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4. Human Luteinizing Hormone and Chorionic Gonadotropin Display Biased Agonism at the LH and LH/CG Receptors

Author

Riccetti, Laura, Yvinec, Romain, Klett, Danièle, Gallay, Nathalie, Combarnous, Yves, Reiter, Eric, Simoni, Manuela, Casarini, Livio, and Ayoub, Mohammed Akli

Subjects
Quantitative Biology - Molecular Networks
Abstract

Human luteinizing hormone (LH) and chorionic gonadotropin (hCG) have been considered biologically equivalent because of their structural similarities and their binding to the same receptor; the LH/CGR. However, accumulating evidence suggest that LH/CGR differentially responds to the two hormones triggering differential intracellular signaling and steroidogenesis. The mechanistic basis of such differential responses remains mostly unknown. Here, we compared the abilities of recombinant rhLH and rhCG to elicit cAMP, \beta-arrestin 2 activation, and steroidogenesis in HEK293 cells and mouse Leydig tumor cells (mLTC-1). For this, BRET and FRET technologies were used allowing quantitative analyses of hormone activities in real-time and in living cells. Our data indicate that rhLH and rhCG differentially promote cell responses mediated by LH/CGR revealing interesting divergences in their potencies, efficacies and kinetics: rhCG was more potent than rhLH in both HEK293 and mLTC-1 cells. Interestingly, partial effects of rhLH were found on \beta-arrestin recruitment and on progesterone production compared to rhCG. Such a link was further supported by knockdown experiments. These pharmacological differences demonstrate that rhLH and rhCG act as natural biased agonists. The discovery of novel mechanisms associated with gonadotropin-specific action may ultimately help improve and personalize assisted reproduction technologies

Published
2018
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9. Highly-Sensitive In Vitro Bioassays for FSH, TSH, PTH, Kp, and OT in Addition to LH in Mouse Leydig Tumor Cell

Author

Klett, Danièle, primary, Pellissier, Lucie, additional, Lomet, Didier, additional, Derouin-Tochon, Flavie, additional, Robert, Vincent, additional, Nguyen, Thi Mong Diep, additional, Duittoz, Anne, additional, Reiter, Eric, additional, Locatelli, Yann, additional, Dupont, Joëlle, additional, Dardente, Hugues, additional, Jean-Alphonse, Frédéric, additional, and Combarnous, Yves, additional

Published
2023
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21. Receptors | Thyroid-Stimulating Hormone/Luteinizing Hormone/Follicle-Stimulating Hormone Receptors

Author

Combarnous, Yves, Crépieux, Pascale, Reiter, Eric, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Dynamiques de populations multi-échelles pour des systèmes physiologiques (MUSCA), Inria Saclay - Ile de France, Institut National de Recherche en Informatique et en Automatique (Inria)-Institut National de Recherche en Informatique et en Automatique (Inria)-Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Mathématiques et Informatique Appliquées du Génome à l'Environnement [Jouy-En-Josas] (MaIAGE), 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 Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Mathématiques et Informatique Appliquées du Génome à l'Environnement [Jouy-En-Josas] (MaIAGE), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Mathématiques et Informatique Appliquées du Génome à l'Environnement [Jouy-En-Josas] (MaIAGE)

Subjects
LH, Gonadotropin, Thyroid, endocrine system, 0303 health sciences, TSH, [SDV]Life Sciences [q-bio], Lutropin, Ovary, Thyrotropin, Follitropin, TSHR, 03 medical and health sciences, Thyrotropin receptor autoantibodies TRAbs, 0302 clinical medicine, FSHR, 030220 oncology & carcinogenesis, FSH, LHCGR, Glycoprotein hormone, G protein-coupled receptor, Testes, hormones, hormone substitutes, and hormone antagonists, 030304 developmental biology
Abstract

