National Science Foundation grant PP00P3_170664 to RMW., ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Department of Ecology and Evolution [Lausanne], Institute of Neuroscience, University of Oregon, Eugene, Oregon, Department of Materials and Life Sciences, Sophia University [Tokyo], Department of Structural and Functional Biology, Department of Aquatic Marine Biosciences, Developmental Biochemistry, Biocenter, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), and This project was supported by funds from the 'Agence Nationale de la Recherche', the 'Deutsche Forschungsgemeinschaft' (ANR/DFG, PhyloSex project, 2014-2016, SCHA 408/10-1, MS), the National Institutes of Health (USA) grants R01GM085318 and R35GM139635, JHP), and the National Science Foundation (USA) Office of Polar Programs (grants PLR-1247510 and PLR-1444167 to HWD and grant OPP-1543383 to JHP, TD, and HWD). The MGX core sequencing facility was supported by France Genomique National infrastructure, funded as part of 'Investissement d’avenir' program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09). RF was partially supported by Swiss National Science Foundation grant PP00P3_170664 to RMW.
Subjects: 0106 biological sciences, 0301 basic medicine, Male, Sex Determination Analysis, Computer science, [SDV]Life Sciences [q-bio], sex determination, Locus (genetics), Computational biology, Biology, Y chromosome, 010603 evolutionary biology, 01 natural sciences, Computational workflow, DNA sequencing, Article, Workflow, 03 medical and health sciences, Sex-determination system, Genetics, Animals, Sequencing, RAD&#8208, Computational Biology, DNA, Female, Fishes/genetics, Sequence Analysis, DNA, Sex Chromosomes, Software, RAD-Sequencing, computational workflow, fish, visualization, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, Visualization, 0303 health sciences, Fishes, Japanese Medaka, Sex determination, Restriction site, 030104 developmental biology, Fish, Identification (biology), Biotechnology
Abstract: Made available in DSpace on 2021-06-25T11:54:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-09 National Institute of Health (USA) Deutsche Forschungsgemeinschaft National Science Foundation (USA) Office of Polar Programs Agence Nationale de la Recherche Schweizerischer Nationalfonds zur Forderung der Wissenschaftlichen Forschung The study of sex determination and sex chromosome organization in nonmodel species has long been technically challenging, but new sequencing methodologies now enable precise and high-throughput identification of sex-specific genomic sequences. In particular, restriction site-associated DNA sequencing (RAD-Seq) is being extensively applied to explore sex determination systems in many plant and animal species. However, software specifically designed to search for and visualize sex-biased markers using RAD-Seq data is lacking. Here, we present RADSex, a computational analysis workflow designed to study the genetic basis of sex determination using RAD-Seq data. RADSex is simple to use, requires few computational resources, makes no prior assumptions about the type of sex-determination system or structure of the sex locus, and offers convenient visualization through a dedicated R package. To demonstrate the functionality of RADSex, we re-analysed a published data set of Japanese medaka, Oryzias latipes, where we uncovered a previously unknown Y chromosome polymorphism. We then used RADSex to analyse new RAD-Seq data sets from 15 fish species spanning multiple taxonomic orders. We identified the sex determination system and sex-specific markers in six of these species, five of which had no known sex-markers prior to this study. We show that RADSex greatly facilitates the study of sex determination systems in nonmodel species thanks to its speed of analyses, low resource usage, ease of application and visualization options. Furthermore, our analysis of new data sets from 15 species provides new insights on sex determination in fish. IINRAE, LPGP, Rennes, France Univ Lausanne, Dept Ecol & Evolut, Lausanne, Switzerland Swiss Inst Bioinformat, Lausanne, Switzerland Hunan Normal Univ, Coll Life Sci, State Key Lab Dev Biol Freshwater Fish, Changsha, Peoples R China Uppsala Univ, Dept Organismal Biol, Systemat Biol, Uppsala, Sweden Univ Montpellier, Inst Genom Fonct, INSERM, IGF,CNRS, Montpellier, France INRAE, Math & Informat Appl Toulouse, SIGENAE, Castanet Tolosan, France Univ Wurzburg, Physiol Chem, Bioctr, Wurzburg, Germany Texas State Univ, Xiphophorus Genet Stock Ctr, Dept Chem & Biochem, San Marcos, TX USA Univ Wurzburg, Dev Biochem, Bioctr, Wurzburg, Germany Univ Oregon, Inst Neurosci, Eugene, OR USA NOAA, Environm & Fisheries Sci Div, Northwest Fisheries Sci Ctr, Natl Marine Fisheries Serv, Seattle, WA USA Sophia Univ, Fac Sci & Technol, Dept Mat & Life Sci, Tokyo, Japan Northeastern Univ, Ctr Marine Sci, Dept Marine & Environm Sci, Nahant, MA 01908 USA Sao Paulo State Univ, Inst Biosci, Dept Struct & Funct Biol, Reprod & Mol Biol Grp, Botucatu, SP, Brazil Tokyo Univ Marine Sci & Technol, Dept Aquat Marine Biosci, Tokyo, Japan Inst Marine Res, Bergen, Norway Chinese Acad Sci, Inst Hydrobiol, Beijing, Peoples R China IGB, Leibniz Inst Freshwater Ecol & Inland Fishe, Berlin, Germany Michigan State Univ, Dept Integrat Biol, Ecol Evolut & Behav Program, E Lansing, MI 48824 USA Sao Paulo State Univ, Inst Biosci, Dept Struct & Funct Biol, Reprod & Mol Biol Grp, Botucatu, SP, Brazil National Institute of Health (USA): R01GM085318 National Institute of Health (USA): R35GM139635 Deutsche Forschungsgemeinschaft: SCHA 408/10-1 National Science Foundation (USA) Office of Polar Programs: OPP-1543383 National Science Foundation (USA) Office of Polar Programs: PLR-1247510 National Science Foundation (USA) Office of Polar Programs: PLR-1444167 Agence Nationale de la Recherche: SCHA 408/10-1 Agence Nationale de la Recherche: ANR-10-INBS-09 Schweizerischer Nationalfonds zur Forderung der Wissenschaftlichen Forschung: PP00P3_170664
Published: 2021

11. Genetic determinism of spontaneous masculinisation in XX female rainbow trout: new insights using medium throughput genotyping and whole-genome sequencing

Author: Charles Poncet, Florent Petitprez, Maria Bernard, Edwige Quillet, Chris Hoze, Nicolas Dechamp, Philippe Hocdé, Anastasia Bestin, Céline Ciobotaru, Yann Guiguen, Pierrick Haffray, Marine Milhes, Florence Phocas, Olivier Bouchez, Jérôme Lluch, Clémence Fraslin, Mathieu Charles, Francine Krieg, Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Edinburgh, Syndicat des Sélectionneurs Avicoles et Aquacoles Français (SYSAAF), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Allice, Ligue Nationale Contre le Cancer - Paris, Ligue Nationale Contre le Cancer (LNCC), Génome et Transcriptome - Plateforme Génomique ( GeT-PlaGe), Plateforme Génome & Transcriptome (GET), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génétique Diversité et Ecophysiologie des Céréales (GDEC), Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Pisciculture Charles Murgat, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), European Maritime and Fisheries Fund and France Agrimer co-funded this work (NeoBio project, n° RFEA470016FA1000008), Ligue Nationnale Contre le Cancer, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse)
Subjects: 0106 biological sciences, 0301 basic medicine, Male, Candidate gene, QTL, animal diseases, [SDV]Life Sciences [q-bio], Masculinisation des poissons, Genome-wide association study, heritability, 01 natural sciences, Genome-wide association studies, Genetics, 0303 health sciences, education.field_of_study, Multidisciplinary, biology, Reproduction, 04 agricultural and veterinary sciences, Genomics, Poisson, Trout, Phenotype, Oncorhynchus mykiss, maleness, Medicine, Female, Sex, Maleness, Heterogametic sex, endocrine system, animal structures, Genotype, Science, Population, Quantitative trait locus, Déterminisme génétique du sexe, 010603 evolutionary biology, Article, Heritability, 03 medical and health sciences, Salmonide, Genetic variation, sex, Animals, 14. Life underwater, education, 030304 developmental biology, Genetic association, Animal breeding, fish, Whole Genome Sequencing, urogenital system, Truite arc-en-ciel, 0402 animal and dairy science, Sex Determination Processes, biology.organism_classification, 040201 dairy & animal science, 030104 developmental biology, Fish, Rainbow trout, Sequençage
Abstract: Rainbow trout has a male heterogametic (XY) sex determination system controlled by a major sex-determining gene, sdY. Unexpectedly, a few phenotypically masculinised fish are regularly observed in all-female farmed trout stocks. To better understand the genetic determinism underlying spontaneous maleness in XX-rainbow trout, we recorded the phenotypic sex of 20,210 XX-rainbow trout from a French farm population at 10 and 15 months post-hatching. The overall masculinisation rate was 1.45%. We performed two genome-wide association studies (GWAS) on a subsample of 1139 individuals classified as females, intersex or males using either medium-throughput genotyping (31,811 SNPs) or whole-genome sequencing (WGS, 8.7 million SNPs). The genomic heritability of maleness ranged between 0.48 and 0.62 depending on the method and the number of SNPs used for the estimation. At the 31K SNPs level, we detected four QTL on three chromosomes (Omy1, Omy12 and Omy20). Using WGS information, we narrowed down the positions of the two QTL detected on Omy1 to 96 kb and 347 kb respectively, with the second QTL explaining up to 14% of the total genetic variance of maleness. Within this QTL, we detected three putative candidate genes, fgfa8, cyp17a1 and an uncharacterised protein (LOC110527930), which might be involved in spontaneous maleness of XX-female rainbow trout.
Published: 2020

12. Genome Sequence of the Euryhaline Javafish Medaka

Author: Yusuke, Takehana, Margot, Zahm, Cédric, Cabau, Christophe, Klopp, Céline, Roques, Olivier, Bouchez, Cécile, Donnadieu, Celia, Barrachina, Laurent, Journot, Mari, Kawaguchi, Shigeki, Yasumasu, Satoshi, Ansai, Kiyoshi, Naruse, Koji, Inoue, Chuya, Shinzato, Manfred, Schartl, Yann, Guiguen, and Amaury, Herpin
Subjects: Male, Salinity, whole genome sequencing, Genome, Oryzias, adaptation, Adaptation, Physiological, Medaka, Genome Report, Osmoregulation, long reads, Models, Animal, evolution, Animals, Female, genetic map, transcriptome, Phylogeny
Abstract: The genus Oryzias consists of 35 medaka-fish species each exhibiting various ecological, morphological and physiological peculiarities and adaptations. Beyond of being a comprehensive phylogenetic group for studying intra-genus evolution of several traits like sex determination, behavior, morphology or adaptation through comparative genomic approaches, all medaka species share many advantages of experimental model organisms including small size and short generation time, transparent embryos and genome editing tools for reverse and forward genetic studies. The Java medaka, Oryzias javanicus, is one of the two species of medaka perfectly adapted for living in brackish/sea-waters. Being an important component of the mangrove ecosystem, O. javanicus is also used as a valuable marine test-fish for ecotoxicology studies. Here, we sequenced and assembled the whole genome of O. javanicus, and anticipate this resource will be catalytic for a wide range of comparative genomic, phylogenetic and functional studies. Complementary sequencing approaches including long-read technology and data integration with a genetic map allowed the final assembly of 908 Mbp of the O. javanicus genome. Further analyses estimate that the O. javanicus genome contains 33% of repeat sequences and has a heterozygosity of 0.96%. The achieved draft assembly contains 525 scaffolds with a total length of 809.7 Mbp, a N50 of 6,3 Mbp and a L50 of 37 scaffolds. We identified 21454 predicted transcripts for a total transcriptome size of 57, 146, 583 bps. We provide here a high-quality chromosome scale draft genome assembly of the euryhaline Javafish medaka (321 scaffolds anchored on 24 chromosomes (representing 97.7% of the total bases)), and give emphasis on the evolutionary adaptation to salinity.
Published: 2020

13. Independent origin of XY and ZW sex determination mechanisms in Mosquitofish sister species

Author: Susanne Kneitz, Frederik Helmprobst, Hugues Parrinello, Céline Lopez-Roques, Kang Du, Indrajit Nanda, Romain Feron, Verena A. Kottler, Dunja K. Lamatsch, Christophe Klopp, Laurent Journot, Manfred Schartl, Jérôme Lluch, Yann Guiguen, University of Würzburg = Universität Würzburg, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institute of Bioinformatics, GSF - Research Institute for Environment and Health, Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRA), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), University of Innsbruck, Génome et Transcriptome - Plateforme Génomique (GeT-PlaGe), Institut National de la Recherche Agronomique (INRA)-Plateforme Génome & Transcriptome (GET), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Montpellier (UM), Hagler Institute for Advanced Study and Department of Biology, Texas A&M University System, This project was supported by funds to Y.G. and M.S. from the 'Agence Nationale de la Recherche' and the 'Deutsche Forschungsgemeinschaft' (ANR/DFG, PhyloSex project, 2014-2016). V.A.K. was supported through a SCIENTIA fellowship of the Free State of Bavaria. The GeT, Toulouse (C.L.-R., J.L.), and the Montpellier GenomiX (MGX) France (L.J., H.P.) core facilities were supported by France Génomique National infrastructure, funded as part of 'Investissement d’avenir' program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09)., ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3)
Subjects: Male, 0106 biological sciences, X Chromosome, Genetic Linkage, 46, XX Testicular Disorders of Sex Development, [SDV]Life Sciences [q-bio], sex determination, Locus (genetics), sox9b, Investigations, Y chromosome, 010603 evolutionary biology, 01 natural sciences, Gambusia, Cyprinodontiformes, 03 medical and health sciences, Y Chromosome, Genetics, Animals, gipc1, Cell Lineage, [INFO]Computer Science [cs], Allele, Phylogeny, 030304 developmental biology, 0303 health sciences, [SDV.GEN]Life Sciences [q-bio]/Genetics, Genome, Sex Chromosomes, Autosome, biology, Mosquitofish, Pigmentation, Chromosome Mapping, Chromosome, Eastern mosquitofish, Sex Determination Processes, biology.organism_classification, W chromosome, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, Phenotype, Female, pool sequencing
Abstract: Fish are known for the outstanding variety of their sex determination mechanisms and sex chromosome systems. The western (Gambusia affinis) and eastern mosquitofish (G. holbrooki) are sister species for which different sex determination mechanisms have been described: ZZ/ZW for G. affinis and XX/XY for G. holbrooki. Here, we carried out restriction-site associated DNA (RAD-) and pool sequencing (Pool-seq) to characterize the sex chromosomes of both species. We found that the ZW chromosomes of G. affinis females and the XY chromosomes of G. holbrooki males correspond to different linkage groups, and thus evolved independently from separate autosomes. In interspecific hybrids, the Y chromosome is dominant over the W chromosome, and X is dominant over Z. In G. holbrooki, we identified a candidate region for the Y-linked melanic pigmentation locus, a rare male phenotype that constitutes a potentially sexually antagonistic trait and is associated with other such characteristics, e.g., large body size and aggressive behavior. We developed a SNP-based marker in the Y-linked allele of GIPC PDZ domain containing family member 1 (gipc1), which was linked to melanism in all tested G. holbrooki populations. This locus represents an example for a color locus that is located in close proximity to a putative sex determiner, and most likely substantially contributed to the evolution of the Y.
Published: 2020

