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8. Looking for a needle in a haystack: de novo phenotypic target identification reveals Hippo pathway-mediated miR-202 regulation of egg production

Author

Janati-Idrissi, Sarah, primary, de Abreu, Mariana Roza, additional, Guyomar, Cervin, additional, de Mello, Fernanda, additional, Nguyen, Thaovi, additional, Mechkouri, Nazim, additional, Gay, Stéphanie, additional, Montfort, Jérôme, additional, Gonzalez, Anne Alicia, additional, Abbasi, Marzieh, additional, Bugeon, Jérôme, additional, Thermes, Violette, additional, Seitz, Hervé, additional, and Bobe, Julien, additional

Published
2023
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13. Gene expression profiling of colon cancer by DNA microarrays and correlation with histoclinical parameters

Author

Bertucci, François, Salas, Sébastien, Eysteries, Séverine, Nasser, Valéry, Finetti, Pascal, Ginestier, Christophe, Charafe-Jauffret, Emmanuelle, Loriod, Béatrice, Bachelart, Loïc, Montfort, Jérôme, Victorero, Geneviève, Viret, Frédéric, Ollendorff, Vincent, Fert, Vincent, Giovaninni, Marc, Delpero, Jean-Robert, Nguyen, Catherine, Viens, Patrice, Monges, Geneviève, Birnbaum, Daniel, and Houlgatte, Rémi

Published
2004
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14. Transcriptional rewiring, post-transcriptional regulation and neo-functionalization: how the master sex-determining gene of medaka was born

Author

Herpin, Amaury, Schmidt, Cornelia, Kneitz, Susanne, Gobe, Clara, Regensburger, Martina, Le Cam, Aurélie, Montfort, Jérôme, Adolfi, Mateus C., Lillesaar, Christina, Kraeussling, Michael, Mourot, Brigitte, Porcon, Béatrice, Pannetier, Maelle, Pailhoux, Eric, Ettwiller, Laurence, Dolle, Dirk, Guiguen, Yann, Schartl, Manfred, 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), Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Centre for Organismal Studies (COS), Department of developmental Biology, Heidelberg University, Comprehensive Cancer Center Mainfranken, Leibniz Institute of Freshwater Ecology and Inland Fisheries (IGB). DEU., 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), and University of Heidelberg

Subjects
fish, détermination du sexe, medaka, [SDV]Life Sciences [q-bio], education, gène déterminant majeur du sexe, sex determination, oryzias latipes, reproduction, poisson, gène dmrt1bY, adrianichthyidae, gonadogénèse, transcription, expression des gènes
Abstract

International audience; Transcriptional rewiring, post-transcriptional regulation and neo-functionalization: how the master sex-determining gene of medaka was born. Workshop: Paradigm shift in sex chromosome evolution.

Published
2019
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15. Oocyte-somatic cells interactions, lessons from evolution

Author

Charlier Cathy, Montfort Jérôme, Chabrol Olivier, Brisard Daphné, Nguyen Thaovi, Le Cam Aurélie, Richard-Parpaillon Laurent, Moreews François, Pontarotti Pierre, Uzbekova Svetlana, Chesnel Franck, and Bobe Julien

Subjects
Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract

Abstract Background Despite the known importance of somatic cells for oocyte developmental competence acquisition, the overall mechanisms underlying the acquisition of full developmental competence are far from being understood, especially in non-mammalian species. The present work aimed at identifying key molecular signals from somatic origin that would be shared by vertebrates. Results Using a parallel transcriptomic analysis in 4 vertebrate species - a teleost fish, an amphibian, and two mammals - at similar key steps of developmental competence acquisition, we identified a large number of species-specific differentially expressed genes and a surprisingly high number of orthologous genes exhibiting similar expression profiles in the 3 tetrapods and in the 4 vertebrates. Among the evolutionary conserved players participating in developmental competence acquisition are genes involved in key processes such as cellular energy metabolism, cell-to-cell communications, and meiosis control. In addition, we report many novel molecular actors from somatic origin that have never been studied in the vertebrate ovary. Interestingly, a significant number of these new players actively participate in Drosophila oogenesis. Conclusions Our study provides a comprehensive overview of evolutionary-conserved mechanisms from somatic origin participating in oocyte developmental competence acquisition in 4 vertebrates. Together our results indicate that despite major differences in ovarian follicular structure, some of the key players from somatic origin involved in oocyte developmental competence acquisition would be shared, not only by vertebrates, but also by metazoans. The conservation of these mechanisms during vertebrate evolution further emphasizes the important contribution of the somatic compartment to oocyte quality and paves the way for future investigations aiming at better understanding what makes a good egg.

