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9. CHARACTERIZATION OF ESTROGEN PRODUCTION AND FUNCTION IN ADULT RABBIT TESTES

Author

Dewaele, Aurélie, Dujardin, Emilie, Jammes, Hélène, Giton, Franck, Mandon-Pepin, Béatrice, Sellem, Eli, André, Marjolaine, Albina, Audrey, Pailhoux, Eric, Jolivet, Geneviève, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire - Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), IMRB - 'NeuroPsychologie Interventionnelle' [Créteil] (U955 Inserm - UPEC), Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Allice, and Pannetier, Maëlle

Subjects
[SDV.BDD.GAM] Life Sciences [q-bio]/Development Biology/Gametogenesis, endocrine system, [SDV.BDLR.RS] Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, [SDV]Life Sciences [q-bio], Estrogens, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction, [SDV] Life Sciences [q-bio], reproduction, Sperm motility, Aromatase, Testis, CYP19A1, Rabbits, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, hormones, hormone substitutes, and hormone antagonists, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

International audience; The rise of male fertility disorders these last few years appears to be linked to the increase of pollutants and other endocrine disruptors in our environment. Some of these factors can mimic or alter estrogen signaling. Therefore it is essential to better understand the role of estrogens in the male gamete production, in the aim to preserve fertility. Estrogens are steroid hormones produced by the aromatization of androgens through the enzyme aromatase, encoded by the CYP19A1 gene. Although they are usually referred as “female sex hormones”, estrogens are also produced in the adult testis. In mammalian species where CYP19A1/Aromatase expression has been studied, it is noteworthy that the localization of estrogen production varies. The rabbit is an important biomedical model and, to date, the role of estrogens in rabbit testis function is unknown. First, we identified cells expressing CYP19A1 and we showed that testicular estrogens are produced within the seminiferous tubules by germ cells. Then, testicular estrogen and androgen levels were quantified by gas chromatography/mass spectrometry (GC/MS), comparing wild-type and CYP19A1-/- rabbits generated by CRISPR/Cas9 technology. We showed that CYP19A1-/- testes were devoid of 17b-estradiol, while testosterone or androstenedione levels were not modified. As fertility was decreased in male CYP19A1-/- rabbits lacking estrogens (reduction of the number of pups per litter), we investigated several aspects, such as testis histology, DNA methylation of germ /sperm cells and sperm parameters. In particular, CYP19A1-/- males showed reduced sperm motility associated with increased flagellar defects. To highlight estrogen effects during spermatogenesis, high throughput transcriptomic studies are in progress.

Published
2021

11. DMRT1 is Required for Fetal Germ Cell Development in Rabbit Ovary

Author

Mellouk, Namya, Dujardin, Emilie, Jolivet, Geneviève, Dewaele, Aurélie, Pailhoux, Eric, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), and Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)

Subjects
[SDV]Life Sciences [q-bio], [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

International audience

Published
2021

12. DMRT1 is Required For Sex Determination in Rabbits

Author

Dujardin, Emilie, Mellouk, Namya, André, Marjolaine, Mandon-Pepin, Béatrice, Dewaele, Aurélie, Canon, Eugénie, Jolivet, Geneviève, Pailhoux, Eric, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
[SDV]Life Sciences [q-bio], [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

International audience

Published
2021

13. Single-cell RNA Transcriptome Atlas of the Developing Fetal Rabbit Ovary

Author

Mellouk, Namya, Mandon-Pepin, Béatrice, Dewaele, Aurélie, Jouneau, Luc, Dujardin, Emilie, Aubert-Frambourg, Anne, Jolivet, Geneviève, Pailhoux, Eric, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), and École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)

Subjects
[SDV]Life Sciences [q-bio], [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

International audience

Published
2021

14. Caractérisation des trajectoires de différenciation des cellules germinales au cours de la différenciation de l'ovaire fœtal chez le lapin

Author

Mellouk, Namya, Mandon-Pepin, Béatrice, Dewaele, Aurélie, Jouneau, Luc, Dujardin, Emilie, Aubert-Frambourg, Anne, Jolivet, Geneviève, Pailhoux, Eric, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), ACI PHASE INRA CAROTANR ARDIGERM ANR-20-CE14-0022, and ANR-20-CE14-0022,ARDIGERM,Acide rétinoïque dans la différenciation des cellules germinales et la méiose(2020)

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

International audience

Published
2021

15. Contributions maternelles et zygotiques à la reprogrammation des enhancers chez les embryons bovins pré-implantoires

Author

Halstead, Michelle, Laffont, Ludivine, Aubert-Frambourg, Anne, Jouneau, Luc, Canon, Eugénie, Pannetier, Maëlle, Bonnet-Garnier, Amélie, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), and API PHASE INRAE EpiNEXT

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2021

16. Dynamique de l’épigénome des cellules germinales femelles lors de l’initiation de la méiose dans l’ovaire fœtal de lapin : Analyse des modifications post-traductionnelles des histones par CUT&RUN