14. Sex chromosome and sex locus characterization in goldfish, Carassius auratus (Linnaeus, 1758)

Author: Ming, Wen, Romain, Feron, Qiaowei, Pan, Justine, Guguin, Elodie, Jouanno, Amaury, Herpin, Christophe, Klopp, Cedric, Cabau, Margot, Zahm, Hugues, Parrinello, Laurent, Journot, Shawn M, Burgess, Yoshihiro, Omori, John H, Postlethwait, Manfred, Schartl, Yann, Guiguen, Hunan Normal University, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), University of Lausanne, Lausanne, Switzerland, Swiss Institute of Bioinformatics [Lausanne] (SIB), Université de Lausanne (UNIL), Plateforme Bio-Informatique - Génotoul, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Institut de Génomique Fonctionnelle - Montpellier GenomiX (IGF MGX), Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), National Human Genome Research Institute (NHGRI), Nagahama Institute of Bioscience and Technology, Osaka University [Osaka], University of Oregon [Eugene], University of Würzburg, Texas State University, Grant-in-Aid for Scientific Research (19 K22426) - Grants: R01OD011116 and 5R01GM085318 from the USA National Institutes of Health - ANAEE-France National infrastructure, ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-10- INBS0009,INBS,France-Génomique, Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan University [Changsha] (HNU), University of Lausanne (UNIL), Département d'écologie et évolution [Lausanne] (DEE), Génétique Physiologie et Systèmes d'Elevage (GenPhySE ), Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-École nationale supérieure agronomique de Toulouse [ENSAT]-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRA), Centre National de la Recherche Scientifique (CNRS)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1)-Centre National de la Recherche Scientifique (CNRS)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1), Institute for Protein Research [Osaka], Nagahama Institute of Bio-Science and Technology, Oregon State University (OSU), 'Agence Nationale de la Recherche' and the 'Deutsche Forschungsgemeinschaft' (ANR/DFG,PhyloSex project, 2014–2016) to MS and YG. Montpellier Genomics (MGX)facility was supported by France Génomique National infrastructure, fundedas part of 'Investissement d’avenir' program managed by Agence Nationalepour la Recherche (contract ANR-10-INBS-09). This work was also supportedby Grant-in-Aid for Scientific Research (19 K22426) to YO, and grantsR01OD011116 and 5R01GM085318 from the USA National Institutes of Healthto JHP. INRAE-U3E was supported by a PEARL INRAE 1036 U3E funding fromthe ANAEE-France National infrastructure., ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université de Lausanne = University of Lausanne (UNIL), Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRAE), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-BioCampus (BCM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Naudinat, Carine, Organisation et montée en puissance d'une Infrastructure Nationale de Génomique - - France-Génomique2010 - ANR-10-INBS-0009 - INBS - VALID, and Blanc – Accords bilatéraux 2013 - Evolution des déterminants majeurs du sexe chez les poissons. - - PhyloSex2013 - ANR-13-ISV7-0005 - Blanc – Accords bilatéraux 2013 - VALID
Subjects: Male, Male genome assembly, [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], Sex Chromosomes, lcsh:QH426-470, Sex markers, Genetic Linkage, [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT], lcsh:Biotechnology, [SDV]Life Sciences [q-bio], Poolseq, RADseq, Sex Determination Processes, Sex determination, [SDV] Life Sciences [q-bio], lcsh:Genetics, Y Chromosome, lcsh:TP248.13-248.65, Goldfish, Animals, Female, Goldfish/genetics, Sex Chromosomes/genetics, Sex Determination Processes/genetics, Research Article
Abstract: International audience; Background - Goldfish is an important model for various areas of research, including neural development and behavior and a species of significant importance in aquaculture, especially as an ornamental species. It has a male heterogametic (XX/XY) sex determination system that relies on both genetic and environmental factors, with high temperatures being able to produce female-to-male sex reversal. Little, however, is currently known on the molecular basis of genetic sex determination in this important cyprinid model. Here we used sequencing approaches to better characterize sex determination and sex-chromosomes in an experimental strain of goldfish. Results - Our results confirmed that sex determination in goldfish is a mix of environmental and genetic factors and that its sex determination system is male heterogametic (XX/XY). Using reduced representation (RAD-seq) and whole genome (pool-seq) approaches, we characterized sex-linked polymorphisms and developed male specific genetic markers. These male specific markers were used to distinguish sex-reversed XX neomales from XY males and to demonstrate that XX female-to-male sex reversal could even occur at a relatively low rearing temperature (18 °C), for which sex reversal has been previously shown to be close to zero. We also characterized a relatively large non-recombining region (~ 11.7 Mb) on goldfish linkage group 22 (LG22) that contained a high-density of male-biased genetic polymorphisms. This large LG22 region harbors 373 genes, including a single candidate as a potential master sex gene, i.e., the anti-Mullerian hormone gene (amh). However, no sex-linked polymorphisms were detected in the coding DNA sequence of the goldfish amh gene. Conclusions - These results show that our goldfish strain has a relatively large sex locus on LG22, which is likely the Y chromosome of this experimental population. The presence of a few XX males even at low temperature also suggests that other environmental factors in addition to temperature could trigger female-to-male sex reversal. Finally, we also developed sex-linked genetic markers, which will be important tools for future research on sex determination in our experimental goldfish population. However, additional work would be needed to explore whether this sex locus is conserved in other populations of goldfish.
Published: 2020

15. The rise and fall of the ancient northern pike master sex-determining gene

Author: Mike Tringali, Hongxia Hu, Amaury Herpin, Patrick W. DeHaan, Eric Rondeau, Louis Bernatchez, Wesley A. Larson, Manfred Schartl, Elodie Jouanno, E. A. Interesova, Yann Guiguen, Eric Saillant, Stephen S. Curran, Hugo Verreycken, Gaël P.J. Denys, Roger A. Tabor, Vladimir A. Trifonov, Hugo Darras, Christophe Klopp, Hugues Parrinello, Frederick W. Goetz, John H. Postlethwait, Qiaowei Pan, Céline Roques, J. Andrés López, Camille Eché, Jérôme Lluch, Laurent Journot, Frank A. von Hippel, Krista M. Nichols, Konrad Ocalewicz, Romain Feron, René Guyomard, Song-Lin Chen, Ben F. Koop, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Department of Ecology and Evolution [Lausanne], Université de Lausanne (UNIL), Swiss Institute of Bioinformatics [Lausanne] (SIB), Department of Biology, University of Victoria, Northwest Fisheries Science Center (NWFSC), NOAA National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric Administration (NOAA)-National Oceanic and Atmospheric Administration (NOAA), Alaska Pacific University, Institut de Biologie Intégrative et des Systèmes [Québec] (IBIS), Florida Marine Research Institute, Florida Fish and Wildlife Conservation Commission, School of Ocean Science and Technology [Mississippi] (SOST), University of Southern Mississippi (USM), Gulf Coast Research Laboratory, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Patrimoine naturel (PatriNat), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS)-Office français de la biodiversité (OFB), Department of Biological Sciences [Flagstaff], Northern Arizona University [Flagstaff], Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, College of Fisheries and Ocean Sciences (CFOS), University of Alaska [Fairbanks] (UAF), Research Institute for Nature and Forest (INBO), Department of Marine Biology and Ecology, University of Gdansk, Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Beijing Fisheries Research Institute, U.S. Fish and Wildlife Service, Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Tomsk State University [Tomsk], Siberian Branch of the Russian Academy of Sciences (SB RAS), Developmental Biochemistry, Biocenter, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Institute of Neuroscience, University of Oregon, Eugene, Oregon, University of Oregon [Eugene], This work was supported by the Agence Nationale de la Recherche, the Deutsche Forschungsgemeinschaft (ANR/DFG, PhyloSex project, 2014–2016, SCHA 408/10–1, to YG and MS), and the U.S. National Institutes of Health (R01GM085318 to JHP). The MGX and Get-Plage core sequencing facilities were supported by France Genomique National infrastructure, funded as part of the ‘Investissement d’avenir’ program managed by the Agence Nationale pour la Recherche (contract ANR-10-INBS-09)., ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Université de Lausanne = University of Lausanne (UNIL), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université Toulouse III - Paul Sabatier (UT3), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), University of Victoria [Canada] (UVIC), Qingdao National Laboratory for Marine Science and Technology, University of Gdańsk (UG), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Texas State University, Agence Nationale de la Recherche, Deutsche Forschungsgemeinschaft (ANR/DFG, PhyloSex project, 2014–2016, SCHA 408/10–1, to YG and MS), U.S. National Institutes of Health (R01GM085318 to JHP), France Genomique National infrastructure, funded as part of the ‘Investissement d’avenir’ program managed by the Agence Nationale pour la Recherche (contract ANR-10-INBS-09), Julius-Maximilians-Universität Würzburg (JMU), Corvaisier, Maryse, Blanc – Accords bilatéraux 2013 - Evolution des déterminants majeurs du sexe chez les poissons. - - PhyloSex2013 - ANR-13-ISV7-0005 - Blanc – Accords bilatéraux 2013 - VALID, Organisation et montée en puissance d'une Infrastructure Nationale de Génomique - - France-Génomique2010 - ANR-10-INBS-0009 - INBS - VALID, and Przeworski, Molly (ed.)
Subjects: Male, 0106 biological sciences, 0301 basic medicine, Evolution of sexual reproduction, Lineage (evolution), [SDV]Life Sciences [q-bio], sex determination, 01 natural sciences, mudminnows, Gene Duplication, Gene duplication, Biology (General), Clade, master sex determining gene, Phylogeny, северная щука, computer.programming_language, гены, определяющие пол, 0303 health sciences, education.field_of_study, Sex Chromosomes, biology, umbridae, General Neuroscience, General Medicine, [SDV] Life Sciences [q-bio], Medicine, Female, Research Article, QH301-705.5, Demographic history, Science, Population, esocidae, Y chromosome, 010603 evolutionary biology, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Animals, education, Esox, pikes, 030304 developmental biology, Pike, fish, General Immunology and Microbiology, evolutionary biology, Sex Determination Processes, biology.organism_classification, esociforms, Sexual reproduction, 030104 developmental biology, Evolutionary biology, Other, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, computer, Sex linkage
Abstract: Sexual reproduction is a ubiquitous basic feature of life and genetic sex determination is thus widespread, at least among eukaryotes. Understanding the remarkable diversity of sex determination mechanisms, however, is limited by the paucity of empirical studies. Here, we traced back the evolution of sex determination in an entire clade of vertebrates and uncovered that the northern pike (Esox lucius) master sex-determining gene initiated from a 65 to 90 million-year-old gene duplication and remained sex-linked on undifferentiated sex chromosomes for at least 56 million years. Contrasting with its ancient origin, we identified several independent species- or population-specific transitions of sex determination mechanisms in this lineage, including an unexpected complete and recent Y-chromosome loss in some North American northern pike populations. These findings highlight the diversity of the evolutionary fates of master sex-determining genes and raise the importance of careful considerations of population demographic history in sex determination studies. Our study also puts forward the hypothesis that occasional sex reversals and genetic bottlenecks provide a non-adaptive explanation for sex determination transitions.
Published: 2020

16. GWAS reveals several genomic regions governing spontaneous XX-maleness in rainbow trout

Author: Clémence Fraslin, Anastasia Bestin, Nicolas Dechamp, Ambrosio, Jonathan D., Francine Krieg, Elodie Belmonte, Charles Poncet, Philippe Hocdé, Pierrick Haffray, Yann Guiguen, Florence Phocas, Edwige Quillet, Génétique Animale et Biologie Intégrative (GABI), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, INRA Laboratoire de Physiologie et Génomique des Poissons, Syndicats des sélectionneurs avicoles et aquacoles français (SYSAAF), Génétique Diversité et Ecophysiologie des Céréales (GDEC), Institut National de la Recherche Agronomique (INRA)-Université Blaise Pascal - Clermont-Ferrand 2 (UBP), Pisciculture Charles Murgat, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), The European Maritime and Fisheries Fund and FranceAgrimer funded this work (NeoBio project, n° R FEA470016FA1000008), Syndicat des Sélectionneurs Avicoles et Aquacoles Français (SYSAAF), Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), European Aquaculture Society (EAS). BEL., and Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de la Recherche Agronomique (INRA)
Subjects: fish, salmonidae, endocrine system, trout, détermination du sexe, animal structures, urogenital system, animal diseases, [SDV]Life Sciences [q-bio], gène déterminant majeur du sexe, sex determination, digestive system, gonad, génétique de la variance, gonade, reproduction, mâle, poisson, rainbow, salmonids, température, ComputingMilieux_MISCELLANEOUS, truite arc en ciel, expression des gènes
Abstract: GWAS reveals several genomic regions governing spontaneous XX-maleness in rainbow trout. Aquaculture Europe 2019
Published: 2019

17. Identification of the master sex determining gene in Northern pike (Esox lucius) reveals restricted sex chromosome differentiation

Author: Amaury Herpin, Ming Wen, Jean-Paul Concordet, Julien Bobe, Jean-Mickael Busnel, René Guyomard, Estelle Vigouroux, Ayaka Yano, Jérôme Lluch, Laurent Journot, Elodie Jouanno, Romain Feron, Manfred Schartl, Yann Guiguen, Christophe Klopp, Qiaowei Pan, Hugues Parrinello, John H. Postlethwait, Céline Roques, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), Department of Ecology and Evolution [Lausanne], Université de Lausanne (UNIL), Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Fédération d'Ille-et-Vilaine pour la pêche et la protection du milieu aquatique [Rennes] (FDPPMA 35), Structure et Instabilité des Génomes (STRING), Muséum national d'Histoire naturelle (MNHN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Plateforme Bio-Informatique - Génotoul, Institut National de la Recherche Agronomique (INRA), Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Génétique Physiologie et Systèmes d'Elevage (GenPhySE ), École nationale supérieure agronomique de Toulouse [ENSAT]-Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Génome et Transcriptome - Plateforme Génomique (GeT-PlaGe), Institut National de la Recherche Agronomique (INRA)-Plateforme Génome & Transcriptome (GET), Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institute of Neuroscience [Eugene, OR, États-Unis], University of Oregon [Eugene], Department of Physiological Chemistry [Würzburg, Allemagne], University of Würzburg-Biocenter [Würzburg, Allemagne], Comprehensive Cancer Center Mainfranken [Würzburg, Allemagne], University Hospital of Würzburg, Department of Biology [College Station, TX, États-Unis], Hagler Institute for Advanced Study [College Station, TX, États-Unis], Texas A&M University [College Station]-Texas A&M University [College Station], This project was supported by funds from the 'Agence Nationale de la Recherche' and the 'Deutsche Forschungsgemeinschaft' (ANR/DFG, PhyloSex project, 2014-2016) to YG and MS. The GeT core facility (JL, CR), Toulouse, France and the MGX facility, Montpellier, France (LJ, HP) were supported by France Génomique National infrastructure, funded as part of 'Investissement d’avenir' program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09)., ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Department of Ecology and Evolution, Sorbonne Université (SU), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Unité de Mathématiques et Informatique Appliquées de Toulouse (MIAT INRA), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Institute of Neuroscience, Physiological Chemistry, Biocenter, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Comprehensive Cancer Center Mainfranken, University Clinic Würzburg, Hagler Institute for Advanced Study and Department of Biology, Texas A&M University System, This project was supported by funds from the 'Agence Nationale de la Recherche' and the 'Deutsche Forschungsgemeinschaft' (ANR/ DFG, PhyloSex project, 2014-2016) to YG and MS. The GeT core facility (JL, CR), Toulouse, France and the MGX facility, Montpellier, France (LJ, HP) were supported by France Génomique National infrastructure, funded as part of 'Investissement d’avenir' program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09), Bodescot, Myriam, Blanc – Accords bilatéraux 2013 - Evolution des déterminants majeurs du sexe chez les poissons. - - PhyloSex2013 - ANR-13-ISV7-0005 - Blanc – Accords bilatéraux 2013 - VALID, Organisation et montée en puissance d'une Infrastructure Nationale de Génomique - - France-Génomique2010 - ANR-10-INBS-0009 - INBS - VALID, Université de Lausanne = University of Lausanne (UNIL), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-École nationale supérieure agronomique de Toulouse [ENSAT], Dpt of Ecology and Evolution [Lausanne], Muséum national d'Histoire naturelle (MNHN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), BioCampus Montpellier (BCM), Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Plateforme Bioinformatique Genotoul [Castanet-Tolosan], Institut National de la Recherche Agronomique (INRA)-Institut National de la Recherche Agronomique (INRA), GeT PlaGe, Genotoul, ANR-10-INBS-09-01/10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Julius-Maximilians-Universität Würzburg (JMU), and ProdInra, Migration
Subjects: Anti-Mullerian Hormone, Male, Evolution of sexual reproduction, [SDV]Life Sciences [q-bio], gène déterminant majeur du sexe, [MATH] Mathematics [math], [SDV.GEN] Life Sciences [q-bio]/Genetics, QH426-470, Animals, Genetically Modified, mâle, Database and Informatics Methods, 0302 clinical medicine, duplication de gènes, poisson, Gene Duplication, Gene duplication, Morphogenesis, Medicine and Health Sciences, chromosome sexuel, [MATH]Mathematics [math], Phylogeny, Genetics, détermination du sexe, 0303 health sciences, education.field_of_study, Sex Chromosomes, Sexual Differentiation, Chromosome Biology, Chromosome Mapping, Animals, Anti-Mullerian Hormone/genetics, Esocidae/physiology, Female, Gene Knockdown Techniques, Sex Chromosomes/genetics, Sex Determination Processes/genetics, Synteny, Genomics, [SDV] Life Sciences [q-bio], séquençage du génome, hormone antimullérienne, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Anatomy, Sequence Analysis, Genital Anatomy, différenciation sexuelle, expression des gènes, Research Article, Bioinformatics, Population, Locus (genetics), [SDV.GEN.GA] Life Sciences [q-bio]/Genetics/Animal genetics, [INFO] Computer Science [cs], Biology, Research and Analysis Methods, DNA sequencing, Chromosomes, reproduction, 03 medical and health sciences, esox lucius, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [INFO]Computer Science [cs], education, Molecular Biology Techniques, Gonads, Gene, Molecular Biology, 030304 developmental biology, Whole genome sequencing, [SDV.GEN]Life Sciences [q-bio]/Genetics, Sequence Assembly Tools, génome, Gene Mapping, Reproductive System, Chromosome, Biology and Life Sciences, Computational Biology, Cell Biology, Sex Determination, Sex Determination Processes, Genome Analysis, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, Genetic Loci, Esocidae, Sequence Alignment, 030217 neurology & neurosurgery, Developmental Biology
Abstract: Teleost fishes, thanks to their rapid evolution of sex determination mechanisms, provide remarkable opportunities to study the formation of sex chromosomes and the mechanisms driving the birth of new master sex determining (MSD) genes. However, the evolutionary interplay between the sex chromosomes and the MSD genes they harbor is rather unexplored. We characterized a male-specific duplicate of the anti-Müllerian hormone (amh) as the MSD gene in Northern Pike (Esox lucius), using genomic and expression evidence as well as by loss-of-function and gain-of-function experiments. Using RAD-Sequencing from a family panel, we identified Linkage Group (LG) 24 as the sex chromosome and positioned the sex locus in its sub-telomeric region. Furthermore, we demonstrated that this MSD originated from an ancient duplication of the autosomal amh gene, which was subsequently translocated to LG24. Using sex-specific pooled genome sequencing and a new male genome sequence assembled using Nanopore long reads, we also characterized the differentiation of the X and Y chromosomes, revealing a small male-specific insertion containing the MSD gene and a limited region with reduced recombination. Our study reveals an unexpectedly low level of differentiation between a pair of sex chromosomes harboring an old MSD gene in a wild teleost fish population, and highlights both the pivotal role of genes from the amh pathway in sex determination, as well as the importance of gene duplication as a mechanism driving the turnover of sex chromosomes in this clade., Author summary In stark contrast to mammals and birds, a high proportion of teleosts have homomorphic sex chromosomes and display a high diversity of sex determining genes. Yet, population level knowledge of both the sex chromosome and the master sex determining gene is only available for the Japanese medaka, a model species. Here we identified and provided functional proofs of an old duplicate of anti-Müllerian hormone (Amh), a member of the Tgf- β family, as the male master sex determining gene in the Northern pike, Esox lucius. We found that this duplicate, named amhby (Y-chromosome-specific anti-Müllerian hormone paralog b), was translocated to the sub-telomeric region of the new sex chromosome, and now amhby shows strong sequence divergence as well as substantial expression pattern differences from its autosomal paralog, amha. We assembled a male genome sequence using Nanopore long reads and identified a restricted region of differentiation within the sex chromosome pair in a wild population. Our results provide insight on the conserved players in sex determination pathways, the mechanisms of sex chromosome turnover, and the diversity of levels of differentiation between homomorphic sex chromosomes in teleosts.
Published: 2019