Published
2012
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16. MiR-202 controls female fecundity by regulating medaka oogenesis

Author

Gay, Stéphanie, Bugeon, Jérôme, Bouchareb, Amine, Henry, Laure, Delahaye, Clara, Legeai, Fabrice, Montfort, Jérôme, Le Cam, Aurélie, Siegel, Anne, Bobe, Julien, Thermes, Violette, 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), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES), Centre National de la Recherche Scientifique (CNRS), 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), 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 Bretagne Sud (UBS)-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)-Institut National de Recherche en Informatique et en Automatique (Inria)-École normale supérieure - Rennes (ENS Rennes)-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-CentraleSupélec-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Université de Bretagne Sud (UBS)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-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 Rennes (UNIV-RENNES)-École normale supérieure - Rennes (ENS Rennes)-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT), Dynamics, Logics and Inference for biological Systems and Sequences (Dyliss), ANR-16-COFA-0004, Agence Nationale de la Recherche, ANR-11-INBS-0014,TEFOR,Transgenèse pour les Etudes Fonctionnelles sur les Organismes modèles(2011), ANR-16-COFA-0004,AquaCrispr,Optimization of the CRISPR/Cas9 knock-in technology and application in salmon and trout(2016), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Rennes (UR), Institut National de la Recherche Agronomique (INRA)-Université de Rennes (UR)-AGROCAMPUS OUEST, Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-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)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-IMT Atlantique (IMT Atlantique), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université de Bretagne Sud (UBS)-École normale supérieure - Rennes (ENS Rennes)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-IMT Atlantique (IMT Atlantique), AGROCAMPUS OUEST-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Recherche Agronomique (INRA), 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)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-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)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS)-IMT Atlantique Bretagne-Pays de la Loire (IMT Atlantique), and ANR-11-INBS-0014/11-INBS-0014,TEFOR,Transgenèse pour les Etudes Fonctionnelles sur les Organismes modèles(2011)

Subjects
Male, MiR-202, Physiology, [SDV]Life Sciences [q-bio], Oryzias, Biochemistry, Epithelium, Animals, Genetically Modified, Gene Knockout Techniques, oeuf de poisson, Oogenesis, poisson, Animal Cells, Reproductive Physiology, Medicine and Health Sciences, Gene Editing, Obstetrics and Gynecology, Gene Expression Regulation, Developmental, ovogenèse, microRNAs, Ovaries, Nucleic acids, OVA, micro arn, Female, Anatomy, Cellular Types, Genital Anatomy, Research Article, expression des gènes, lcsh:QH426-470, qualité des oeufs, Urology, fecundity, oryzia latipes, reproduction, Genetics, Animals, adrianichthyidae, analyse du transcriptome, Non-coding RNA, Gonads, CRISPR/Cas9, knock out, fish, medaka, Granulosa Cells, Gene Expression Profiling, ovaire, Reproductive System, Biology and Life Sciences, Subfertility, Epithelial Cells, Cell Biology, Female Subfertility, mécanisme de régulation, Gene regulation, lcsh:Genetics, Germ Cells, Biological Tissue, Fertility, Infertility, fécondite, Oocytes, croissance folliculaire, RNA, Women's Health, Lutein Cells, ovary, Gene expression, CRISPR-Cas Systems
Abstract