Author

Canon, Eugénie, Mellouk, Namya, Dewaele, Aurélie, Mandon-Pepin, Béatrice, Jouneau, Luc, Halstead, Michelle, Aubert-Frambourg, Anne, Jolivet, Geneviève, Bonnet-Garnier, Amélie, Pailhoux, Eric, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), API PHASE INRAE epiNEXTANR ARDIGERM ANR-20-CE14-0022, and ANR-20-CE14-0022,ARDIGERM,Acide rétinoïque dans la différenciation des cellules germinales et la méiose(2020)

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

National audience

Published
2021

17. Différenciation et reprogrammation épigénétique des cellules germinales mâles dans le testicule fœtal bovin

Author

André, Marjolaine, Mandon-Pepin, Béatrice, Allais-Bonnet , Aurélie, Jammes, Hélène, Bonnet-Garnier, Amélie, Gelin, Valerie, Sellem, Eli, Pailhoux, Eric, Kiefer, Hélène, Pannetier, Maëlle, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Allice, and APISGENE programme GERMIMâLE

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

National audience

Published
2021

18. Fetal Estrogens are not Involved in Sex Determination But Critical for Early Ovarian Differentiation in Rabbits

Author

Jolivet, Geneviève, primary, Daniel-Carlier, Nathalie, additional, Harscoët, Erwana, additional, Airaud, Eloïse, additional, Dewaele, Aurélie, additional, Pierson, Cloé, additional, Giton, Frank, additional, Boulanger, Laurent, additional, Daniel, Nathalie, additional, Mandon-Pépin, Béatrice, additional, Pannetier, Maëlle, additional, and Pailhoux, Eric, additional

Published
2021
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20. Différenciation et reprogrammation épigénétique de la lignée germinale – Analyses transcriptomiques à l’échelle de la cellule

Author

Jolivet, Geneviève, Eloïse, Hérault, Allais-Bonnet, Aurélie, André, Marjolaine, Mandon-Pepin, Béatrice, Jouneau, Luc, Pailhoux, Eric, Pannetier, Maëlle, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Allice, and Pannetier, Maëlle

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019

22. L’épigénétique et la construction du phénotype chez le bovin

Author

Beaujean, Nathalie, Boutinaud, Marion, Devinoy, Eve, Jammes, Hélène, LE GUILLOU, Sandrine, Le Provost, Fabienne, Leroux, Christine, Mobuchon, Lenha, Pannetier, Maëlle, Sellem, Eli, Kiefer, Hélène, Biologie de la Reproduction, Environnement, Epigénétique & Développement (BREED), École nationale vétérinaire d'Alfort (ENVA)-Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut cellule souche et cerveau (U846 Inserm - UCBL1), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Physiologie, Environnement et Génétique pour l'Animal et les Systèmes d'Elevage [Rennes] (PEGASE), 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)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Génétique Animale et Biologie Intégrative (GABI), Université Paris-Saclay-AgroParisTech-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité Mixte de Recherche sur les Herbivores - UMR 1213 (UMRH), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Allice

Subjects
[SDV]Life Sciences [q-bio], [SDV.BA]Life Sciences [q-bio]/Animal biology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology
Abstract

International audience; In cattle breeding and farming, epigenetic mechanisms are regarded as a potential source of phenotypic variance unaccounted by genetic selection and could be used to better control the environmental impact on the genetic potential of animals. In this review, we briefly describe epigenetic processes, and address, from research studies mainly conducted at INRAE, the major role of epigenetic factors in the elaboration of phenotype in cattle. Focus is given to environmental factors shaping the epigenome, leading to variable effects on fertility, development, health and dairy production. The potential for using epigenetic markers in precision farming and as a complementary approach to genomic selection is also discussed.; En élevage bovin, une part importante de la variance phénotypique échappe à la sélection génétique, car l’expression des gènes quicontrôlent les caractères d’intérêt est déterminée par des facteurs à la fois génétiques et environnementaux. Une meilleure maîtrise desmécanismes épigénétiques, qui traduisent l’impact environnemental au niveau du génome, permettrait ainsi de mieux contrôler le phénotypedes animaux. Dans cette revue, nous décrivons brièvement les processus épigénétiques, puis nous évoquons, à travers plusieursexemples principalement issus de travaux menés à INRAE, le rôle majeur de l’épigénome dans la construction du phénotype chez le bovin.Nous mentionnons en particulier la façon dont certains facteurs environnementaux modifient les marques épigénétiques, ce qui entraînedes effets variables sur les caractères de fertilité, de développement, de santé et de production laitière. Le potentiel d’utilisation de marqueursépigénétiques dans le cadre d’un élevage de précision et en complément des analyses d’association entre génotype et phénotypemenées en sélection génomique est également discuté.