18. Transcriptome of tambaqui Colossoma macropomum during gonad differentiation: Different molecular signals leading to sex identity

Author: Dany Severac, Vanessa Ribeiro Reis, Fernanda Loureiro de Almeida, Yann Guiguen, Gilvan Ferreira da Silva, Aldessandro da Costa Amaral, Áquila Rodrigues do Nascimento, Igor Kelvyn Cavalcante Lobo, Michel Eduardo Beleza Yamagishi, Embrapa Amazônia Ocidental, Embrapa Informática Agropecuária, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Génomique Fonctionnelle - Montpellier GenomiX (IGF MGX), Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Universidade Federal do Amazonas, The study wasfinancially supported by Embrapa (SEG03.13.09.010.00.00), the fellowship grants were supported by ADAPTA-II (465540/2014-7) to Igor Kelvyn Cavalcante Lobo and by CAPES to Vanessa Ribeiro Reis, France Génomique National infrastructure funded as part of Investissement d'avenir 'program managed by Agence Nationale pour la Recherche (contract ANR-10-INBS-09), and ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010)
Subjects: 0106 biological sciences, Male, Gonad, Sex Differentiation, Tambaqui, [SDV]Life Sciences [q-bio], Population, Serrasalmide, 01 natural sciences, Mâle, Transcriptome, 03 medical and health sciences, Testis, Genetics, medicine, Juvenile, Expression des gènes, Animals, 14. Life underwater, education, Gonadal Steroid Hormones, Gene, Colossoma macropomum, Wnt Signaling Pathway, 030304 developmental biology, Reproduction animale, 0303 health sciences, education.field_of_study, Sexual differentiation, biology, Wnt/beta-catenin pathway, Détermination du sexe, Ovary, Wnt signaling pathway, Femelle, biology.organism_classification, Différenciation sexuelle, Poisson, Biosynthetic Pathways, medicine.anatomical_structure, Female, Sequençage, Characiformes, 010606 plant biology & botany
Abstract: International audience; Tambaqui (Colossoma macropomum) is the major native species in Brazilian aquaculture, and we have shown that females exhibit a higher growth compared to males, opening up the possibility for the production of all-female population. To date, there is no information on the sex determination and differentiation molecular mechanisms of tambaqui. In the present study, transcriptome sequencing of juvenile trunks was performed to understand the molecular network involved in the gonadal sex differentiation. The results showed that before differentiation, components of the Wnt/beta-catenin pathway, fox and fst genes imprint female sex development, whereas antagonistic pathways (gsk3b, wt1 and fgfr2), sox9 and genes for androgen synthesis indicate male differentiation. Hence, in undifferentiated tambaqui, the Wnt/beta-catenin exerts a role on sex differentiation, either upregulated in female-like individuals, or antagonized in male-like individuals.
Published: 2019

19. The unusual rainbow trout sex determination gene hijacked the canonical vertebrate gonadal differentiation pathway

Author: Eric Pailhoux, Barbara Nicol, Maëlle Pannetier, Misako Miwa, Amaury Herpin, Manfred Schartl, Yann Guiguen, Adèle Branthonne, Elodie Jouanno, Ayaka Yano, Sylvain Bertho, Goro Yoshizaki, Thomas Müller, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Physiological Chemistry, Biocenter, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences [Durham] (NIEHS-NIH), National Institutes of Health [Bethesda] (NIH)-National Institutes of Health [Bethesda] (NIH), Julius-von-Sachs-Institute, Department of Molecular Plant Physiology and Biophysics, University of Würzburg, Biologie du développement et reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Department of Marine Biosciences, Tokyo University of Marine Science and Technology (TUMSAT), University Hospital, Comprehensive Cancer Center Mainfranken, Hagler Institute for Advanced Study, Texas A&M University [College Station], Department of Biology, University of Minho, This work was supported by Agence Nationale de la Recherche (ANR) Grant ANR-11-BSV7- 0016 (SDS project) and grants from the Deutsche Forschungsgemeinschaft (Scha408/12-1, 10-1, to M.S.), Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and ANR-11-BSV7-0016,SDS,Rôle et évolution d'un possible déterminant majeur du sexe chez les Salmonidés.(2011)
Subjects: 0301 basic medicine, Forkhead Box Protein L2, Male, réseau de régulation de géne, Sex Differentiation, [SDV]Life Sciences [q-bio], gène déterminant majeur du sexe, sex determination, sexual differentiation, Translocation, Genetic, 0302 clinical medicine, poisson, Transcription (biology), salmonids, Gene duplication, Testis, Gene Regulatory Networks, Aromatase, Promoter Regions, Genetic, différenciation gonadique, Genetics, salmonidae, trout, détermination du sexe, Multidisciplinary, Vertebrate, Cell Differentiation, Biological Sciences, gène sdY, Oncorhynchus mykiss, Female, transcription, différenciation sexuelle, expression des gènes, Biology, reproduction, 03 medical and health sciences, Forkhead Transcription Factors, rainbow, biology.animal, evolution, Animals, Gonads, Gene, Transcription factor, fish, Cell Nucleus, Sexual differentiation, Forkhead box proteins, immunisation génétique, Estrogens, Sex Determination Processes, vertébré, 030104 developmental biology, biology.protein, vertebrates, 030217 neurology & neurosurgery, truite arc en ciel
Abstract: Evolutionary novelties require rewiring of transcriptional networks and/or the evolution of new gene functions. Sex determination (SD), one of the most plastic evolutionary processes, requires such novelties. Studies on the evolution of vertebrate SD revealed that new master SD genes are generally recruited from genes involved in the downstream SD regulatory genetic network. Only a single exception to this rule is currently known in vertebrates: the intriguing case of the salmonid master SD gene (sdY), which arose from duplication of an immune-related gene. This exception immediately posed the question of how a gene outside from the classical sex differentiation cascade could acquire its function as a male SD gene. Here we show that SdY became integrated in the classical vertebrate sex differentiation cascade by interacting with the Forkhead box domain of the female-determining transcription factor, Foxl2. In the presence of Foxl2, SdY is translocated to the nucleus where the SdY:Foxl2 complex prevents activation of the aromatase (cyp19a1a) promoter in cooperation with Nr5a1 (Sf1). Hence, by blocking a positive loop of regulation needed for the synthesis of estrogens in the early differentiating gonad, SdY disrupts a preset female differentiation pathway, consequently allowing testicular differentiation to proceed. These results also suggest that the evolution of unusual vertebrate master sex determination genes recruited from outside the classical pathway like sdY is strongly constrained by their ability to interact with the canonical gonadal differentiation pathway.
Published: 2018

20. Evolution of sex hormone binding globulins reveals early gene duplication at the root of vertebrates

Author: Julien Bobe, Yann Guiguen, Jérémy Pasquier, Alexis Fostier, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and ANR-10-GENM-0017,PhyloFish,Analyse phylogénomique des duplications géniques chez les poissons téléostéens: une approche par RNA-Seq(2010)
Subjects: 0106 biological sciences, Male, Teleost, [SDV]Life Sciences [q-bio], 01 natural sciences, Transcriptome, 0302 clinical medicine, Endocrinology, Sex hormone-binding globulin, Gene Duplication, Sex Hormone-Binding Globulin, Hormone sexuelle, Gene expression, Gene duplication, Expression des gènes, Bowfin, Phylogeny, Genetics, 0303 health sciences, biology, Reproduction, Vertebrate, Poisson, Spotted gar, Génomique, Vertebrates, Female, Sex hormone-binding globulin SHBG, Evolution, 030209 endocrinology & metabolism, teleosts, 010603 evolutionary biology, Synteny, Evolution, Molecular, 03 medical and health sciences, Protein Domains, biology.animal, Animals, Humans, Amino Acid Sequence, Gonads, Gene, 030304 developmental biology, Gene Expression Profiling, Lamprey, biology.organism_classification, Gene Expression Regulation, gene expression, biology.protein, Animal Science and Zoology, Duplication de gènes
Abstract: Sex hormone-binding globulin (Shbg) is an important vertebrate blood carrier protein synthetized in the liver and involved in the transport and local regulation of sex steroids in target tissues. A novel shbg gene (shbgb) with a predominant ovarian expression was recently characterized. Being initially found only in salmonids, this shbgb was originally thought to result from the Salmonid-specific whole genome duplication. Using updated transcriptomic and genomic resources we identified Shbgb orthologs in non-salmonid teleosts (European eel, arowana), holosteans (spotted gar, bowfin), polypteriformes (reedfish), agnatha (sea lamprey) and in amphibians, and found that the classical Shbg gene (Shbga) displays a predominant hepatic expression whereas Shbgb has a predominant gonadal expression. Together, these results indicate that these two Shgb genes most likely originate from a whole genome duplication event at the root of vertebrate evolution, followed by numerous and independent losses and by tissue expression specialization of Shbga and Shbgb paralogs.HighlightsPhylogeny, synteny and expression analyses shed new light on Shbg evolution in vertebrates.Shbg diversity originates from a duplication event at the root of vertebrate evolution.This duplication was followed by many independent losses of Shbg paralogs in vertebrates.Shbg paralogs have acquired different tissue expression patterns.
Published: 2021

21. Foxl2 and Its Relatives Are Evolutionary Conserved Players in Gonadal Sex Differentiation

Author: Qiaowei Pan, Yann Guiguen, Jeremy Pasquier, Manfred Schartl, Eric Pailhoux, Sylvain Bertho, Amaury Herpin, Julien Bobe, Gaël Le Trionnaire, John H. Postlethwait, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Physiological Chemistry, Biocenter, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Comprehensive Cancer Center Mainfranken, University Hospital of Würzburg, Biologie des Organismes et Ecosystèmes Aquatiques (BOREA), Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Institut de Génétique, Environnement et Protection des Plantes (IGEPP), Institut National de la Recherche Agronomique (INRA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-AGROCAMPUS OUEST, Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Institute of Neuroscience, University of Oregon [Eugene], Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Texas Institute for Advanced Study and Department of Biology, Texas A&M University System, This work was supported by grants from the ‘Agence Nationale de la Recherche’ (ANR) ANR-10-GENM-017 (Phylo- Fish project), ANR ANR-11-BSV7-0016 (SDS project) and by a joined funding from ANR and the Deutsche Forschungsgemeinschaft (ANR/DFG, PhyloSex project, 2014–2016). Sylvain Bertho was supported by an INRA/Region PhD fellowship followed by the ‘DAAD STIBET Doktoranden’ Program., ANR-10-GENM-0017,PhyloFish,Analyse phylogénomique des duplications géniques chez les poissons téléostéens: une approche par RNA-Seq(2010), ANR-11-BSV7-0016,SDS,Rôle et évolution d'un possible déterminant majeur du sexe chez les Salmonidés.(2011), ANR-13-ISV7-0005,PhyloSex,Evolution des déterminants majeurs du sexe chez les poissons.(2013), Muséum national d'Histoire naturelle (MNHN), Sorbonne Universités, Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Muséum national d'Histoire naturelle (MNHN)-Institut de Recherche pour le Développement (IRD)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université des Antilles (UA), Institut de Recherche pour le Développement (IRD [Nouvelle-Calédonie]), Université Pierre et Marie Curie - Paris 6 (UPMC), Normandie Université (NU), Institut National de la Recherche Agronomique (INRA), Sorbonne Université (SU)-Université des Antilles (UA)-Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS)-Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut de Recherche pour le Développement (IRD), Julius-Maximilians-Universität Würzburg (JMU), Institut National de la Recherche Agronomique (INRA)-Université de Rennes (UR)-AGROCAMPUS OUEST, and École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)
Subjects: Forkhead Box Protein L2, Male, 0301 basic medicine, Embryology, Gonad, Somatic cell, [SDV]Life Sciences [q-bio], Endocrinology, Diabetes and Metabolism, Biology, Evolution, Molecular, 03 medical and health sciences, foxl3, Forkhead Transcription Factors, foxl2, evolution, Gene duplication, medicine, Animals, Humans, Gonads, Gene, Phylogeny, Genetics, Sexual differentiation, Germ Cells, 030104 developmental biology, Forkhead box L2, medicine.anatomical_structure, Vertebrates, sex differentiation, fox genes, Female, Germ cell, Developmental Biology
Abstract: Foxl2 is a member of the large family of Forkhead Box (Fox) domain transcription factors. It emerged during the last 15 years as a key player in ovarian differentiation and oogenesis in vertebrates and especially mammals. This review focuses on Foxl2 genes in light of recent findings on their evolution, expression, and implication in sex differentiation in animals in general. Homologs of Foxl2 and its paralog Foxl3 are found in all metazoans, but their gene evolution is complex, with multiple gains and losses following successive whole genome duplication events in vertebrates. This review aims to decipher the evolutionary forces that drove Foxl2/3 gene specialization through sub- and neo-functionalization during evolution. Expression data in metazoans suggests that Foxl2/3 progressively acquired a role in both somatic and germ cell gonad differentiation and that a certain degree of sub-functionalization occurred after its duplication in vertebrates. This generated a scenario where Foxl2 is predominantly expressed in ovarian somatic cells and Foxl3 in male germ cells. To support this hypothesis, we provide original results showing that in the pea aphid (insects) foxl2/3 is predominantly expressed in sexual females and showing that in bovine ovaries FOXL2 is specifically expressed in granulosa cells. Overall, current results suggest that Foxl2 and Foxl3 are evolutionarily conserved players involved in somatic and germinal differentiation of gonadal sex.
Published: 2016