Female gamete production relies on coordinated molecular and cellular processes that occur in the ovary throughout oogenesis. In fish, as in other vertebrates, these processes have been extensively studied both in terms of endocrine/paracrine regulation and protein expression and activity. The role of small non-coding RNAs in the regulation of animal reproduction remains however largely unknown and poorly investigated, despite a growing interest for the importance of miRNAs in a wide variety of biological processes. Here, we analyzed the role of miR-202, a miRNA predominantly expressed in male and female gonads in several vertebrate species. We studied its expression in the medaka ovary and generated a mutant line (using CRISPR/Cas9 genome editing) to determine its importance for reproductive success with special interest for egg production. Our results show that miR-202-5p is the most abundant mature form of the miRNA and that it is expressed in granulosa cells and in the unfertilized egg. The knock out (KO) of mir-202 gene resulted in a strong phenotype both in terms of number and quality of eggs produced. Mutant females exhibited either no egg production or produced a dramatically reduced number of eggs that could not be fertilized, ultimately leading to no reproductive success. We quantified the size distribution of the oocytes in the ovary of KO females and performed a large-scale transcriptomic analysis approach to identified dysregulated molecular pathways. Together, cellular and molecular analyses indicate that the lack of miR-202 impairs the early steps of oogenesis/folliculogenesis and decreases the number of large (i.e. vitellogenic) follicles, ultimately leading to dramatically reduced female fecundity. This study sheds new light on the regulatory mechanisms that control the early steps of follicular development, including possible targets of miR-202-5p, and provides the first in vivo functional evidence that a gonad-predominant microRNA may have a major role in female reproduction., Author summary The role of small non-coding RNAs in the regulation of animal reproduction remains poorly investigated, despite a growing interest for the importance of miRNAs in a wide variety of biological processes. Here, we analyzed the role of miR-202, a miRNA predominantly expressed in gonads in vertebrate. We studied its expression in the medaka ovary and knocked out the mir-202 gene to study its importance for reproductive success. We showed that the lack of miR-202 results in the sterility of both females and males. In particular, it led to a drastic reduction of both the number and the quality of eggs produced by females. Mutant females exhibited either no egg production or produced a drastically reduced number of eggs that could not be fertilized, ultimately leading to no reproductive success. Quantitative histological and molecular analyses indicated that mir-202 KO impairs oocyte development and is also associated with the dysregulation of many genes that are critical for reproduction. This study sheds new light on the regulatory mechanisms that control oogenesis, including possible targets of miR-202-5p, and provides the first in vivo functional evidence that a gonad-predominant microRNA may have a major role in female reproduction.

Published
2018
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17. MiR-202 regulates female fecundity by controlling early follicle development in medaka ovary

Author

Gay, Stéphanie, Bugeon, Jérôme, Bouchareb, Amine, Henry, Laure, DELAHAYE, Clara, Montfort, Jérôme, Le Cam, Aurélie, Bobe, Julien, Thermes, Violette, 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 ), Instituto Nacional de Pesquisas da Amazônia (INPA). Manaus, BRA., and Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Recherche Agronomique (INRA)

Subjects
fish, medaka, qualité des oeufs, fecundity, [SDV]Life Sciences [q-bio], fungi, education, analyse d'image, ovogenèse, humanities, oryzias latipes, reproduction, oeuf de poisson, poisson, fécondite, adrianichthyidae, fluorescence, analyse du transcriptome, CRISPR/Cas9, health care economics and organizations, reproductive and urinary physiology, miRNA, expression des gènes
Abstract

MiR-202 regulates female fecundity by controlling early follicle development in medaka ovary. 11. International Symposium on Reproductive Physiology of Fish

Published
2018

18. A transcriptomic comparison suggests an influence of the domestication process on egg quality in Eurasian Perch (Perca fluviatilis)