International audience; The thyroid-stimulating hormone receptor (TSHR), luteinizing hormone receptor (LHCGR), and follicle-stimulating hormone receptor (FSHR) are collectively referred to as the glycoprotein hormone receptors because they bind structurally similar glycoprotein hormones. The glycoprotein hormones consist of the pituitary thyroid-stimulating hormone (thyrotropin, TSH), luteinizing hormone (lutropin, LH), and follicle-stimulating hormone (follitropin, FSH) as well as the chorionic gonadotropin (choriogonadotropin, CG) placental hormone. The glycoprotein hormones are each composed of two dissimilar subunits (α and β) that are noncovalently associated. Within a given species, the α-subunit is identical and the β-subunits are distinct but homologous. Due to the largely similar nature of LHβ and CGβ, the LHCGR binds either LH or CG. The binding specificity of LH/CG to the LHCGR, FSH to the FSHR, and TSH to the TSHR is fairly strict. However, in some cases, extremely high levels of hormone can cause activation of the inappropriate receptor. Because the LHCGR and FSHR are localized primarily to the gonads, these two receptors are also referred to as the gonadotropin receptors. The glycoprotein hormone receptors belong to the family A (rhodopsin-like) of G protein-coupled receptors (GPCRs). However, they share the relatively unique feature of having a large extracellular domain that mediates high affinity binding of the hormone. In spite of their different physiological roles, the glycoprotein hormone receptors share a similar structural organization and mechanism of action.

Published
2021
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33. Comparative structure analyses of cystine knot-containing molecules with eight aminoacyl ring including glycoprotein hormones (GPH) alpha and beta subunits and GPH-related A2 (GPA2) and B5 (GPB5) molecules

Author

Combarnous Yves, Cahoreau Claire, and Alvarez Eva

Subjects
Gynecology and obstetrics, RG1-991, Reproduction, QH471-489
Abstract

Abstract Background Cystine-knot (cys-knot) structure is found in a rather large number of secreted proteins and glycoproteins belonging to the TGFbeta and glycoprotein hormone (GPH) superfamilies, many of which are involved in endocrine control of reproduction. In these molecules, the cys-knot is formed by a disulfide (SS) bridge penetrating a ring formed by 8, 9 or 10 amino-acid residues among which four are cysteine residues forming two SS bridges. The glycoprotein hormones Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Thyroid-Stimulating Hormone (TSH) and Chorionic Gonadotropin (CG) are heterodimers consisting of non-covalently associated alpha and beta subunits that possess cys-knots with 8-amino-acyl (8aa) rings. In order to get better insight in the structural evolution of glycoprotein hormones, we examined the number and organization of SS bridges in the sequences of human 8-aa-ring cys-knot proteins having 7 (gremlins), 9 (cerberus, DAN), 10 (GPA2, GPB5, GPHα) and 12 (GPHβ) cysteine residues in their sequence. Discussion The comparison indicated that the common GPH-alpha subunit exhibits a SS bridge organization ressembling that of DAN and GPA2 but possesses a unique bridge linking an additional cysteine inside the ring to the most N-terminal cysteine residue. The specific GPHbeta subunits also exhibit a SS bridge organization close to that of DAN but it has two additional C-terminal cysteine residues which are involved in the formation of the "seat belt" fastened by a SS "buckle" that ensures the stability of the heterodimeric structure of GPHs. GPA2 and GPB5 exhibit no cys residue potentially involved in interchain SS bridge and GPB5 does not possess a sequence homologous to that of the seatbelt in GPH β-subunits. GPA2 and GPB5 are thus not expected to form a stable heterodimer at low concentration in circulation. Summary The 8-aa cys-knot proteins GPA2 and GPB5 are expected to form a heterodimer only at concentrations above 0.1 microM: this would be consistent with a short-term paracrine role but not with an endocrine role after dilution in circulation. Consequently, GPA2 and GPB5 could exert separate endocrine roles either during development and/or during adult life of both vertebrates and invertebrates.

Published
2009
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35. Fluoxetine affects cytosolic cAMP, ATP, Ca2+ responses to forskolin, and survival of human ovarian granulosa tumor COV434 cells.