22. Dynamic and differential expression of the gonadal aromatase during the process of sexual differentiation in a novel transgenic cyp19a1a -eGFP zebrafish line

Author: Jean-Marc Porcher, Nathalie Hinfray, Edith Chadili, Morgane Caulier, Yann Guiguen, Benjamin Piccini, Frédéric Sohm, Camille Torchy, François Brion, Institut National de l'Environnement Industriel et des Risques (INERIS), Plateforme d'ingénierie génétique des animaux modèles (AMAGEN), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Agence Nationale de la Recherche, Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), This work was supported by the French National Ministry of Ecology and Sustainable Development [Program 190 ECOT AP 2013-2016, NEMO project and Program 181-DRC 50] and by the French National Research Program for Environmental and Occupational Health of Anses [AIDEZ project n° EST-2016/1/189]. AMAGEN received the support of the French National Research Agency (ANR) 'Investissement d’Avenir' program [TEFOR ANR-II-INSB-0014]., and ANR-11-INBS-0014,TEFOR,Transgenèse pour les Etudes Fonctionnelles sur les Organismes modèles(2011)
Subjects: Male, 0301 basic medicine, green fluorescent protein, Embryo, Nonmammalian, Sex Differentiation, [SDV]Life Sciences [q-bio], sexual differentiation, poisson zèbre, Green fluorescent protein, gonade, Animals, Genetically Modified, Endocrinology, poisson, cyprinidae, Testis, poisson transgenique, Aromatase, cyp19a1a, Zebrafish, protéine fluorescente verte, endocrine disruptor, perturbateur endocrinien, Gene Expression Regulation, Developmental, gonadal aromatase, Cell biology, medicine.anatomical_structure, danio rerio, lignée transgénique, Female, Development of the gonads, danio devario, différenciation sexuelle, expression des gènes, aromatase, medicine.medical_specialty, Gonad, Transgene, Green Fluorescent Proteins, Biology, gonad, Gene Expression Regulation, Enzymologic, reproduction, 03 medical and health sciences, Internal medicine, medicine, Animals, 14. Life underwater, transgenic zebrafish, Gonads, fish, Sexual differentiation, fungi, ovaire, Zebrafish Proteins, biology.organism_classification, Sexual dimorphism, Germ Cells, 030104 developmental biology, biology.protein, Animal Science and Zoology, ovary
Abstract: Accepted manuscript; In zebrafish, there exists a clear need for new tools to study sex differentiation dynamic and its perturbation by endocrine disrupting chemicals. In this context, we developed and characterized a novel transgenic zebrafish line expressing green fluorescent protein (GFP) under the control of the zebrafish cyp19a1a (gonadal aromatase) promoter. In most gonochoristic fish species including zebrafish, cyp19a1a, the enzyme responsible for the synthesis of estrogens, has been shown to play a critical role in the processes of reproduction and sexual differentiation. This novel cyp19a1a-eGFP transgenic line allowed a deeper characterization of expression and localization of cyp19a1a gene in zebrafish gonads both at the adult stage and during development. At the adult stage, GFP expression was higher in ovaries than in testis. We showed a perfect co-expression of GFP and endogenous Cyp19a1a protein in gonads that was mainly localized in the cytoplasm of peri-follicular cells in the ovary and of Leydig and germ cells in the testis. During development, GFP was expressed in all immature gonads of 20 dpf-old zebrafish. Then, GFP expression increased in early differentiated female at 30 and 35 dpf to reach a high GFP intensity in well-differentiated ovaries at 40 dpf. On the contrary, males consistently displayed low GFP expression as compared to female whatever their stage of development, resulting in a clear dimorphic expression between both sexes. Interestingly, fish that undergoes ovary-to-testis transition (35 and 40 dpf) presented GFP levels similar to males or intermediate between females and males. In this transgenic line our results confirm that cyp19a1a is expressed early during development, before the histological differentiation of the gonads, and that the down-regulation of cyp19a1a expression is likely responsible for the testicular differentiation. Moreover, we show that although cyp19a1a expression exhibits a clear dimorphic expression pattern in gonads during sexual differentiation, its expression persists whatever the sex suggesting that estradiol synthesis is important for gonadal development of both sexes. Monitoring the expression of GFP in control and exposed-fish will help determine the sensitivity of this transgenic line to EDCs and to refine mechanistic based-assays for the study of EDCs. In fine, this transgenic zebrafish line will be a useful tool to study physiological processes such as reproduction and sexual differentiation, and their perturbations by EDCs.
Published: 2018

23. Differential expression of subunits of 20β-hydroxysteroid dehydrogenase during gametogenesis in rainbow trout (Oncorhychus mykiss)

Author: Yann Guiguen, Arya Vazirzadeh, Laboratoire de Physiologie et Génomique des Poissons (LPGP), and Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )
Subjects: 0301 basic medicine, Male, gamétogenèse, [SDV]Life Sciences [q-bio], gonade, Endocrinology, Food Animals, poisson, salmonids, gonadogénèse, Hydroxysteroid dehydrogenase, salmonidae, trout, oncorhynchus mykiss, biology, General Medicine, rainbow trout, different tissues, Trout, Female, expression des gènes, medicine.medical_specialty, endocrine system, animal structures, Protein subunit, gonad, Gene Expression Regulation, Enzymologic, gametogenesis, Andrology, reproduction, 03 medical and health sciences, rainbow, Internal medicine, expression, medicine, Animals, Gametogenesis, fish, Head Kidney, urogenital system, 20β-HSD subunits, Hydroxysteroid Dehydrogenases, biology.organism_classification, Protein Subunits, 030104 developmental biology, cellule somatique, Animal Science and Zoology, Rainbow trout, somatic cell, Vitellogenesis, Hormone, truite arc en ciel
Abstract: The patterns of expression of two subunits of 20β-hydroxysteroid dehydrogenase (20β-HSD), key enzyme involved in the biosynthesis and activation of steroid hormones, were examined in rainbow trout by using a combination of quantitative real-time PCR and in-situ hybridization. The expression of targeted genes was examined in mRNA extracted from different tissues at different gonadal stages in male and female trout. Both subunits of 20β-HSD were found to be widely distributed in tissues. The highest expression of 20β-HSD A was found in intestine followed by skin, stomach, liver and gills, whereas, the highest expression of 20β-HSD B was observed in stomach followed by head kidney, ovary − at late vitellogenesis stage- and trunk kidney. In ovarian tissue 20β-HSD A was highly expressed in mature oocytes, and the highest expression of 20β-HSD B was in ovary at late vitellogenesis stage. There were no differences in the level of expression of either subunit among groups of rainbow trout at different stages of maturational competence. In male fish, 20β-HSD A was highly expressed in testis stage I in contrast to 20β-HSD B which was highly expressed in testis stage VIII. In situ- hybridization results showed that the 20β-HSD gene was highly expressed in gastrointestinal organs, while only slightly expressed in the gonadal tissue of fish at stage 62 day-post-fertilization (dpf). Overall, the results confirm the ubiquitous presence of 20β-HSD among tissues in rainbow trout with relatively minor fluctuations in expression associated with reproductive cycles which collectively suggests a wider metabolic role of these enzymes than just an association with the synthesis of control hormones for reproduction.
Published: 2017

24. Heritable Targeted Inactivation of the Rainbow Trout (Oncorhynchus mykiss) Master Sex-Determining Gene Using Zinc-Finger Nucleases

Author: Yann Guiguen, Elodie Jouanno, Barbara Nicol, Ayaka Yano, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), ANR SVSE 7-2011, Project SDS, Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), and European Project: 222719,EC:FP7:KBBE,FP7-KBBE-2007-2A,LIFECYCLE(2009)
Subjects: Male, endocrine system, Molecular Sequence Data, Biology, medicine.disease_cause, Applied Microbiology and Biotechnology, gene knockout, Frameshift mutation, 03 medical and health sciences, Transforming Growth Factor beta, medicine, Animals, sdY, 14. Life underwater, Genes, sry, Gene, Gene knockout, 030304 developmental biology, Genetics, Sex Characteristics, 0303 health sciences, Mutation, Base Sequence, [SDV.BA]Life Sciences [q-bio]/Animal biology, Gene targeting, Zinc Fingers, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, 04 agricultural and veterinary sciences, Sex Determination Processes, Zebrafish Proteins, Sex reversal, Endonucleases, rainbow trout, Stop codon, Oncorhynchus mykiss, Gene Targeting, 040102 fisheries, 0401 agriculture, forestry, and fisheries, Female, Rainbow trout, Genetic Engineering, zfn
Abstract: Gene targeting is a powerful tool for analyzing gene function. Recently, new technology for gene targeting using engineered zinc-finger nucleases (ZFNs) has been described in fish species. However, it has not yet been widely used for cold water and slow developing species, such as Salmonidae. Here, we present the results of successful ZFN-mediated disruption of the sex-determining gene sdY (sexually dimorphic on the Y chromosome) in rainbow trout (Oncorhynchus mykiss). Three pairs of ZFN mRNA targeted to different regions of the sdY gene were injected into fertilized rainbow trout eggs. Sperm from 1-year-old male founders (parental generation one or P1) carrying a ZFN-induced mutation in their germline were then used to produce F1 non-mosaic animals. In these F1 populations, we characterized 14 different mutations in the sdY gene, including one mutation leading to the deletion of leucine 43 (L43) and 13 mutations at other target sites that had different effects on the SdY protein, i.e., amino acid insertions, deletions, and frameshift mutations producing premature stop codons in the mRNA. The gonadal phenotype analysis of the F1-mutated animals revealed that the single L43 amino acid deletion did not lead to a male-to-female sex reversal, but all other mutations induced a clear ovarian phenotype. These results show that targeted gene disruption using ZFN is efficient in rainbow trout but depends on the ZFN design. We also characterized new sdY mutations resulting in male-to-female sex reversal, and we conclude that L43 seems dispensable for SdY function.
Published: 2013

25. Divergent Expression Regulation of Gonad Development Genes in Medaka Shows Incomplete Conservation of the Downstream Regulatory Network of Vertebrate Sex Determination

Author: Manfred Schartl, Amaury Herpin, Mareen Engel, Yann Guiguen, Johanna Klughammer, Minoru Tanaka, Maria Hinzmann, Cornelia Schmidt, Barbara Nicol, Mateus Contar Adolfi, Physiological Chemistry, Biocenter, Am Hubland, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Laboratory of Molecular Genetics for Reproduction, National Institute for Basic Biology [Okazaki], DFG (GK 1048, Molecular basis of organ development in vertebrates) and 'Mechanisms of sex determination in zebrafish' (UO Subaward Number 212791A), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), European Project: 222719,EC:FP7:KBBE,FP7-KBBE-2007-2A,LIFECYCLE(2009), and Julius-Maximilians-Universität Würzburg (JMU)
Subjects: Fish Proteins, Male, gonadal development, [SDV]Life Sciences [q-bio], Oryzias, Gene regulatory network, Biology, Cell Line, Animals, Genetically Modified, Evolution, Molecular, 03 medical and health sciences, Aromatase, 0302 clinical medicine, Gene expression, Genetics, Transcriptional regulation, Animals, Gene Regulatory Networks, ddc:610, Gonads, Wnt Signaling Pathway, divergent expression regulation, Molecular Biology, Hedgehog, Gene, Transcription factor, Discoveries, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, Regulation of gene expression, 0303 health sciences, Binding Sites, Granulosa Cells, adaptive evolution, Wnt signaling pathway, Gene Expression Regulation, Developmental, Membrane Proteins, Forkhead Transcription Factors, Sex Determination Processes, Theca Cells, Female, 030217 neurology & neurosurgery, Transcription Factors, gene regulatory network evolution
Abstract: Genetic control of male or female gonad development displays between different groups of organisms a remarkable diversity of “master sex-determining genes” at the top of the genetic hierarchies, whereas downstream components surprisingly appear to be evolutionarily more conserved. Without much further studies, conservation of sequence has been equalized to conservation of function. We have used the medaka fish to investigate the generality of this paradigm. In medaka, the master male sex-determining gene is dmrt1bY, a highly conserved downstream regulator of sex determination in vertebrates. To understand its function in orchestrating the complex gene regulatory network, we have identified targets genes and regulated pathways of Dmrt1bY. Monitoring gene expression and interactions by transgenic fluorescent reporter fish lines, in vivo tissue-chromatin immunoprecipitation and in vitro gene regulation assays revealed concordance but also major discrepancies between mammals and medaka, notably amongst spatial, temporal expression patterns and regulations of the canonical Hedgehog and R-spondin/Wnt/Follistatin signaling pathways. Examination of Foxl2 protein distribution in the medaka ovary defined a new subpopulation of theca cells, where ovarian-type aromatase transcriptional regulation appears to be independent of Foxl2. In summary, these data show that the regulation of the downstream regulatory network of sex determination is less conserved than previously thought.
Published: 2013

26. Expression profiling of candidate genes during ovary-to-testis trans-differentiation in rainbow trout masculinized by androgens

Author: Alexis Fostier, Rémi Houlgatte, Yann Guiguen, Daniel Baron, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), Physiopathologie et pharmacologie cellulaires et moléculaires, Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM), and Université de Nantes (UN)
Subjects: Male, Sex Differentiation, Cellular differentiation, Oogenesis, 0302 clinical medicine, Endocrinology, Testis, sex-inversion, gonads, Oligonucleotide Array Sequence Analysis, 0303 health sciences, education.field_of_study, Reverse Transcriptase Polymerase Chain Reaction, Leydig Cells, Cell Differentiation, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, fishes, Sertoli cell, rainbow trout, medicine.anatomical_structure, Data Interpretation, Statistical, Multigene Family, Oncorhynchus mykiss, Female, Genetic Markers, endocrine system, medicine.medical_specialty, medicine.drug_class, Granulosa cell, salmonid, Population, Biology, gametogenesis, Andrology, 03 medical and health sciences, Internal medicine, medicine, Animals, education, DNA Primers, 030304 developmental biology, Sexual differentiation, urogenital system, Gene Expression Profiling, Ovary, androgens, Androgen, Gene expression profiling, Sex steroid, Cell Transdifferentiation, sex-differentiation, Animal Science and Zoology, 030217 neurology & neurosurgery
Abstract: International audience; Fish gonadal phenotype is very sensitive to sex steroid and functional masculinizations can be obtained in most species using androgen treatments. To gain insight into the molecular effects of androgen-induced masculinization we characterized, in the rainbow trout, the gonadal expression profiles of 103 candidate genes involved in sex differentiation and early gametogenesis. The androgen treatment (I I P-hydroxyandrostenedione, 10 mg/kg of food for 3 months) was administered in a genetic all-female population. Gonads were sampled at different time points in genetic all-male and all-female control populations and in the androgen-treated group. Gene expression profiles were recorded by real-time RT-PCR and biological samples and gene expressions were compared using a global clustering analysis. This analysis revealed that masculinization with androgens acts firstly by repressing granulosa cell related genes, including genes involved in ovarian differentiation (foxl2a, fst, cyp19a1a), and subsequently by repressing genes important for early oogenesis (gdj9, bcl2lb, fancl, gcl, fshb, lhb, sox23, sox24, nup62 and vtgr). However, this masculinizing treatment did not induce a testicular differentiation similar to what was observed in the control male population. This was especially noticeable for many Leydig cell genes encoding proteins involved in steroidogenesis or its control (hsd3b1, star, cyp17a1, cyp11b2.1 and nr5a1b) that were down-regulated in the androgen-treated group. Concomitantly some Sertoli cells marker genes were up-regulated by the androgen treatment (sox9a.1, nr0b1, cldn11, dmrt1) whereas others were down-regulated (amh, sox9a.2), suggesting a partial differentiation of the Sertoli cell lineage. Overall, this suggests that the crucial step of this masculinization process is the de-differentiation of the granulosa cells. (C) 2008 Elsevier Inc. All rights reserved.
Published: 2008