Author

Rocha de Almeida, Taina, Alix, M., Schaerlinger, B., Le Cam, Aurélie, Montfort, Jérôme, Toomey, L., Bobe, Julien, Chardard, D., Fontaine, Pascal, Unité de Recherches Animal et Fonctionnalités des Produits Animaux (URAFPA), Institut National de la Recherche Agronomique (INRA)-Université de Lorraine (UL), 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-13-BSV7-0015 - CNPq (233389/2014-8) - MESR, CNPq (233389/2014-8) - MESR, Instituto Nacional de Pesquisas da Amazônia (INPA). Manaus, BRA., ANR-13-BSV7-0015,Maternal Legacy,Portait moléculaire d'un oeuf de poisson de bonne qualité(2013), Université de Lorraine (UL)-Institut National de la Recherche Agronomique (INRA), Unité de Recherches Animal et Fonctionnalités des Produits Animaux ( URAFPA ), Institut National de la Recherche Agronomique ( INRA ) -Université de Lorraine ( UL ), 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
fish, perca fluviatilis, [ SDV ] Life Sciences [q-bio], qualité des oeufs, [SDV]Life Sciences [q-bio], education, physiologie de la reproduction, humanities, reproduction, perch, oeuf de poisson, domestication, poisson, perche, percidae, arn maternel, health care economics and organizations, plant domestication
Abstract

A transcriptomic comparison suggests an influence of the domestication process on egg quality in Eurasian Perch (Perca fluviatilis). 11. International Symposium on Reproductive Physiology of Fish

Published
2018

19. Domestication may affect the maternal mRNA profile in unfertilized eggs, potentially impacting the embryonic development of Eurasian perch (Perca fluviatilis)

Author

Rocha de Almeida, Tainá, primary, Alix, Maud, additional, Le Cam, Aurélie, additional, Klopp, Christophe, additional, Montfort, Jérôme, additional, Toomey, Lola, additional, Ledoré, Yannick, additional, Bobe, Julien, additional, Chardard, Dominique, additional, Schaerlinger, Bérénice, additional, and Fontaine, Pascal, additional

Published
2019
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20. Expression profiling of rainbow trout testis development identifies evolutionary conserved genes involved in spermatogenesis

Author

Esquerré Diane, Ricordel Marie-Jo, Montfort Jérôme, Goupil Anne-Sophie, Lareyre Jean-Jacques, Rolland Antoine D, Hugot Karine, Houlgatte Rémi, Chalmel Fréderic, and Le Gac Florence

Subjects
Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract

Abstract Background Spermatogenesis is a late developmental process that involves a coordinated expression program in germ cells and a permanent communication between the testicular somatic cells and the germ-line. Current knowledge regarding molecular factors driving male germ cell proliferation and differentiation in vertebrates is still limited and mainly based on existing data from rodents and human. Fish with a marked reproductive cycle and a germ cell development in synchronous cysts have proven to be choice models to study precise stages of the spermatogenetic development and the germ cell-somatic cell communication network. In this study we used 9K cDNA microarrays to investigate the expression profiles underlying testis maturation during the male reproductive cycle of the trout, Oncorhynchus mykiss. Results Using total testis samples at various developmental stages and isolated spermatogonia, spermatocytes and spermatids, 3379 differentially expressed trout cDNAs were identified and their gene activation or repression patterns throughout the reproductive cycle were reported. We also performed a tissue-profiling analysis and highlighted many genes for which expression signals were restricted to the testes or gonads from both sexes. The search for orthologous genes in genome-sequenced fish species and the use of their mammalian orthologs allowed us to provide accurate annotations for trout cDNAs. The analysis of the GeneOntology terms therefore validated and broadened our interpretation of expression clusters by highlighting enriched functions that are consistent with known sequential events during male gametogenesis. Furthermore, we compared expression profiles of trout and mouse orthologs and identified a complement of genes for which expression during spermatogenesis was maintained throughout evolution. Conclusion A comprehensive study of gene expression and associated functions during testis maturation and germ cell differentiation in the rainbow trout is presented. The study identifies new pathways involved during spermatogonia self-renewal or rapid proliferation, meiosis and gamete differentiation, in fish and potentially in all vertebrates. It also provides the necessary basis to further investigate the hormonal and molecular networks that trigger puberty and annual testicular recrudescence in seasonally breeding species.