Author

Thi Mong Diep Nguyen, Klett, Danièle, and Combarnous, Yves

Subjects
GRANULOSA cell tumors, OVARIAN tumors, INTRACELLULAR calcium, FORSKOLIN, FLUOXETINE, SEROTONIN uptake inhibitors
Abstract

Fluoxetine (FLX), a selective serotonin reuptake inhibitor antidepressant, exhibits various other mechanisms of action in numerous cell types and has been shown to induce cell death in cancer cells, paving the way for its potential use in cancer therapy. The aim of this study was to determine the off-target effects of the anti-depressant drug FLX, on the human ovarian granulosa tumor COV434 cells stimulated by forskolin (FSK), by measuring the real-time kinetics of intracellular cyclic AMP (cAMP), ATP level, cytoplasmic calcium ([Ca2+]cyt) and survival of COV434 cells. We show that incubating COV434 cells with FLX (between 0.6 and 10 µM) induces a decrease in intracellular cAMP response to FSK, a drop in ATP content and stimulates cytoplasmic Ca2+ accumulation in COV434 cells. Only the highest concentrations of FLX (5-10 µM) diminished cell viability. The present report is the first to identify an action mechanism of FLX in human tumor ovarian cells COV434 cells and thus opening the way to potential use of fluoxetine as a complementary tool, in granulosa tumor treatments. [ABSTRACT FROM AUTHOR]

Published
2021
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37. Permissive effect of substimulating doses of natural and recombinant human LH and hCG but not rat, ovine, bovine, porcine, equine LHs and equine CG on subsequent MLTC-1 adenylate cyclase responsiveness to forskolin (FSK)

Author

Nguyen, Thi-Mong-Diep, Klett, Danièle, Combarnous, Yves, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), ProdInra, Archive Ouverte, Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Université François Rabelais (Tours). Tours, FRA., and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT], Leydig, hCG, hormone, cyclic AMP, forskoline, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, hLH, adenylate cyclase, forskolin
Abstract

Permissive effect of substimulating doses of natural and recombinant human LH and hCG but not rat, ovine, bovine, porcine, equine LHs and equine CG on subsequent MLTC-1 adenylate cyclase responsiveness to forskolin (FSK). 2. Journées du GdR 3606 Repro

Published
2017

38. Fast renal trapping of porcine Luteinizing Hormone (pLH) shown by 123I-scintigraphic imaging in rats explains its short circulatory half-life

Author

Locatelli Alain, Magallon Thierry, Leroux Hervé, Lecompte François, Bernard Serge, Klett Danièle, Lepape Alain, and Combarnous Yves

Subjects
Gynecology and obstetrics, RG1-991, Reproduction, QH471-489
Abstract

Abstract Background Sugar moieties of gonadotropins play no primary role in receptor binding but they strongly affect their circulatory half-life and consequently their in vivo biopotencies. In order to relate more precisely hepatic trapping of these glycoproteic hormones with their circulatory half-life, we undertook a comparative study of the distribution and elimination of porcine LH (pLH) and equine CG (eCG) which exhibit respectively a short and a long half-life. This was done first by following half-lives of pLH in piglets with hepatic portal circulation shunted or not. It was expected that such a shunt would enhance the short half-life of pLH. Subsequently, scintigraphic imaging of both 123I-pLH and 123I-eCG was performed in intact rats to compare their routes and rates of distribution and elimination. Methods Native pLH or eCG was injected to normal piglets and pLH was tested in liver-shunted anæsthetized piglet. Blood samples were recovered sequentially over one hour time and the hormone concentrations were determined by a specific ELISA method. Scintigraphic imaging of 123I-pLH and 123I-eCG was performed in rats using a OPTI-CGR gamma camera. Results In liver-shunted piglets, the half-life of pLH was found to be as short as in intact piglets (5 min). In the rat, the half-life of pLH was also found to be very short (3–6 min) and 123I-pLH was found to accumulate in high quantity in less than 10 min post injection at the level of kidneys but not in the liver. 123I-eCG didn't accumulate in any organ in the rats during the first hour, plasma concentrations of this gonadotropin being still elevated (80%) at this time. Conclusion In both the porcine and rat species, the liver is not responsible for the rapid elimination of pLH from the circulation compared to eCG. Our scintigraphic experiments suggest that the very short circulatory half-life of LH is due to rapid renal trapping.