27. No early gender effects on energetic status and life history in a salmonid

Author: Thomas Régnier, Ayaka Yano, Joëlle Chat, Valérie Bolliet, Yann Guiguen, Jacques Labonne, Ecologie Comportementale et Biologie des Populations de Poissons (ECOBIOP), Institut National de la Recherche Agronomique (INRA)-Université de Pau et des Pays de l'Adour (UPPA), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), All authors were funded by the French National Institute of Agronomic Research (INRA) and the Atlantic Aquatic Resource Conservation (AARC) INTERREG project, and Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA)
Subjects: Long lasting, [SDV]Life Sciences [q-bio], gène déterminant majeur du sexe, salmo trutta, truite de mer, émergence, sexe, reproduction, Brown trout, mâle, gender effect, brown trout, poisson, femelle, genre, Salmo, Life history, lcsh:Science, Organism, ComputingMilieux_MISCELLANEOUS, gender effects, salmonidae, Multidisciplinary, biology, Ecology, Biology (Whole Organism), biology.organism_classification, trait de vie, dimorphisme sexuel, Sexual dimorphism, Sexual selection, Metabolic rate, lcsh:Q, sdy, energetic status, emergence from gravel, différenciation sexuelle, analyse génétique, Research Article, Demography
Abstract: Throughout an organism's early development, variations in physiology and behaviours may have long lasting consequences on individual life histories. While a large part of variation in critical life-history transitions remains unexplained, a significant proportion may be caused by early gender effects as part of gender-specific life histories shaped by sexual selection. In this study, we investigated the presence of early gender effects on the timing of emergence from gravel and the energetic status of brown trout ( Salmo trutta ) early stages. To investigate this question, individual measures of emergence timing, metabolic rate and energetic content were coupled for the first time with the use of a recent genetic marker for sdY (sexually dimorphic on the Y-chromosome), a master sex-determining gene. Our results show that gender does not influence the energetic content of emerging juveniles or their emergence timing. These findings suggest that gender differences may appear later throughout salmonid life history and that selective pressures associated with the critical period of emergence from gravel may shape early life-history traits similarly in both males and females.
Published: 2015

28. Localization of steroidogenic enzymes and Foxl2a in the gonads of mature zebrafish [i](Danio rerio)[/i]

Author: Jean-Marc Porcher, François Brion, Nathalie Hinfray, Morgane Caulier, Edith Chadili, Benjamin Piccini, Cyril Turies, Yann Guiguen, Direction des Risques Chroniques, Pole VIVA, Unité d'écotoxicologie in vitro et in vivo, Institut National de l'Environnement Industriel et des Risques, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), 190 AP 2010-2013, French National Ministry of Ecology and Sustainable Development, and Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA)
Subjects: Forkhead Box Protein L2, Male, Physiology, Somatic cell, Steroidogenic enzymes, 17-Alpha-hydroxylase, [SDV]Life Sciences [q-bio], Danio, Ovary, Foxl2a, Biochemistry, Isozyme, 03 medical and health sciences, Aromatases, 0302 clinical medicine, Aromatase, Cytochrome P-450 Enzyme System, Testis, medicine, Animals, Gonads, Molecular Biology, Gene, Zebrafish, 030304 developmental biology, 0303 health sciences, biology, Steroid 17-alpha-Hydroxylase, Forkhead Transcription Factors, Zebrafish Proteins, biology.organism_classification, Molecular biology, Immunohistochemistry, Cell biology, Isoenzymes, 20 lyase, medicine.anatomical_structure, Microscopy, Fluorescence, CYP17A1, biology.protein, Steroid 11-beta-Hydroxylase, Female, 11-Beta-hydroxylase, 030217 neurology & neurosurgery
Abstract: In zebrafish, the identification of the cells expressing steroidogenic enzymes and their regulators is far from completely fulfilled though it could provide crucial information on the elucidation of the role of these enzymes. The aim of this study was to better characterize the expression pattern of steroidogenic enzymes involved in estrogen and androgen production (Cyp17-I, Cyp11c1, Cyp19a1a and Cyp19a1b) and one of their regulators (Foxl2a) in zebrafish gonads. By using immunohistochemistry, we localized the steroid-producing cells in mature zebrafish gonads and determined different expression patterns between males and females. All these steroidogenic enzymes and Foxl2a were detected both in the testis and ovary. In the testis, they were all localized both in Leydig and germ cells except Cyp19a1b which was only detected in germ cells. In the ovary, Cyp17-I, Cyp19a1a and Foxl2a were immunolocalized in both somatic and germ cells while Cyp19a1b was only detected in germ cells and Cyp11c1 in somatic cells. Moreover, Cyp19a1a and Foxl2a did not display exactly the same patterns of spatial localization but their expressions were correlated suggesting a possible regulation of cyp19a1a gene by Foxl2a in zebrafish. Comparative analysis revealed a dimorphic expression of Cyp11c1, Cyp19a1a, Cyp19a1b and Foxl2a between males and females. Overall, our study provides a detailed description of the expression of proteins involved in the biosynthesis of steroidal hormones at the cellular scale within gonads, which is critical to further elucidating the intimate roles of the enzymes and the use of the zebrafish as a model in the field of endocrinology.
Published: 2015

29. Meta-analysis of microarray data of rainbow trout fry gonad differentiation modulated by ethynylestradiol

Author: Patrick Kestemont, Yann Guiguen, Bertrand De Meulder, Florence Le Gac, Sophie Depiereux, Eric Depiereux, Michael Pierre, Raphaël Helaers, Research Unit in Environmental and Evolutionary Biology (URBE-NARILIS), Université de Namur [Namur], Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Research Unit in Molecular Biology (URBM-NARILIS), FRS-FNRS Fonds de la Recherche Scientifique (Belgique), Université de Namur [Namur] (UNamur), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), and ProdInra, Archive Ouverte
Subjects: TBX1, Sex Differentiation, Microarray, [SDV]Life Sciences [q-bio], lcsh:Medicine, Biology, Ethinyl Estradiol, gonade, influence de l'exposition, reproduction, mâle, poisson, femelle, Gene expression, Animals, lcsh:Science, Gonads, éthynyloestradiol, Oligonucleotide Array Sequence Analysis, salmonidae, Genetics, Multidisciplinary, Sexual differentiation, Microarray analysis techniques, inversion sexuelle, lcsh:R, Sex reversal, cycle cellulaire, [SDV] Life Sciences [q-bio], analyse microarray, Oncorhynchus mykiss, lcsh:Q, Rainbow trout, Toxicogenomics, facteur de transcription, différenciation sexuelle, truite arc en ciel, expression des gènes, Research Article
Abstract: Sex differentiation in fish is a highly labile process easily reversed by the use of exogenous hormonal treatment and has led to environmental concerns since low doses of estrogenic molecules can adversely impact fish reproduction. The goal of this study was to identify pathways altered by treatment with ethynylestradiol (EE2) in developing fish and to find new target genes to be tested further for their possible role in male-to-female sex transdifferentiation. To this end, we have successfully adapted a previously developed bioinformatics workflow to a meta-analysis of two datasets studying sex reversal following exposure to EE2 in juvenile rainbow trout. The meta-analysis consisted of retrieving the intersection of the top gene lists generated for both datasets, performed at different levels of stringency. The intersecting gene lists, enriched in true positive differentially expressed genes (DEGs), were subjected to over-representation analysis (ORA) which allowed identifying several statistically significant enriched pathways altered by EE2 treatment and several new candidate pathways, such as progesterone-mediated oocyte maturation and PPAR signalling. Moreover, several relevant key genes potentially implicated in the early transdifferentiation process were selected. Altogether, the results show that EE2 has a great effect on gene expression in juvenile rainbow trout. The feminization process seems to result from the altered transcription of genes implicated in normal female gonad differentiation, resulting in expression similar to that observed in normal females (i.e. the repression of key testicular markers cyp17a1, cyp11b, tbx1), as well as from other genes (including transcription factors) that respond specifically to the EE2 treatment. The results also showed that the bioinformatics workflow can be applied to different types of microarray platforms and could be generalized to (eco)toxicogenomics studies for environmental risk assessment purposes.
Published: 2015

30. RAD-seq mapping of spontaneous masculinization in XX doubled haploid rainbow trout lines

Author: René Guyomard, Yann Guiguen, Maria Bernard, Aurélie Charlet, Nicolas Dechamp, Caroline Hervet, Céline Chantry-Darmon, Francine Krieg, Edwige Quillet, Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Laboratoire d'Analyse Génétique pour les Espèces Animales (LABOGENA), Institut National de la Recherche Agronomique (INRA), ANR, and Institut National de la Recherche Agronomique (INRA)-AgroParisTech
Subjects: salmonidae, oncorhynchus mykiss, détermination du sexe, déterminisme génétique, masculinisation, inversion sexuelle, sex determinism, [SDV]Life Sciences [q-bio], linkage map, reproduction, mâle, poisson, femelle, salmonids, truite arc en ciel
Abstract: Sex in rainbow trout is primarily determined by a genetic male heterogametic system encoded by a master sex-determining gene sdY. However, the occurrence of spontaneous phenotypic males in all-female doubled haploid (DH) gynogenetic populations suggested that multiple sex-influencing loci could exist in trout as found in other species (tilapia, common carp, Atlantic salmon, zebrafish). In order to get further insight in the genetic determinism of this un-expected masculinization, we crossed a DH XX spontaneously sex-reversed male with a normal female. Two F1 females were then reproduced by mitogynogenesis and their DH progeny (n=168) were genotyped using RAD sequencing. A genome scan of 9500 markers was obtained resulting in a linkage map of 2860 cM. Several putative QTL regions have been identified suggesting that more than one mutation could be involved in the observed deviation from the strict XX/XY sex determinism system.
Published: 2014

31. High temperature increases the masculinization rate of the all-female (XX) rainbow trout 'mal' population

Author: Edwige Quillet, Karina Valdivia, Brigitte Mourot, Alexis Fostier, Jean-Nicolas Volff, Delphine Galiana-Arnoux, Louise Helary, Elodie Jouanno, Yann Guiguen, René Guyomard, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, Institut de Génomique Fonctionnelle de Lyon (IGFL), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA), Génétique Animale et Biologie Intégrative (GABI), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, French National Research Agency (‘‘Sexytrout’’ project, ANR-2008-GENM-031) - Fellowship from the 'Ministère de la Recherche et de l'Enseignement Supérieur', Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-École normale supérieure - Lyon (ENS Lyon), and École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL)
Subjects: Male, Heredity, Hot Temperature, Sex Differentiation, gonade, mâle, poisson, Morphogenesis, Biologie de la reproduction, Genetics, salmonidae, 0303 health sciences, education.field_of_study, Reproductive Biology, oncorhynchus mykiss, détermination du sexe, Multidisciplinary, Sexual Differentiation, 04 agricultural and veterinary sciences, Trout, Phenotypes, medicine.anatomical_structure, Phenotype, Medicine, Female, Sex ratio, différenciation sexuelle, Research Article, Gonad, masculinisation, glycogène, Science, Population, Zoology, Marine Biology, Biology, reproduction, 03 medical and health sciences, femelle, taux de mortalite, température, Sex-determination system, medicine, Animals, 14. Life underwater, Sex Ratio, education, 030304 developmental biology, Evolutionary Biology, Sexual differentiation, Population Biology, Models, Genetic, Hatching, Biology and Life Sciences, Fisheries Science, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex Determination, Sex Determination Processes, biology.organism_classification, Mutation, 040102 fisheries, 0401 agriculture, forestry, and fisheries, Rainbow trout, Population Genetics, Developmental Biology, truite arc en ciel
Abstract: Salmonids are generally considered to have a robust genetic sex determination system with a simple male heterogamety (XX/XY). However, spontaneous masculinization of XX females has been found in a rainbow trout population of gynogenetic doubled haploid individuals. The analysis of this masculinization phenotype transmission supported the hypothesis of the involvement of a recessive mutation (termed mal). As temperature effect on sex differentiation has been reported in some salmonid species, in this study we investigated in detail the potential implication of temperature on masculinization in this XX mal-carrying population. Seven families issued from XX mal-carrying parents were exposed from the time of hatching to different rearing water temperatures ((8, 12 and 18°C), and the resulting sex-ratios were confirmed by histological analysis of both gonads. Our results demonstrate that masculinization rates are strongly increased (up to nearly two fold) at the highest temperature treatment (18°C). Interestingly, we also found clear differences between temperatures on the masculinization of the left versus the right gonads with the right gonad consistently more often masculinized than the left one at lower temperatures (8 and 12°C). However, the masculinization rate is also strongly dependent on the genetic background of the XX mal-carrying families. Thus, masculinization in XX mal-carrying rainbow trout is potentially triggered by an interaction between the temperature treatment and a complex genetic background potentially involving some part of the genetic sex differentiation regulatory cascade along with some minor sex-influencing loci. These results indicate that despite its rather strict genetic sex determinism system, rainbow trout sex differentiation can be modulated by temperature, as described in many other fish species.
Published: 2014

32. 17β-Estradiol Treatment Decreases Steroidogenic Enzyme Messenger Ribonucleic Acid Levels in the Rainbow Trout Testis*

Author: Hocine Mecherouki, Marina Govoroun, Yann Guiguen, Oonagh Marie McMeel, and Terry J. Smith
Subjects: Male, endocrine system, medicine.medical_specialty, 3-Hydroxysteroid Dehydrogenases, medicine.drug_class, Gene Expression Regulation, Enzymologic, Aromatase, Endocrinology, Internal medicine, Testis, Gene expression, medicine, Animals, Cholesterol Side-Chain Cleavage Enzyme, RNA, Messenger, Messenger RNA, Sexual differentiation, Estradiol, biology, Cholesterol side-chain cleavage enzyme, Steroid 17-alpha-Hydroxylase, Lyase, Mechanism of action, Estrogen, Oncorhynchus mykiss, Steroid Hydroxylases, biology.protein, Steroid 11-beta-Hydroxylase, Steroids, medicine.symptom
Abstract: In fish, estrogens are well known for their involvement in ovarian differentiation and have been shown to be very potent feminizing agents when administrated in vivo during early development. However, the mechanism of action of exogenous estrogens is poorly understood. We report here on the feminizing effects of estrogen treatment on the testicular levels of some steroidogenic enzyme messenger RNAs [mRNAs; cholesterol side-chain cleavage (P450scc), 17-hydroxylase/lyase (P450c17), 3beta-hydroxysteroid dehydrogenase (3betaHSD), 11beta-hydroxylase (P45011beta), and aromatase (P450aro)] in the rainbow trout, Oncorhynchus mykiss. Treatment was carried out by dietary administration of 17beta-estradiol (E(2); dosage of 20 mg/kg diet) to a genetically all male population. Steroidogenesis in the differentiating testis was demonstrated to be strongly altered by E(2), as this treatment resulted in considerable decrease in P450c17, 3betaHSD, and P45011beta mRNAs after only 10 days of treatment. In contrast, P450scc and P450aro mRNA levels were unaffected by E(2), with P450scc mRNA levels remaining unaltered and P450aro not stimulated by this feminizing estrogen treatment. To better characterize this E(2) effect, the same treatment was applied on postdifferentiating males, and roughly the same expression pattern was detected with a considerable decrease in testicular P450c17, 3betaHSD, and P45011beta mRNAs and a significant, but reduced, decrease in P450scc mRNA. In the interrenal, these steroidogenic enzyme mRNAs were not significantly affected by this E(2) treatment, except for a slight, but significant, decrease in P450scc mRNA. These results clearly demonstrate that estrogens have profound effects on testicular steroidogenesis and that they are acting specifically on the testis by decreasing mRNA steady state levels of many steroidogenic enzyme genes. The decrease in P45011beta mRNA, and thus inhibition of the synthesis of testicular 11-oxygenated androgens, may be an important step required for the active feminization of these genetic males.
Published: 2001

33. Search for genes involved in the temperature-induced gonadal sex differentiation in the tilapia,Oreochromis niloticus