Published
2009
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21. Androgen-induced masculinization in rainbow trout results in a marked dysregulation of early gonadal gene expression profiles

Author

Houlgatte Rémi, Montfort Jérôme, Baron Daniel, Fostier Alexis, and Guiguen Yann

Subjects
Biotechnology, TP248.13-248.65, Genetics, QH426-470
Abstract

Abstract Background Fish gonadal sex differentiation is affected by sex steroids treatments providing an efficient strategy to control the sexual phenotype of fish for aquaculture purposes. However, the biological effects of such treatments are poorly understood. The aim of this study was to identify the main effects of an androgen masculinizing treatment (11β-hydroxyandrostenedione, 11βOHΔ4, 10 mg/kg of food for 3 months) on gonadal gene expression profiles of an all-female genetic population of trout. To characterize the most important molecular features of this process, we used a large scale gene expression profiling approach using rainbow trout DNA microarrays combined with a detailed gene ontology (GO) analysis. Results 2,474 genes were characterized as up-regulated or down-regulated in trout female gonads masculinized by androgen in comparison with control male or female gonads from untreated all-male and all-female genetic populations. These genes were classified in 13 k-means clusters of temporally correlated expression profiles. Gene ontology (GO) data mining revealed that androgen treatment triggers a marked down-regulation of genes potentially involved in early oogenesis processes (GO 'mitotic cell cycle', 'nucleolus'), an up-regulation of the translation machinery (GO 'ribosome') along with a down-regulation of proteolysis (GO 'proteolysis', 'peptidase' and 'metallopeptidase activity'). Genes considered as muscle fibres markers (GO 'muscle contraction') and genes annotated as structural constituents of the extracellular matrix (GO 'extracellular matrix') or related to meiosis (GO 'chromosome' and 'meiosis') were found significantly enriched in the two clusters of genes specifically up-regulated in androgen-treated female gonads. GO annotations 'Sex differentiation' and 'steroid biosynthesis' were enriched in a cluster of genes with high expression levels only in control males. Interestingly none of these genes were stimulated by the masculinizing androgen treatment. Conclusion This study provides evidence that androgen masculinization results in a marked dysregulation of early gene expression profiles when compared to natural testicular or ovarian differentiation. Based on these results we suggest that, in our experimental conditions, androgen masculinization proceeds mainly through an early inhibition of female development.

Published
2007
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22. Muscle growth and remodeling in trout: insights from transcriptomics

Author

Rescan, Pierre-Yves, Montfort, Jérôme, Le Cam, Aurélie, Ralliere, Cécile, Gabillard, Jean-Charles, Fautrel, Alain, 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 ), and Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )

Subjects
salmonidae, poisson, muscle, [SDV]Life Sciences [q-bio], croissance animale, [INFO]Computer Science [cs], truite, ComputingMilieux_MISCELLANEOUS, étude transcriptomique
Abstract

International audience

Published
2018

23. MicroRNA-202 (miR-202) controls female fecundity by regulating medaka oogenesis

Author

Gay, Stéphanie, Bugeon, Jérôme, Bouchareb, Amine, Henry, Laure, Montfort, Jérôme, Le Cam, Aurélie, Bobe, Julien, and Thermes, Violette

Subjects
medaka, fish, fertility, ovary, gonads, microRNAs, quantitative image analyses, CRISPR/cas9, microarray, ovaire, analyse d'image, mécanisme de régulation, oryzias latipes, gonade, reproduction, fertilité, reproduction femelle, poisson, micro arn, adrianichthyidae, analyse du transcriptome, expression des gènes
Abstract