Published
2003
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41. Comparison of natural and recombinant Glycoprotein Hormones Carbohydrate moieties by Sandwich Enzyme-linked Lectin Immuno-Assays (SELIA)

Author

Klett, Danièle, Combarnous, Yves, ProdInra, Archive Ouverte, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Université François Rabelais (Tours). Tours, FRA. Centre National de la Recherche Scientifique (CNRS), FRA., and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)

Subjects
LH, sandwich assay, glycoprotein, glycoprotéine, [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT], FSH, hormone, CG, lectin, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology
Abstract

Comparison of natural and recombinant Glycoprotein Hormones Carbohydrate moieties by Sandwich Enzyme-linked Lectin Immuno-Assays (SELIA). 2. Journées du GdR 3606 Repro

Published
2017

43. Heterogeneous hCG and hMG commercial preparations result in biased intracellular signaling but induce similar progesterone response in vitro

Author

Riccetti, Laura, primary, Klett, Daniele, additional, Ayoub, Mohammed Akli, additional, Boulo, Thomas, additional, Mariot, Julie, additional, Pignatti, Elisa, additional, Tagliavini, Simonetta, additional, Varani, Manuela, additional, Tranti, Tommaso, additional, Nicoli, Alessia, additional, Sala, Giovanni Battista La, additional, Combarnous, Yves, additional, Reiter, Eric, additional, Simoni, Manuela, additional, and Casarini, Livio, additional

Published
2017
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45. Reference preparations and bioassays for gonadotropin preparations: why do pharmacopeas ignore scientific data?

Author

Combarnous, Yves, Mariot, Julie, Jouanny, Camille, Rafert, Sandrine, Klett, Danièle, Cahoreau, Claire, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Université de Haute Bretagne ( Rennes 2 ) (UR 2). Rennes, FRA., Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), and ProdInra, Migration

Subjects
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT], ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2015

46. Communications et signalisations cellulaires (4° Éd.)

Author

COMBARNOUS Yves and COMBARNOUS Yves

Subjects
Cell phone systems, Personal communication service systems, Wireless communication systems
Abstract

Les activités des cellules d'un organisme complexe doivent être coordonnées via des régulations intégratives d'ordre supérieur (endocrines, nerveuses, immunitaires). Ces coordinations mettent en jeu des communications intercellulaires grâce à des messagers (hormones, neuromédiateurs, cytokines, phéromones) qui, en se liant à des récepteurs spécifiques de leurs cellules cibles, déclenchent, en aval, l'activation de voies de signalisation intracellulaires. Celles-ci sont largement conservées chez tous les eucaryotes multicellulaires, y compris l'espèce humaine. Cette nouvelle édition de Communications et signalisations cellulaires, entièrement remise à jour et remaniée, dresse un état des lieux des connaissances dans ce vaste domaine avec pour principal objectif non pas d'accumuler de trop nombreuses données, mais d'offrir au lecteur des éléments simples et pertinents lui permettant de situer ses molécules d'intérêt dans un cadre plus général. Pratique, didactique et toujours agrémenté d'un lexique franco-anglais des sigles, cet ouvrage s'adresse aux enseignants, chercheurs et étudiants en biologie, biochimie et médecine pour lesquels il constitue une véritable référence et un guide indispensable.

Published
2013

47. Les gonadotropines Ed. 1

Author

Combarnous, Yves, Combarnous, Yves, Combarnous, Yves, and Combarnous, Yves

Abstract

Premier ouvrage de synthèse en français consacré à la description physico-chimique, immunologique, biologique et clinique des gonadotropines, ces hormones complexes qui jouent un rôle central dans la reproduction, chez l'homme et les vertébrés. La contribution d'une trentaine des meilleurs spécialistes français offre aux chercheurs, biochimistes ou endocrinologistes, aux médecins et vétérinaires ainsi qu'aux professeurs et étudiants de 3e cycle une mise à jour des connaissances actuelles sur le sujet.

Published
2006

48. Thermal stability of Single-Chain bovine FSH with hCG-CTP linker

Author

Jouanny, Camille, Mariot, Julie, Amrane, Samir, Klett, Danièle, Cahoreau, Claire, Jegot, Gwenhael, Bourdoncle, Anne, Combarnous, Yves, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), ARN : régulations naturelle et artificielle, Université Bordeaux Segalen - Bordeaux 2-Institut Européen de Chimie et de Biologie-Institut National de la Santé et de la Recherche Médicale (INSERM), Le Studium - Loire Valley Institute for Advanced Studies. Orléans, FRA. Institut National de la Recherche Agronomique (INRA)., ProdInra, Migration, and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], [SDV.OT] Life Sciences [q-bio]/Other [q-bio.OT], ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2014

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