Author: Marina Govoroun, Jean-François Baroiller, Alexis Fostier, Yann Guiguen, Helena D'Cotta, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), and Institut National de la Recherche Agronomique (INRA)
Subjects: Male, medicine.medical_specialty, Candidate gene, DNA, Complementary, Sex Differentiation, medicine.drug_class, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Biology, 03 medical and health sciences, chemistry.chemical_compound, Internal medicine, medicine, Animals, L52 - Physiologie animale - Croissance et développement, [INFO]Computer Science [cs], Aromatase, Gonads, DNA Primers, 030304 developmental biology, Regulation of gene expression, 0303 health sciences, Differential display, Sexual differentiation, Base Sequence, Temperature, Gene Expression Regulation, Developmental, Cell Differentiation, 04 agricultural and veterinary sciences, General Medicine, Sex Determination Processes, biology.organism_classification, Androgen, Oreochromis, Phenotype, Endocrinology, chemistry, 040102 fisheries, biology.protein, Steroid 11-beta-Hydroxylase, 0401 agriculture, forestry, and fisheries, 11-Ketotestosterone, Female, Animal Science and Zoology, Tilapia
Abstract: International audience; In the tilapia, Oreochromis niloticus, sex is determined by genetic factors (XX/XY) but temperature can also influence the gonadal sex differentiation. Elevated temperatures of 35 degreesC can generate functional male phenotypes if applied before and during sexual differentiation. The genes and mechanisms by which temperature acts on the cascade leading to sex differentiation have been investigated. Two strategies have been followed: 1) Search for novel genes by differential display, and 2) Expression studies of candidate genes. Genetically all-female and all-male progenies were reared at 27 degreesC (natural temperature) and at 35 degreesC (masculinizing treatment) and gonads dissected. Using differential display, we isolated a 300 bp cDNA (MM20C) from temperature-masculinized females. Virtual northern analysis revealed a 1.2 kb transcript in 35 degreesC treated females and males, but hardly any expression in natural females (27 degreesC). Semi-quantitative RT-PCR established a several-fold increase in MM20C expression in 35 degreesC masculinized fry. Elevated expression was observed in natural males (27 degreesC) with higher levels detected in those reared at 35 degreesC. Furthermore, we have analyzed as a candidate gene the P450 11 beta -hydroxylase, an important androgen steroidogenic enzyme. Low levels of expression were found in natural males. This coincides with low concentrations of 11 ketotestosterone in the gonads before and during gonadal sex differentiation. Higher expression levels of 11 beta -hydroxylase were detected in male gonads at 35 degreesC but levels in phenotypic males were similar to those found for natural females. Previous results reported that expression of aromatase is repressed by masculinizing treatments. Our study demonstrated that masculinizing-temperature can also stimulate the expression of other gene(s).
Published: 2001

34. Expression of cytochrome P45011β (11β-hydroxylase) gene during gonadal sex differentiation and spermatogenesis in rainbow trout, Oncorhynchus mykiss

Author: Jean-Jacques Lareyre, Shaojun Liu, Marina Govoroun, Marie-José Ricordel, Oonagh Marie McMeel, Helena D'Cotta, Yoshitaka Nagahama, Terry J. Smith, and Yann Guiguen
Subjects: Male, Sex Differentiation, Endocrinology, Diabetes and Metabolism, Clinical Biochemistry, poisson (animal), Cytochrome P450, Testicle, Biochemistry, Stéroïde, Endocrinology, Testis, Gene expression, Japanese eel, Cloning, Molecular, L53 - Physiologie animale - Reproduction, Spermatogénèse, Gene Expression Regulation, Developmental, Reverse transcription polymerase chain reaction, medicine.anatomical_structure, Oncorhynchus mykiss, Molecular Medicine, Female, endocrine system, medicine.medical_specialty, DNA, Complementary, animal structures, Molecular Sequence Data, Biology, Gene Expression Regulation, Enzymologic, Internal medicine, Complementary DNA, medicine, Animals, Amino Acid Sequence, Northern blot, Spermatogenesis, Molecular Biology, DNA Primers, Sexual differentiation, Base Sequence, urogenital system, Cell Biology, biology.organism_classification, Molecular biology, Steroid 11-beta-Hydroxylase, Rainbow trout
Abstract: Androgens and especially 11-oxygenated androgens are known to be potent masculinizing steroids in fish. As a first step to study their physiological implication in gonadal sex differentiation in fish, we cloned a testicular cytochrome P450(11beta) (11beta-hydroxylase) cDNA in the rainbow trout, Oncorhynchus mykiss. We isolated a 1882 bp P450(11beta) cDNA (rt11betaH2, AF217273) which contains an open reading frame encoding a 552 putative amino acids protein. This sequence was highly homologous (98% in nucleotides and 96.5% in amino acids) to another rainbow trout P450(11beta) sequence (AF179894) and also to a Japanese eel P450(11beta) (68% in amino acids). Northern blot analysis detected a single transcript of 2 kb which was highly expressed in the testis (stage II) and to a lesser degree in the anterior kidney (containing the interrenal tissue). No signal was detected in the posterior kidney, brain, liver, skin, intestine and heart. In the testis this transcript was highly expressed at the beginning of spermatogenesis (stages I and II), followed by a decrease during late spermatogenesis (stages III to V). By semi-quantitative reverse transcription polymerase chain reaction, P450(11beta) expression during gonadal differentiation was estimated to be at least 100 times higher in male than in female gonads. This difference was first detected at 55 days post-fertilization (dpf), i.e. 3 weeks before the first sign of histological sex differentiation, and was sustained long after differentiation (127 dpf). Specific P450(11beta) gene expression was also demonstrated before testis differentiation (around 50 dpf) using virtual Northern blot, with no expression detected in female differentiating gonads. From these results, and also based on the already known actions of 11-oxygenated androgens in testicular differentiation in fish, it is now suggested that P450(11beta) gene expression is a key factor for the testicular differentiation in rainbow trout.
Published: 2000

35. Sex differentiation in an all-female (XX) rainbow trout population with a genetically governed masculinization phenotype

Author: Elodie Jouanno, Béatrice Collin, Yann Guiguen, Delphine Galiana-Arnoux, Brigitte Mourot, Louise Helary, Jean-Nicolas Volff, Alexis Fostier, René Guyomard, Chantal Cauty, Karina Valdivia, Pablo Rault, Edwige Quillet, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA)-IFR140, Institut de Génomique Fonctionnelle de Lyon (IGFL), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Génétique Animale et Biologie Intégrative (GABI), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, ANR-2008-GENM-031, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-École normale supérieure - Lyon (ENS Lyon), and École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL)
Subjects: Male, Embryology, medicine.medical_specialty, Aging, endocrine system, X Chromosome, Endocrinology, Diabetes and Metabolism, Population, salmonid, sex determination, Biology, gametogenesis, Andrology, histology, 03 medical and health sciences, 0302 clinical medicine, Internal medicine, medicine, Animals, education, Gonads, Gametogenesis, 030304 developmental biology, 0303 health sciences, education.field_of_study, Sex Characteristics, Sexual differentiation, [SDV.BA]Life Sciences [q-bio]/Animal biology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, sex inversion, Phenotype, Endocrinology, medicine.anatomical_structure, Germ Cells, Oncorhynchus mykiss, sex differentiation, Rainbow trout, Female, mutation, 030217 neurology & neurosurgery, Heterogametic sex, Germ cell, Biomarkers, Developmental Biology, Sex characteristics
Abstract: Sex determination is known to be male heterogametic in the rainbow trout, Oncorhynchus mykiss; however, scattered observations that deviate from this rather strict genetic control have been reported. Here, we provide a detailed morphological and histological characterization of the gonadal differentiation and development (from 43 days postfertilization to 11 months of age) in an all-female (XX) population with a genetically governed masculinization phenotype. In comparison with control males and females, the gonadal differentiation in these animals was characterized by many perturbations, including significantly fewer germ cells. This decrease in germ cells was confirmed by the significantly decreased expression of 2 germ cell maker genes (vasa and sycp3) in the masculinized XX populations as compared with the control females and control males. Although only a proportion of the total adult population was partially or fully masculinized, this early differentiating phenotype affected nearly all the sampled animals. This suggests that the adult masculinization phenotype is the consequence of an early functional imbalance in ovarian differentiation in the entire population. We hypothesize that the lower number of germ cells that we observed in this population could be one cause of their masculinization.
Published: 2013

36. Gonadal in Vitro Androstenedione Metabolism and Changes in Some Plasma and Gonadal Steroid Hormones during Sex Inversion of the Protandrous Sea Bass, Lates calcarifer

Author: A. Benett, Bernard Jalabert, A. Fostier, and Yann Guiguen
Subjects: Male, endocrine system, medicine.medical_specialty, 3-Hydroxysteroid Dehydrogenases, Gonad, 17-Hydroxysteroid Dehydrogenases, Metabolite, Radioimmunoassay, Ovary, Estrone, chemistry.chemical_compound, Endocrinology, Internal medicine, Testis, medicine, Animals, Hermaphroditic Organisms, Testosterone, Androstenedione, Aromatase, Sea bass, Chromatography, High Pressure Liquid, Estradiol, biology, Hydroxysteroid Dehydrogenases, Sex Determination Processes, medicine.anatomical_structure, chemistry, biology.protein, 11-beta-Hydroxysteroid Dehydrogenases, Bass, Female, Animal Science and Zoology, Seasons
Abstract: Steroidogenesis in the gonad of the protandrous sea bass, Lates calcarifer, was examined in vitro in spermiating testis, previtellogenic ovary, and transitional gonads. Gonadal tissues were incubated with tritiated androstenedione. Metabolites were analyzed by thin-layer chromatography, high-performance liquid chromatography, microchemical reactions, and crystallization to constant specific activity. 17 beta-Hydroxysteroid dehydrogenase, 5 beta-reductase, and 3 alpha-hydroxysteroid dehydrogenase activities were found in all of the sex types. On the other hand, 11 beta-hydroxysteroid dehydrogenase and 11 beta-hydroxylase activities were found only when testicular tissue was present, i.e., in testis and early transitional gonad. A low aromatase activity leading to estrone synthesis was detected in the previtellogenic ovary. In late transitional gonads, a major metabolite (metabolite X) was suggestively identified as a 3-ester of 17 beta-estradiol according to its chemical and immunological characteristics. Levels of 17 beta-estradiol (E2), the metabolite X, testosterone (T), and 11-ketotestosterone (11KT) were also measured by radioimmunoassay in plasma, before (January and February) and during (March and April) the sex inversion process. Plasma E2 was virtually undetectable (means below 25 pg/ml), although higher levels of metabolite X were found in transitional fish (485 +/- 432 pg/ml in March). Throughout this period, plasma levels of T and 11KT and the androgens/estrogens ratio were significantly higher in males than in transitional fish, where these levels decreased during the sex inversion period. The level of in vitro synthesis of metabolite X was high in transitional gonads, but their concentrations were very low (0.07 +/- 0.09 ng of equivalent E2/g in transitional gonads against 0.22 +/- 0.37 ng of equivalent E2/g in testes and 2.16 +/- 2.7 ng of equivalent E2/g in ovaries).
Published: 1995

37. New insights regarding gonad development in European eel: evidence for a direct ovarian differentiation

Author: Benjamin Geffroy, Alexis Fostier, Agnès Bardonnet, Yann Guiguen, Ecologie Comportementale et Biologie des Populations de Poissons (ECOBIOP), Institut National de la Recherche Agronomique (INRA)-Université de Pau et des Pays de l'Adour (UPPA), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), The project was supported by funding from the Office National de l’Eau et des Milieux Aquatiques (ONEMA) and by the Interreg IV B Atlantic area transnational program (European Regional Development Fund), in the context of the Arc Atlantic Resource Conservation program (AARC), and Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )
Subjects: 0106 biological sciences, Male, Sex Differentiation, Physiology, european eel, Sex assessment, 01 natural sciences, Biochemistry, gonad development, Aromatase, ComputingMilieux_MISCELLANEOUS, 0303 health sciences, education.field_of_study, Sex Characteristics, Eels, biology, Reverse Transcriptase Polymerase Chain Reaction, [SDV.BA]Life Sciences [q-bio]/Animal biology, General Medicine, medicine.anatomical_structure, Female, France, Development of the gonads, Estuaries, medicine.medical_specialty, endocrine system, Gonad, growth, Population, Ovary, Aquatic Science, Real-Time Polymerase Chain Reaction, 010603 evolutionary biology, Statistics, Nonparametric, Andrology, 03 medical and health sciences, Internal medicine, medicine, sex, Animals, 14. Life underwater, education, Gonads, 030304 developmental biology, DNA Primers, Sexual differentiation, Histology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex Determination Processes, Endocrinology, biology.protein
Abstract: International audience; In European eel, it has been proposed that the undifferentiated gonad would develop into either an intersexual stage (Syrski organ) or directly into an ovary. The Syrski organ could then develop into either an ovary or a testis. In the present study, glass eels were raised until they reached a minimum size of 29 cm for histological sex assessment. In addition, some undifferentiated individuals with size encompassing 15–28 cm were sampled in a female-biased population (Oir River). We also investigated aromatase gene expression, which is known to be involved in the process of fish sex differentiation. Gonad histology revealed that intersexual eels were characterized by a small number of oocytes within a predominant testis-like structure. Males were significantly smaller than intersexual eels, which suggests that all males do not necessarily pass through an intermediate intersexual stage. Aromatase transcript levels in intersexual eels gonads and testes were similar but significantly lower than in ovaries and were comparable between ovaries and undifferentiated gonads from the females-biased population. In addition, condition factor was lower in female than in intersexual individuals. Together, these results provide evidence that ovaries would not develop from the Syrski organ
Published: 2012

38. An immune-related gene evolved into the master sex-determining gene in Rainbow-trout, Oncorhynchus mykiss

Author: Edwige Quillet, Christophe Klopp, Ayaka Yano, Alexis Fostier, Cédric Cabau, Elodie Jouanno, Barbara Nicol, Olivier Bouchez, Yann Guiguen, René Guyomard, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA)-IFR140, Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Unité de Biométrie et Intelligence Artificielle (UBIA), Institut National de la Recherche Agronomique (INRA), Laboratoire de Génétique Cellulaire (LGC), Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA), ANR (SVSE 7 2011, project SDS), Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées
Subjects: Male, 0106 biological sciences, Molecular Sequence Data, Locus (genetics), Biology, 010603 evolutionary biology, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Sequence Homology, Nucleic Acid, Y Chromosome, biology.animal, Testis, Animals, Amino Acid Sequence, Gene, 030304 developmental biology, Genetics, 0303 health sciences, Sexual differentiation, Base Sequence, Agricultural and Biological Sciences(all), Biochemistry, Genetics and Molecular Biology(all), Vertebrate, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex Determination Processes, Transgenesis, Sexual dimorphism, Testis determining factor, Oncorhynchus mykiss, Interferon Regulatory Factors, Female, General Agricultural and Biological Sciences, Interferon regulatory factors
Abstract: Chantier qualité GA; Since the discovery of Sry in mammals [ 1 and 2], few other master sex-determining genes have been identified in vertebrates [ 3, 4, 5, 6 and 7]. To date, all of these genes have been characterized as well-known factors in the sex differentiation pathway, suggesting that the same subset of genes have been repeatedly and independently selected throughout evolution as master sex determinants [ 8 and 9]. Here, we characterized in rainbow trout an unknown gene expressed only in the testis, with a predominant expression during testicular differentiation. This gene is a male-specific genomic sequence that is colocalized along with the sex-determining locus. This gene, named sdY for sexually dimorphic on the Y chromosome, encodes a protein that displays similarity to the C-terminal domain of interferon regulatory factor 9. The targeted inactivation of sdY in males using zinc-finger nuclease induces ovarian differentiation, and the overexpression of sdY in females using additive transgenesis induces testicular differentiation. Together, these results demonstrate that sdY is a novel vertebrate master sex-determining gene not related to any known sex-differentiating gene. These findings highlight an unexpected evolutionary plasticity in vertebrate sex determination through the demonstration that master sex determinants can arise from the de novo evolution of genes that have not been previously implicated in sex differentiation.
Published: 2012

39. Expression profiling of Wnt signaling genes during gonadal differentiation and gametogenesis in rainbow trout

Author: Barbara Nicol, Yann Guiguen, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), SCRIBE Campus de Beaulieu, Institut National de la Recherche Agronomique (INRA), Institut Fédératif de Recherche - Génétique Fonctionnelle Agronomie et Santé (IFR 140 GFAS), Plateforme Génomique Santé Biogenouest®, Brittany province (Region Bretagne), INRA Institute (PHASE department), European Community [222719-LIFECYCLE], Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Brittany province (Région Bretagne) - INRA Institute (PHASE department), and European Project: 222719,EC:FP7:KBBE,FP7-KBBE-2007-2A,LIFECYCLE(2009)
Subjects: Male, Embryology, medicine.medical_specialty, endocrine system, gamétogenèse, Sex Differentiation, Endocrinology, Diabetes and Metabolism, [SDV]Life Sciences [q-bio], Ovary, Biology, Real-Time Polymerase Chain Reaction, Oogenesis, Gametogenesis, 03 medical and health sciences, 0302 clinical medicine, Internal medicine, Testis, medicine, Animals, RSPO1, Wnt Signaling Pathway, In Situ Hybridization, 030304 developmental biology, teleost fish, 0303 health sciences, Sexual differentiation, urogenital system, Wnt signaling pathway, rainbow trout, Wnt signaling, GENOMIQUE, Cell biology, Gene expression profiling, Endocrinology, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Oncorhynchus mykiss, Female, Development of the gonads, Developmental Biology, SALMONIDE
Abstract: Wnt signaling plays major roles in various processes, including ovarian differentiation and development in mammals. In order to explore its potential implication during gonadal development in a nonmammalian vertebrate species, expression of Wnt signaling genes was investigated in rainbow trout during gonadal differentiation and gametogenesis. Multiple Wnt pathway genes were expressed and exhibited distinct expression patterns. In ovary, tcf7 was highly expressed during early differentiation, whereas no sexually dimorphic expression of rspo1 was detected. During later ovarian development, wnt11 was highly expressed in granulosa cells and oocytes suggesting an implication in folliculogenesis and oogenesis, whereas wnt9b was principally detected in granulosa cells. In testis, Wnt pathway genes were mostly expressed during early spermatogenesis. Overall, these present results suggest that Wnt signaling is implicated in multiple processes of male and female gonadal development and provide basis for future studies on Wnt signaling functions in teleost fish gonads.
Published: 2011