Female gamete production relies on coordinated molecular and cellular processes that occur in the ovary throughout oogenesis. In fish, as in other vertebrates, these processes have been extensively studied both in terms of endocrine/paracrine regulation and protein expression and activity. The role of small non-coding RNAs in the regulation of animal reproduction remains however largely unknown and poorly investigated, despite a growing interest for the importance of miRNAs in a wide variety of biological processes. Here, we analyzed the role of miR-202, a miRNA predominantly expressed in male and female gonads in several vertebrate species. We studied its expression in the medaka ovary and generated a mutant line (using CRISPR/Cas9 genome engineering) to determine its importance for reproductive success with special interest for egg production. Our results show that miR-202-5p is the biologically active form of the miRNA and that it is expressed in granulosa cells and in the unfertilized egg. The knock out (KO) of miR-202 resulted in a strong phenotype both in terms of number and quality of eggs produced. Mutant females exhibited either no egg production or produced a drastically reduced number of eggs that could not be fertilized, ultimately leading to no reproductive success. We quantified the size distribution of the oocytes in the ovary of KO females and performed a genome-wide transcriptomic analysis approach to identified dysregulated molecular pathways. Together, cellular and molecular analyses indicate that lack of miR-202 impairs the early steps of oogenesis/folliculogenesis and decreases the number of large (i.e. vitellogenic) follicles, ultimately leading to dramatically reduced female fecundity. This study sheds new light on the regulatory mechanisms that control the early steps of follicular development and provides the first in vivo functional evidence that an ovarian-predominant microRNA may have a major role in female reproduction.

Published
2018

25. Identification of novel ovarian predominant MiRNAs in medaka

Author

Bouchareb, Amine, Le Cam, Aurélie, Montfort, Jérôme, Gay, Stéphanie, Nguyen, Thuy Thao Vi, Bobe, Julien, Thermes, Violette, 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
reproduction, medaka, poisson, développement embryonnaire, [SDV]Life Sciences [q-bio], ovaire, [INFO]Computer Science [cs], adrianichthyidae, oryzias latipes, miRNA, expression des gènes
Abstract

International audience; MicroRNAs (miRNAs) are small and highly conserved non coding RNAs involved in the regulation of thousands of genes. miRNAs play important roles in both normal physiological and pathological pathways in many organisms. The involvement of miRNAs in vertebrate oogenesis remains however poorly documented. The present study aimed at getting new insight on the role of miRNAs fish oogenesis and maternal contribution to early embryonic development. Based on the assumption that ovarian-specific or ovarian-predominant genes usually play important roles in oogenesis and/or early development in vertebrates, we searched for ovarian-predominant miRNAs in the medaka (Oryzias latipes) ovary. We designed a genome-wide microarray displaying 3800 distinct miRNAs from different teleost and vertebrate species. The microarray analysis was performed using 10 different tissues of adult medaka, including the ovary. We identified twenty miRNAs predominantly expressed in the ovary that had never been described in medaka. Among them, miR-202 was previously shown to be predominantly expressed in the rainbow trout gonads. Among these novel ovarian medaka miRNAs, three had previously been identified in the mouse ovary or germline (miR-743a, miR-878 and miR487b), and sixteen (including miR-6352, miR-478, miR-1305, and miR-5581) had never been reported as ovarian-predominant miRNAs in any vertebrate species. To characterize the cognate miRNA sequences in medaka, heterologous miRNA sequences were aligned to the medaka genome and to medaka miRNA sequences predicted from existing RNA-seq data. Real time PCR was used to confirm ovarian-predominant expression of novel miRNAs and to analyze their expression profiles during oogenesis and early embryogenesis. Here we show that miR-202 is differentially expressed throughout oogenesis and is maternally-inherited in the medaka egg. This suggests a role of miR-202 during oocyte formation process and/or early embryonic development in medaka. Finally, our study led to the identification of previously uncharacterized medaka miRNAs predominantly expressed in the ovary, and should contribute to a further understanding of the roles of specific miRNAs in female gamete formation in this model species.