40. Characterization of rainbow trout gonad, brain and gill deep cDNA repertoires using a Roche 454-Titanium sequencing approach

Author: Cédric Cabau, Jérôme Lluch, Patrick Prunet, Barbara Nicol, Arianna Servili, François Moreews, Yann Guiguen, Christophe Klopp, Isabelle Le Guen, Olivier Kah, Jean-Jacques Lareyre, Olivier Bouchez, Aurélie Le Cam, Jérôme Montfort, Julien Bobe, Pierre-Yves Rescan, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), Institut Fédératif de Recherche - Génétique Fonctionnelle Agronomie et Santé (IFR 140 GFAS), Plateforme Génomique Santé Biogenouest®, Laboratoire de Génétique Cellulaire (LGC), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Unité de Biométrie et Intelligence Artificielle (UBIA), CNRS UMR6026 Interactions cellulaires et moléculaires (ICM), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Génopole, Plateforme Génomique Santé Biogenouest®. FRA., Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA), UMR 6026 Interactions cellulaires et moléculaires (ICM), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), AIP Bioressources INRA 2009 - Région Bretagne, Génopole Toulouse Midi-Pyrénées [Auzeville] (GENOTOUL), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Recherches Avicoles (URA), Interactions cellulaires et moléculaires (ICM), Unité de Biométrie et Intelligence Artificielle de Toulouse [Castanet-Tolosan] (UBIA), Institut National de la Recherche Agronomique (INRA)-Plateforme bioinformatique du GIS GENOTOUL - Génopole Toulouse Midi-Pyrénées, Système d'Information des GENomes des Animaux d'Elevage (SIGENAE), Scalable, Optimized and Parallel Algorithms for Genomics (GenScale), Inria Rennes – Bretagne Atlantique, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria)-GESTION DES DONNÉES ET DE LA CONNAISSANCE (IRISA-D7), Institut de Recherche en Informatique et Systèmes Aléatoires (IRISA), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Université de Bretagne Sud (UBS)-École normale supérieure - Rennes (ENS Rennes)-Institut National de Recherche en Informatique et en Automatique (Inria)-Télécom Bretagne-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Université de Bretagne Sud (UBS)-École normale supérieure - Rennes (ENS Rennes)-Institut National de Recherche en Informatique et en Automatique (Inria)-Télécom Bretagne-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche en Informatique et Systèmes Aléatoires (IRISA), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Université de Bretagne Sud (UBS)-École normale supérieure - Rennes (ENS Rennes)-Télécom Bretagne-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et Développement de Rennes (IGDR), Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Department of Biology [Univ. of Cadiz], University of Cadiz, Guiguen, Yann, GESTION DES DONNÉES ET DE LA CONNAISSANCE (IRISA-D7), Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Université de Bretagne Sud (UBS)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Rennes (ENS Rennes)-Institut National de Recherche en Informatique et en Automatique (Inria)-Télécom Bretagne-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-CentraleSupélec-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Université de Rennes (UNIV-RENNES)-CentraleSupélec-Institut de Recherche en Informatique et Systèmes Aléatoires (IRISA), Université de Rennes (UNIV-RENNES)-CentraleSupélec-Inria Rennes – Bretagne Atlantique, Institut National de Recherche en Informatique et en Automatique (Inria), Recherches Avicoles (SRA), Centre National de la Recherche Scientifique (CNRS)-IFR140-Université de Rennes 1 (UR1), Plateforme bioinformatique du GIS GENOTOUL - Génopole Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Université de Bretagne Sud (UBS)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Rennes (ENS Rennes)-Télécom Bretagne-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-CentraleSupélec, Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-IFR140-Centre National de la Recherche Scientifique (CNRS), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), European Project: 222719,EC:FP7:KBBE,FP7-KBBE-2007-2A,LIFECYCLE(2009), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT), Unité de Biométrie et Intelligence Artificielle (ancêtre de MIAT) (UBIA), and Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Gills, Male, muscle, animal diseases, [SDV]Life Sciences [q-bio], 0302 clinical medicine, poisson, Biologie de la reproduction, banque de données, [MATH]Mathematics [math], ComputingMilieux_MISCELLANEOUS, Genetics, Animal biology, salmonidae, 0303 health sciences, Differential display, Reproductive Biology, oncorhynchus mykiss, Contig, [SDV.BA]Life Sciences [q-bio]/Animal biology, Brain, High-Throughput Nucleotide Sequencing, General Medicine, Organ Specificity, cerveau, testicule, Female, expression des gènes, endocrine system, animal structures, salmonid, Computational biology, Biology, digestive system, DNA sequencing, 03 medical and health sciences, Complementary DNA, Biologie animale, séquençage, genomics, Animals, [INFO]Computer Science [cs], 14. Life underwater, Gonads, Gene, 030304 developmental biology, Whole genome sequencing, fish, cDNA library, urogenital system, Gene Expression Profiling, génome, ovaire, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sequence Analysis, DNA, branchie, rainbow-trout, transcriptome, next-generation sequencing, Rainbow trout, 030217 neurology & neurosurgery, truite arc en ciel
Abstract: International audience; Rainbow trout, Oncorhynchus mykiss, is an important aquaculture species worldwide and, in addition to being of commercial interest, it is also a research model organism of considerable scientific importance. Because of the lack of a whole genome sequence in that species, transcriptomic analyses of this species have often been hindered. Using next-generation sequencing (NGS) technologies, we sought to fill these informational gaps. Here, using Roche 454-Titanium technology, we provide new tissue-specific cDNA repertoires from several rainbow trout tissues. Non-normalized cDNA libraries were constructed from testis, ovary, brain and gill rainbow trout tissue samples, and these different libraries were sequenced in 10 separate half-runs of 454-Titanium. Overall, we produced a total of 3 million quality sequences with an average size of 328 bp, representing more than 1 Gb of expressed sequence information. These sequences have been combined with all publicly available rainbow trout sequences, resulting in a total of 242,187 clusters of putative transcript groups and 22,373 singletons. To identify the predominantly expressed genes in different tissues of interest, we developed a Digital Differential Display (DDD) approach. This approach allowed us to characterize the genes that are predominantly expressed within each tissue of interest. Of these genes, some were already known to be tissue-specific, thereby validating our approach. Many others, however, were novel candidates, demonstrating the usefulness of our strategy and of such tissue-specific resources. This new sequence information, acquired using NGS 454-Titanium technology, deeply enriched our current knowledge of the expressed genes in rainbow trout through the identification of an increased number of tissue-specific sequences. This identification allowed a precise cDNA tissue repertoire to be characterized in several important rainbow trout tissues. The rainbow trout contig browser can be accessed at the following publicly available web site (http://www.sigenae.org/).
Published: 2011

41. Sexual dimorphism in the brain aromatase expression and activity, and in the central expression of other steroidogenic enzymes during the period of sex differentiation in monosex rainbow trout populations

Author: Isabelle Anglade, Alexis Fostier, Yann Guiguen, Denise Vizziano-Cantonnet, Elisabeth Pellegrini, Marie-Madeleine Gueguen, Olivier Kah, Interactions cellulaires et moléculaires (ICM), Centre National de la Recherche Scientifique (CNRS)-IFR140-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA)-IFR140, IFR 140, CNRS, INRA, the Université of Rennes 1 (France), the European Community's Seventh Framework Programme (FP7/2007-2013) under grant agreement No. 30 222719 LIFECYCLE and the Comisión Sectorial de Investigación Científica (CSIC, Universidad de la República Oriental del Uruguay) of Uruguay., Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA), Laboratorio de Fisiologia de la Reproduccion y Ecologia de Peces, Instituto de Biologia, Facultad de Ciencias, Universidad de la Republica Oriental del Uruguay, Partenaires INRAE, and Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, Sex Differentiation, Steroidogenic enzymes, MESH: PROGENITOR CELLS, Estrogen receptor, Estrogen receptors, Endocrinology, Aromatase, 0303 health sciences, Sex Characteristics, Brain, 04 agricultural and veterinary sciences, medicine.anatomical_structure, Cholesterol, Receptors, Estrogen, MESH: ESTROGEN-RECEPTORS, Oncorhynchus mykiss, Female, Sex characteristics, MESH: ONCORHYNCHUS-MYKISS, Fish Proteins, MESH: GENE-TRANSCRIPTION, endocrine system, medicine.medical_specialty, Gonad, MESH: FISH ODONTESTHES-BONARIENSIS, MESH: ACUTE REGULATORY PROTEIN, Biology, MESH: DEVELOPMENTAL EXPRESSION, 03 medical and health sciences, MESH: MESSENGER-RNA EXPRESSION, Internal medicine, medicine, Animals, RNA, Messenger, 030304 developmental biology, Sexual differentiation, Cholesterol side-chain cleavage enzyme, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Sexual dimorphism, Fish, MESH: CYTOCHROME-P450 AROMATASE, MESH: DEVELOPING RAT-BRAIN, 040102 fisheries, biology.protein, 0401 agriculture, forestry, and fisheries, Animal Science and Zoology, Estrogen receptor alpha
Abstract: International audience; Using genetic monosex male and female rainbow trout populations, the potential sex differences in the central expression of estrogen receptors (esr1, esr2a, esr2b), brain aromatase (cyp19a1b) and some other steroidogenic enzymes was studied over the period of sex differentiation (from 35 to 63 dpf: days post-fertilization) using quantitative polymerase chain reaction (q-PCR). In addition, aromatase activity was evaluated during this period. The results indicated that brain aromatase (cyp19a1b) expression and activity showed a clear and significant sexually dimorphic pattern with higher levels in male brain between 35 and 53 dpf before the time of gonad morphological differentiation. At that time the expression of a key enzyme involved in the conversion of cholesterol into steroids, the cyp11a1 (p450scc), as well as the estrogen receptors were also sexually dimorphic. The dimorphism was lost from 56 dpf onwards. Transcription factors such as nr5a1b (sf1) and nr0b1 (dax1), but not foxl2a were also higher in males than in females. These results demonstrate that, before or during the early period of morphological gonad differentiation, the brain exhibits a clear sexual dimorphism with respect to the expression and activity of aromatase as well as of certain enzymes and factors involved in steroid synthesis as p450scc and sf1. The results suggest a higher potentiality to produce estrogens by male brains during sex differentiation time.
Published: 2010

42. Ovarian aromatase and estrogens: a pivotal role for gonadal sex differentiation and sex change in fish

Author: Francesc Piferrer, Ching-Fong Chang, Yann Guiguen, Alexis Fostier, Laboratoire de Physiologie et Génomique des Poissons (LPGP), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut Fédératif de Recherche - Génétique Fonctionnelle Agronomie et Santé (IFR 140 GFAS), Institute of Marine Sciences / Institut de Ciències del Mar [Barcelona] (ICM), Consejo Superior de Investigaciones Científicas [Spain] (CSIC), Department of Aquaculture, National Taiwan Ocean University (NTOU), Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), and Consejo Superior de Investigaciones Científicas [Madrid] (CSIC)
Subjects: Male, medicine.medical_specialty, endocrine system, Sex Differentiation, medicine.drug_class, Sex differentiation, 03 medical and health sciences, Sex change, Endocrinology, Aromatase, 5. Gender equality, Hermaphrodite, Internal medicine, medicine, sex change, Animals, gonads, Hermaphroditic Organisms, 14. Life underwater, Sexual Maturation, Gonads, cyp19a1a, Promoter Regions, Genetic, Transcription factor, 030304 developmental biology, Ovum, 0303 health sciences, Sexual differentiation, biology, Ovary, Fishes, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Estrogens, Forkhead Transcription Factors, 04 agricultural and veterinary sciences, Sex reversal, Sex Determination Processes, Estrogen, DMRT1 Gene, 040102 fisheries, biology.protein, 0401 agriculture, forestry, and fisheries, Animal Science and Zoology, Female
Abstract: 15 pages, 6 figures, 1 table.-- Special issue Fish Reproduction, The present review focuses on the roles of estrogens and aromatase (Cyp19a1a), the enzyme needed for their synthesis, in fish gonadal sex differentiation. Based on the recent literature, we extend the already well accepted hypothesis of an implication of estrogens and Cyp19a1a in ovarian differentiation to a broader hypothesis that would place estrogens and Cyp19a1a in a pivotal position to control not only ovarian, but also testicular differentiation, in both gonochoristic and hermaphrodite fish species. This working hypothesis states that cyp19a1a up-regulation is needed not only for triggering but also for maintaining ovarian differentiation and that cyp19a1a down-regulation is the only necessary step for inducing a testicular differentiation pathway. When considering arguments for and against, most of the information available for fish supports this hypothesis since either suppression of cyp19a1a gene expression, inhibition of Cyp19a1a enzymatic activity, or blockage of estrogen receptivity are invariably associated with masculinization. This is also consistent with reports on normal gonadal differentiation, and steroid-modulated masculinization with either androgens, aromatase inhibitors or estrogen receptor antagonists, temperature-induced masculinization and protogynous sex change in hermaphrodite species. Concerning the regulation of fish cyp19a1a during gonadal differentiation, the transcription factor foxl2 has been characterized as an ovarian specific upstream regulator of a cyp19a1a promoter that would co-activate cyp19a1a expression, along with some additional partners such as nr5a1 (sf1) or cAMP. In contrast, upstream factors potentially down-regulating cyp19a1a during testicular differentiation are still hypothetical, such as the dmrt1 gene, but their definitive characterization as testicular repressors of cyp19a1a would strongly strengthen the hypothesis that early testicular differentiation would need active repression of cyp19a1a expression
Published: 2008

43. Aromatase (P450arom) and 11beta-hydroxylase (P45011beta) genes are differentially expressed during the sex change process of the protogynous rice field eel, Monopterus albus

Author: Ji-Fang Liu, Shao-Jun Liu, Yann Guiguen, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), and Hunan Normal University
Subjects: Male, Sex Differentiation, Physiology, synbranchidae, Monopterus albus, Biochemistry, Homology (biology), Testis, sex change, Cluster Analysis, Aromatase, Cloning, Molecular, Phylogeny, 0303 health sciences, biology, Reverse Transcriptase Polymerase Chain Reaction, [SDV.BA]Life Sciences [q-bio]/Animal biology, rice field eel, 04 agricultural and veterinary sciences, General Medicine, Smegmamorpha, Female, Monopterus, medicine.medical_specialty, endocrine system, Molecular Sequence Data, Aquatic Science, Gene Expression Regulation, Enzymologic, 03 medical and health sciences, Sex change, Internal medicine, medicine, Animals, Gene, 030304 developmental biology, DNA Primers, P450 11β-hydroxylase, Sexual differentiation, Ovotestis, Base Sequence, Ovary, Cytochrome P450, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sequence Analysis, DNA, biology.organism_classification, Endocrinology, P450 aromatase, 040102 fisheries, biology.protein, 0401 agriculture, forestry, and fisheries, Steroid 11-beta-Hydroxylase, sense organs
Abstract: International audience; Steroids are known to play a crucial role in gonadal sex differentiation in many non-mammalian vertebrates, but also in the gonadal sex change of hermaphroditic teleosts. We investigated the expression of two genes encoding key steroidogenic enzymes, i.e., cytochrome P450 aromatase (P450arom) and cytochrome P45011 beta-hydroxylase (P45011 beta), during the sex change of the protogynous rice field eel, Monopterus albus. Using RT-PCR with degenerate primers, we cloned rice field eel homologous fragments for both genes (rcP450arom and rcP45011 beta) as indicated by the high level of homology with P450arom and P45011 beta sequences from various vertebrates. Gonadal expression of rcP450arom and rcP45011 beta mRNA levels were then assessed during the sex change by semi-quantitative RT-PCR and a real-time RT-PCR. rcP450arom was predominantly expressed in ovary, much less in ovotestis, and barely in testis. Conversely, P45011 beta was markedly up-regulated at the onset of testicular development. These findings underline that regulation of steroidogenesis is an important process in the sex change of protogynous rice field eel, and they clearly indicate that the concomitant down-regulation of P450arom and up-regulation of P45011 beta are of pivotal importance to the sex change of this species.
Published: 2008