Published
2015

26. Nourrir les alevins de truite pendant une période courte avec un aliment végétal améliore son utilisation ultérieure

Author

Balasubramanian, Mukundh, Médale, Françoise, Kaushik, Sadasivam, Quillet, Edwige, Dupont-Nivet, Mathilde, Le Cam, Aurélie, Montfort, Jérôme, Panserat, Stéphane, Geurden, Inge, Nutrition, Aquaculture et Génomique (NUAGE), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université Sciences et Technologies - Bordeaux 1-Institut National de la Recherche Agronomique (INRA), Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Institut National de la Recherche Agronomique (INRA), 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), Institut National de la Recherche Agronomique (INRA)-Université Sciences et Technologies - Bordeaux 1-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), 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 ), and Institut Technique de l'Aviculture et des Elevages de Petits Animaux (ITAVI). FRA.

Subjects
salmonidae, [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], poisson, alevin, aliment d'origine végétale, alimentation animale, truite, nutrition animale, Autre (Sciences du Vivant), expression des gènes
Abstract

Session Nutrition; absent

Published
2014

28. Identification of differentially expressed miRNAs and their potential targets during fish ovarian development

Author

Juanchich, Amélie, Le Cam, Aurélie, Montfort, Jérôme, Guiguen, Yann, Bobe, Julien, 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), Institut National de la Recherche Agronomique (INRA) PHASE grant, and Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )

Subjects
microRNA, [SDV.BA]Life Sciences [q-bio]/Animal biology, Gene Expression Profiling, Ovary, Gene Expression Regulation, Developmental, rainbow trout, MicroRNAs, metazoans, Oogenesis, Oncorhynchus mykiss, Oocytes, Animals, Female, microarray
Abstract

Supplementary data (sur le site de la revue) : 105361SuppFigS1V2 - 105361SuppTableS1 - 105361SuppTableS2 - 105361SuppTableS3; Oogenesis is a complex process requiring the coordinated sequential expression of specific genes and ultimately leading to the release of the female gamete from the ovary. In the present study we aimed to investigate the contribution of miRNAs to the regulation of this key biological process in teleosts using a model in which growing oocytes develop simultaneously. Taking advantage of the strong sequence conservation of miRNAs among phylogenetically distant species, we designed a generic microarray displaying most known chordate miRNAs. It allowed us to provide an overview of the ovarian miRNome during oogenesis for the first time in any vertebrate species. We identified 13 differentially expressed miRNAs, and a differential expression of at least one miRNA was observed at each step of oogenesis. A surprisingly high differential expression of several miRNAs was observed at several stages of oogenesis and subsequently confirmed using quantitative PCR. By refining in silico prediction of target genes with gene expression data obtained within the same sample set, we provide strong evidence that miRNAs target major players of oogenesis, including genes involved in rate-limiting steps of steroidogenesis and those involved in gonadotropic control of oocyte development, as well as genes involved in ovulation, oocyte hydration, and acquisition of the ability of the oocyte to support further development once fertilized (i.e., oocyte developmental competence). Together, these observations stress the importance of miRNAs in the regulation and success of female gamete formation during oogenesis.

Published
2013
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29. FIBERFISH - Identification et caractérisation fonctionnelle de gènes régulateurs de l'hyperplasie musculaire, une composante essentielle de la croissance et de la texture du muscle chez les poissons

Author

Rescan, Pierre-Yves, Ralliere, Cécile, Montfort, Jérôme, Lebret, Véronique, Le Cam, Aurélie, Cousin, Xavier, 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), and Plateforme Génomique Santé Biogenouest®

Subjects
différenciation musculaire, poisson, expression génique, muscle, [SDV]Life Sciences [q-bio], cellule souche, croissance animale, [INFO]Computer Science [cs], myotome, hyperplasie, myogénèse, texture, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2010

31. Toxicogénomique des perturbateurs endocriniens de la reproduction chez la truite arc-en-ciel

Author

Baron, Daniel, Bobe, Julien, Fostier, Alexis, Guiguen, Yann, Haon-Lasportes, Emilie, Le Gac, Florence, Montfort, Jérôme, Rime, Hélène, and Monod, Gilles

Subjects
salmonidae, perturbateur endocrinien, oncorhynchus mykiss, poisson, puce à adn, reproduction animale, génomique fonctionnelle, toxicogénomique, Ecotoxicologie, Ecotoxicology, transcriptome, truite arc en ciel, expression des gènes
Published
2008

32. The rainbow trout genome provides novel insights into evolution after whole-genome duplication in vertebrates

Author

Berthelot, Camille, primary, Brunet, Frédéric, additional, Chalopin, Domitille, additional, Juanchich, Amélie, additional, Bernard, Maria, additional, Noël, Benjamin, additional, Bento, Pascal, additional, Da Silva, Corinne, additional, Labadie, Karine, additional, Alberti, Adriana, additional, Aury, Jean-Marc, additional, Louis, Alexandra, additional, Dehais, Patrice, additional, Bardou, Philippe, additional, Montfort, Jérôme, additional, Klopp, Christophe, additional, Cabau, Cédric, additional, Gaspin, Christine, additional, Thorgaard, Gary H., additional, Boussaha, Mekki, additional, Quillet, Edwige, additional, Guyomard, René, additional, Galiana, Delphine, additional, Bobe, Julien, additional, Volff, Jean-Nicolas, additional, Genêt, Carine, additional, Wincker, Patrick, additional, Jaillon, Olivier, additional, Crollius, Hugues Roest, additional, and Guiguen, Yann, additional

Published
2014
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34. Characterization of rainbow trout gonad, brain and gill deep cDNA repertoires using a Roche 454-Titanium sequencing approach

Author

Le Cam, Aurélie, primary, Bobe, Julien, additional, Bouchez, Olivier, additional, Cabau, Cédric, additional, Kah, Olivier, additional, Klopp, Christophe, additional, Lareyre, Jean-Jacques, additional, Le Guen, Isabelle, additional, Lluch, Jérôme, additional, Montfort, Jérôme, additional, Moreews, Francois, additional, Nicol, Barbara, additional, Prunet, Patrick, additional, Rescan, Pierre-Yves, additional, Servili, Arianna, additional, and Guiguen, Yann, additional

Published
2012
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37. Inhibition of zygotic DNA repair: transcriptome analysis of the offspring in trout (Oncorhynchus mykiss)

Author

Biologia Celular, Fernández Díez, Cristina, González Rojo, Silvia, Montfort, Jérôme, Le Cam, Aurélie, Bobe, Julien, Robles Rodríguez, Vanesa, Pérez Cerezales, Serafín, Herráez Ortega, María Paz, Biologia Celular, Fernández Díez, Cristina, González Rojo, Silvia, Montfort, Jérôme, Le Cam, Aurélie, Bobe, Julien, Robles Rodríguez, Vanesa, Pérez Cerezales, Serafín, and Herráez Ortega, María Paz

Abstract

[EN] Zygotic repair of the paternal genome is a key event after fertilization. Spermatozoa accumulate DNA strand breaks during spermatogenesis and can suffer additional damage by different factors, including cryopreservation. Fertilization with DNA-damaged spermatozoa (DDS) is considered to promote implantation failures and abortions, but also long-term effects on the progeny that could be related with a defective repair. Base excision repair (BER) pathway is considered the most active in zygotic DNA repair, but healthy oocytes contain enzymes for all repairing pathways. In this study, the effects of the inhibition of the BER pathway in the zygote were analyzed on the progeny obtained after fertilization with differentially DDS. Massive gene expression (GE; 61 657 unique probes) was analyzed after hatching using microarrays. Trout oocytes are easily fertilized with DDS and the high prolificacy allows live progeny to be obtained even with a high rate of abortions. Nevertheless, the zygotic inhibition of Poly (ADP-ribose) polymerase, upstream of BER pathway, resulted in 810 differentially expressed genes (DEGs) after hatching. DEGs are related with DNA repair, apoptosis, telomere maintenance, or growth and development, revealing a scenario of impaired DNA damage signalization and repair. Downregulation of the apoptotic cascade was noticed, suggesting a selection of embryos tolerant to residual DNA damage during embryo development. Our results reveal changes in the progeny from defective repairing zygotes including higher malformations rate, weight gain, longer telomeres, and lower caspase 3/7 activity, whose long-term consequences should be analyzed in depth

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