44. Estrogen treatment up-regulates female genes but does not suppress all early testicular markers during rainbow trout male-to-female gonadal transdifferentiation

Author: Daniel Baron, Yann Guiguen, Denise Vizziano-Cantonnet, Sophie Mahé, Chantal Cauty, Alexis Fostier, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), and Universidad de la República
Subjects: Male, medicine.medical_specialty, endocrine system, Sex Differentiation, medicine.drug_class, Feminization (biology), salmonid, 030209 endocrinology & metabolism, Biology, feminization, FSHB, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Internal medicine, medicine, Animals, Cluster Analysis, Gonads, Molecular Biology, 030304 developmental biology, 0303 health sciences, Sexual differentiation, Leydig cell, Gene Expression Profiling, [SDV.BA]Life Sciences [q-bio]/Animal biology, Transdifferentiation, Gene Expression Regulation, Developmental, Estrogens, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sertoli cell, Gene expression profiling, medicine.anatomical_structure, Estrogen, Multigene Family, Oncorhynchus mykiss, Cell Transdifferentiation, Female, Biomarkers
Abstract: In non-mammalian vertebrates, estrogens are key players in ovarian differentiation, but the mechanisms by which they act remain poorly understood. The present study on rainbow trout was designed to investigate whether estrogens trigger the female pathway by activating a group of early female genes (i.e. cyp19a1, foxl2a, foxl2b, fst, bmp4, and fshb) and by repressing early testicular markers (i.e. dmrt1, nr0b1, sox9a1 and sox9a2). Feminization was induced in genetically all-male populations using 17α-ethynylestradiol (EE2, 20 mg/kg of food during 2 months). The expression profiles of 100 candidate genes were obtained by real-time RT-PCR and 45 expression profiles displayed a significant differential expression between control populations (males and females) and EE2-treated populations. These expression profiles were grouped in five temporally correlated expression clusters. The estrogen treatment induced most of the early ovarian differentiation genes (foxl2a, foxl2b, fst, bmp4, and fshb) and in particular foxl2a, which was strongly and quickly up-regulated. Simultaneously, Leydig cell genes, involved in androgen synthesis, as well as some Sertoli cell markers (amh, sox9a2) were strongly repressed. However, in contrast to our initial hypothesis, some genes considered as essential for mammalian and fish testis differentiation were not suppressed during the early process of estrogen-induced feminization (dmrt1, nr0b1, sox9a1 and pax2a) and some were even strongly up-regulated (nr0b1, sox9a1and pax2a). In conclusion, estrogens trigger male-to-female transdifferentiation by up-regulating most ovarian specific genes and this up-regulation appears to be crucial for an effective feminization, but estrogens do not concomitantly down-regulate all the testicular differentiation markers.
Published: 2008

45. The fate of the duplicated androgen receptor in fishes: a late neofunctionalization event ?

Author: Frédéric Brunet, Yann Guiguen, Bernard Haendler, Virginie Vlaeminck-Guillem, Isabelle Ahrens-Fath, Bastien Boussau, Veronique Douard, Vincent Laudet, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), Institut de Génomique Fonctionnelle de Lyon (IGFL), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-École normale supérieure - Lyon (ENS Lyon), Université de Lyon, Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Bayer Pharma AG [Berlin], École normale supérieure - Lyon (ENS Lyon), BioSciences Lyon-Gerland (BLG), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), and École normale supérieure de Lyon (ENS de Lyon)
Subjects: Male, Biodiversité et Ecologie, DNA Mutational Analysis, reproduction animale, GENOMIQUE, DUPLICATION DU GENOME, Osteoglossiformes, 0302 clinical medicine, poisson, Gene Duplication, Gene duplication, Cloning, Molecular, Zebrafish, Phylogeny, Genetics, évolution biologique, 0303 health sciences, Likelihood Functions, détermination du sexe, Genome, Fishes, Androgen-Insensitivity Syndrome, androgène, Receptors, Androgen, Neofunctionalization, récepteur, Research Article, Fish Proteins, Lineage (genetic), Evolution, Biology, Evolution, Molecular, Biodiversity and Ecology, 03 medical and health sciences, Phylogenetics, QH359-425, phylogénie, Animals, Humans, Gene, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, Synteny, génome, biology.organism_classification, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, mutation, Sequence Alignment, 030217 neurology & neurosurgery
Abstract: Background Based on the observation of an increased number of paralogous genes in teleost fishes compared with other vertebrates and on the conserved synteny between duplicated copies, it has been shown that a whole genome duplication (WGD) occurred during the evolution of Actinopterygian fish. Comparative phylogenetic dating of this duplication event suggests that it occurred early on, specifically in teleosts. It has been proposed that this event might have facilitated the evolutionary radiation and the phenotypic diversification of the teleost fish, notably by allowing the sub- or neo-functionalization of many duplicated genes. Results In this paper, we studied in a wide range of Actinopterygians the duplication and fate of the androgen receptor (AR, NR3C4), a nuclear receptor known to play a key role in sex-determination in vertebrates. The pattern of AR gene duplication is consistent with an early WGD event: it has been duplicated into two genes AR-A and AR-B after the split of the Acipenseriformes from the lineage leading to teleost fish but before the divergence of Osteoglossiformes. Genomic and syntenic analyses in addition to lack of PCR amplification show that one of the duplicated copies, AR-B, was lost in several basal Clupeocephala such as Cypriniformes (including the model species zebrafish), Siluriformes, Characiformes and Salmoniformes. Interestingly, we also found that, in basal teleost fish (Osteoglossiformes and Anguilliformes), the two copies remain very similar, whereas, specifically in Percomorphs, one of the copies, AR-B, has accumulated substitutions in both the ligand binding domain (LBD) and the DNA binding domain (DBD). Conclusion The comparison of the mutations present in these divergent AR-B with those known in human to be implicated in complete, partial or mild androgen insensitivity syndrome suggests that the existence of two distinct AR duplicates may be correlated to specific functional differences that may be connected to the well-known plasticity of sex determination in fish. This suggests that three specific events have shaped the present diversity of ARs in Actinopterygians: (i) early WGD, (ii) parallel loss of one duplicate in several lineages and (iii) putative neofunctionalization of the same duplicate in percomorphs, which occurred a long time after the WGD.
Published: 2008

46. Characterization of early molecular sex differentiation in rainbow trout, Oncorhynchus mykiss

Author: Daniel Baron, Gwenaëlle Randuineau, Chantal Cauty, Denise Vizziano, and Yann Guiguen
Subjects: Male, endocrine system, medicine.medical_specialty, animal structures, Sex Differentiation, medicine.drug_class, Somatic cell, Ovary, In situ hybridization, Aromatase, Internal medicine, medicine, Animals, Humans, Gonads, In Situ Hybridization, Sexual differentiation, biology, urogenital system, Gene Expression Profiling, Gene Expression Regulation, Developmental, biology.organism_classification, Androgen, Sexual dimorphism, Trout, Endocrinology, medicine.anatomical_structure, Oncorhynchus mykiss, Rainbow trout, Female, Developmental Biology
Abstract: Early differentiation in rainbow trout gonads was investigated by expression profiling and in situ hybridization (ISH). Expression of cyp19a1 and fst in females and sox9a1 in males were sexually dimorphic between 32 to 35 days post-fertilization (dpf). After 35 dpf, the differentiation proceeded with sexually dimorphic profiles for sox9a2, dmrt1, cyp11b2.1, amh in males and foxl2a, foxl2b, hsd3b1, inha in females. cyp17a1, cyp11a1, star, nr5a1b increased only after 40 dpf in both sexes with a slightly higher expression in females. cyp19a1 expression was localized in a cluster of somatic cells in the ventral side of female gonads, and sox9a2 and amh in somatic cells surrounding the germ cells, at 28 dpf and thereafter, both in male and female gonads. cyp11b2.1, cyp17a1, and cyp11a1 expressions were only detected in scattered somatic cells in males after 46 dpf. This confirms the early implication of cyp19a1 in trout ovarian differentiation and suggests that early testicular differentiation does not need androgen production.
Published: 2007

47. Large-scale temporal gene expression profiling during gonadal differentiation and early gametogenesis in rainbow trout

Author: Daniel, Baron, Remi, Houlgatte, Alexis, Fostier, and Yann, Guiguen
Subjects: Male, Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, Oncorhynchus mykiss, Ovary, Testis, Animals, Cluster Analysis, Female, Gametogenesis
Abstract: The overall understanding of the sex differentiation cascade in vertebrates is still growing slowly, probably because of the variety of vertebrate models used and the number of molecular players yet to be discovered. Finding conserved mechanisms among vertebrates should provide a better view of the key factors involved in this process. To this end, we used real-time reverse transcription-polymerase chain reaction to produce a temporal map of fluctuations in mRNA expression of 102 genes during sex differentiation and early gametogenesis in the rainbow trout (Oncorhynchus mykiss). We used these 102 temporal gene expression patterns as a basis for a hierarchical clustering analysis to find characteristic clusters of coexpressed genes. Analysis of some of these gene clusters suggested a conserved overall expression profile between the sex differentiation cascade in fish and mammals. Among these conserved molecular mechanisms, sox9, dmrt1, amh, nr5a1, nr0b1, igf1, and igf1ra are, for instance, characterized as early expressed genes involved in trout testicular differentiation as it is known or suggested in mammals. On the contrary, foxl2, fst, and lhr are characterized as early expressed genes during trout ovarian differentiation, as also found in mammals. Apart from this high conservation, our analysis suggests some potential new players, such as the fshb subunit gene, which is detected here for the first time, to our knowledge, in the female differentiating gonad of a vertebrate species and displays a specific overexpression that coincides in timing with the occurrence of first oocyte meioses, or the pax2 gene, which displays an early and testis-specific expression profile.
Published: 2005

48. Foxl2 gene and the development of the ovary : a story about goat, mouse, fish and woman

Author: Daniel, Baron, Frank, Batista, Stéphane, Chaffaux, Julie, Cocquet, Corinne, Cotinot, Edmond, Cribiu, Elfride, De Baere, Elfride, De Baeree, Yann, Guiguen, Francis, Jaubert, Eric, Pailhoux, Maëlle, Pannetier, Daniel, Vaiman, Bernard, Vigier, Reiner, Veitia, Marc, Fellous, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), Institut National de la Recherche Agronomique (INRA), Génomique et épigénétique des pathologies placentaires (Inserm U709), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5), Unité de recherche Génétique Biochimique et Cytogénétique (LGBC), Biologie du développement et reproduction (BDR), Centre National de la Recherche Scientifique (CNRS)-École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and Universiteit Gent = Ghent University [Belgium] (UGENT)
Subjects: Forkhead Box Protein L2, Male, Embryology, Transcription, Genetic, Disorders of Sex Development, Medicine (miscellaneous), comparative genomics, 010501 environmental sciences, medicine.disease_cause, 01 natural sciences, FOXL2 Gene, Mice, 0302 clinical medicine, Medicine and Health Sciences, Disorders of sex development, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS, Regulation of gene expression, Genetics, 0303 health sciences, Mutation, Goats, Fishes, Gene Expression Regulation, Developmental, Forkhead Transcription Factors, DNA-Binding Proteins, Testis determining factor, medicine.anatomical_structure, Forkhead box L2, DIFFERENTIATION, Female, EXPRESSION, FOXL2, Ovary, Biology, 03 medical and health sciences, Species Specificity, SRY, medicine, Animals, Humans, Gene, 0105 earth and related environmental sciences, 030304 developmental biology, TRANSCRIPTION FACTOR FOXL2, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Sex Determination Processes, medicine.disease, Reproductive Medicine, BPES, PIS, Animal Science and Zoology, ovary, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, 030217 neurology & neurosurgery, Developmental Biology, Food Science, Transcription Factors
Abstract: International audience; In this review, we describe recent results concerning the genetics of sex determination in mammals. Particularly, we developed the study of the FOXL2 gene and its implication in genetic anomalies in goats (PIS mutation) and humans (BPES). We present the expression of FOXL2 in the ovaries of different species.
Published: 2005

49. Androgen and estrogen treatments alter steady state messengers RNA (mRNA) levels of testicular steroidogenic enzymes in the Rainbow trout, Oncorhynchus mykiss

Author: Bernard Breton, Alexis Fostier, Yann Guiguen, Daniel Baron, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), and Institut National de la Recherche Agronomique (INRA)
Subjects: Male, endocrine system, medicine.medical_specialty, Time Factors, medicine.drug_class, 010501 environmental sciences, Testicle, Biology, 01 natural sciences, 03 medical and health sciences, Follicle-stimulating hormone, Internal medicine, Gene expression, Testis, Genetics, medicine, Animals, RNA, Messenger, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0105 earth and related environmental sciences, 0303 health sciences, Cholesterol side-chain cleavage enzyme, Estrogens, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Cell Biology, Androgen, biology.organism_classification, 3. Good health, Enzymes, Trout, Endocrinology, medicine.anatomical_structure, Estrogen, Oncorhynchus mykiss, Androgens, Rainbow trout, Steroids, Follicle Stimulating Hormone, Developmental Biology
Abstract: Recent investigations have shown that estrogens have profound inhibitory effects on steroidogenic enzyme gene expressions before and after testicular differentiation in the rainbow trout, Oncorhynchus mykiss. This present study bring new data on juvenile rainbow trout treated with estrogens and androgens. Following a 8 days oral treatment of juvenile male with 17alpha-ethynyl-estradiol (EE2, 20 mg/kg diet) or 11beta-hydroxyandrostenedione (11betaOHDelta4, 10 mg/kg diet), we observed a fast and marked decrease of steady-state mRNA levels for 3betaHSD, P450scc, P450c17, and P450c11 enzymes in the testis. After completion of these treatments, mRNA levels of these enzymes remained low in EE2 treated males whereas in 11betaOHDelta4 treated males they recovered their initial levels in 8 days. This demonstrate that both androgen and estrogen treatments have profound effects on testicular steroidogenesis by decreasing steroid enzymes steady-state mRNA. After in vitro incubation of testicular explants with 17beta-estradiol (E2, 600 ng/ml of medium), we also observed a decrease of mRNA levels for 3betaHSD and P450c11. This suggest that estrogens effects could be triggered, at least to some extend, directly on the testis. We also investigated the hypothesis of a negative feedback of steroids on follicle stimulating hormone (FSH) secretion, but FSH plasmatic levels in treated fish did not showed any significant decrease. This demonstrate that FSH is not implied in this steroids inhibition of steroidogenic enzymes gene expression.
Published: 2005

50. An evolutionary and functional analysis of FoxL2 in rainbow trout gonad differentiation

Author: Daniel Baron, Marc Fellous, Reiner A. Veitia, Xuhua Xia, Yann Guiguen, Julie Cocquet, Station commune de Recherches en Ichtyophysiologie, Biodiversité et Environnement (SCRIBE), and Institut National de la Recherche Agronomique (INRA)
Subjects: Male, Gonad, Sex Differentiation, medicine.drug_class, Evolution, Molecular, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Aromatase, medicine, Animals, Gonads, Molecular Biology, Transcription factor, Gene, ComputingMilieux_MISCELLANEOUS, Phylogeny, 030304 developmental biology, Genetics, 0303 health sciences, Aromatase inhibitor, Sexual differentiation, biology, GAMETOGENESE, BIOLOGIE MOLECULAIRE, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, DNA-Binding Proteins, medicine.anatomical_structure, Estrogen, Oncorhynchus mykiss, biology.protein, Rainbow trout, Female, 030217 neurology & neurosurgery, Transcription Factors
Abstract: FOXL2 is a forkhead transcription factor involved in ovarian development and function. Here, we have studied the evolution and pattern of expression of the FOXL2 gene and its paralogs in fish. We found well conserved FoxL2 sequences (FoxL2a) and divergent genes, whose forkhead domains belonged to the class L2 and were shown to be paralogs of the FoxL2a sequences (named FoxL2b). In the rainbow trout, FoxL2a and FoxL2b were specifically expressed in the ovary, but displayed different temporal patterns of expression. FoxL2a expression correlated with the level of aromatase, the key enzyme in estrogen production, and an estrogen treatment used to feminize genetically male individuals elicited the up-regulation of both paralogs. Conversely, androgens or an aromatase inhibitor down-regulated FoxL2a and FoxL2b in females. We speculate that there is a direct link between estrogens and FoxL2 expression in fish, at least during the period where the identity of the gonad is sensitive to hormonal treatments.
Published: 2004

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