Published
2020

24. FOXL2 is a Progesterone Target Gene in the Endometrium of Ruminants

Author

Eozenou, Caroline, primary, Lesage-Padilla, Audrey, additional, Mauffré, Vincent, additional, Healey, Gareth D., additional, Camous, Sylvaine, additional, Bolifraud, Philippe, additional, Giraud-Delville, Corinne, additional, Vaiman, Daniel, additional, Shimizu, Takashi, additional, Miyamoto, Akio, additional, Sheldon, Iain Martin, additional, Constant, Fabienne, additional, Pannetier, Maëlle, additional, and Sandra, Olivier, additional

Published
2020
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25. Fetal Estrogens are not Involved in Sex Determination But Critical for Early Ovarian Differentiation in Rabbits.

Author

Jolivet, Geneviève, Daniel-Carlier, Nathalie, Harscoët, Erwana, Airaud, Eloïse, Dewaele, Aurélie, Pierson, Cloé, Giton, Frank, Boulanger, Laurent, Daniel, Nathalie, Mandon-Pépin, Béatrice, Pannetier, Maëlle, and Pailhoux, Eric

Abstract

AROMATASE is encoded by the CYP19A1 gene and is the cytochrome enzyme responsible for estrogen synthesis in vertebrates. In most mammals, a peak of CYP19A1 gene expression occurs in the fetal XX gonad when sexual differentiation is initiated. To elucidate the role of this peak, we produced 3 lines of TALEN genetically edited CYP19A1 knockout (KO) rabbits that were devoid of any estradiol production. All the KO XX rabbits developed as females with aberrantly small ovaries in adulthood, an almost empty reserve of primordial follicles, and very few large antrum follicles. Ovulation never occurred. Our histological, immunohistological, and transcriptomic analyses showed that the estradiol surge in the XX fetal rabbit gonad is not essential to its determination as an ovary, or for meiosis. However, it is mandatory for the high proliferation and differentiation of both somatic and germ cells, and consequently for establishment of the ovarian reserve. [ABSTRACT FROM AUTHOR]

Published
2022
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26. epigenome of male germ cells and the programming of phenotypes in cattle.

Author

Kiefer, Hélène, Sellem, Eli, Bonnet-Garnier, Amélie, Pannetier, Maëlle, Costes, Valentin, Schibler, Laurent, and Jammes, Hélène

Subjects
GERM cells, HISTONES, CHROMATIN, CATTLE fertility, PHYSIOLOGY, REPRODUCTIVE technology, PHENOTYPES, CATTLE
Abstract

The Epigenome of Bull Sperm and Its Relationships with Fertility Mature bovine spermatozoa have a particularly low global level of 5meC compared with bovine somatic cells and also to spermatozoa from goats, rams, humans, stallions, boars, and mice. Using a different whole-genome approach (whole-genome bisulfite sequencing), another team reported that the DNA methylome of bull sperm contains specific undermethylated domains enriched for satellites and evolutionary young TEs that may escape piRNA-mediated silencing ([62]). Comparative whole genome DNA methylation profiling of cattle sperm and somatic tissues reveal s striking hypomethylated patterns in sperm. The somatic epigenome represents the main barrier to the efficiency of cloning, while the oocyte and sperm epigenomes are extensively reprogrammed after fertilization to allow development of the embryo (Figure 3A). [Extracted from the article]

Published
2021
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27. Sdy, the unusual rainbow trout sex determination gene, hijacked the canonical vertebrate gonadal differentiation pathway

Author

Bertho, Sylvain, Herpin, Amaury, Branthonne, Adèle, Jouanno, Elodie, Yano, Ayaka, Nicol, Barbara, Müller, Thomas, Pannetier, Maëlle, Pailhoux, Eric, Miwa, Miwako, Yoshizaki, Goro, Schartl, Manfred, Guiguen, Yann, 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 ), University of Würzburg = Universität Würzburg, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Tokyo University of Marine Science and Technology (TUMSAT), and BBSRC Roslin Institute. GBR.

Subjects
[SDV]Life Sciences [q-bio], [INFO]Computer Science [cs], ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2019

28. A novel evolutionary conserved mechanism of RNA stability regulates synexpression of primordial germ cell-specific genes prior to the sex-determination stage in medaka

Author

Herpin, Amaury, Schmidt, Cornelia, Kneitz, Susanne, Gobé, Clara, Regensburger, Martina, Le Cam, Aurélie, Montfort, Jérome, Adolfi, Mateus C., Lillesaar, Christina, Wilhelm, Dagmar, Kraeussling, Michael, Mourot, Brigitte, Porcon, Béatrice, Pannetier, Maëlle, Pailhoux, Eric, Ettwiller, Laurence, Dolle, Dirk, Guiguen, Yann, Schartl, Manfred, 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), 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 National de la Recherche Scientifique (CNRS), Department of Anatomy & Neuroscience [Melbourne], School of Biomedical Sciences [Melbourne], Faculty of Medicine, Dentistry and Health Sciences [Melbourne], University of Melbourne-University of Melbourne-Faculty of Medicine, Dentistry and Health Sciences [Melbourne], University of Melbourne-University of Melbourne, Department of Developmental Biology, Hagedorn Research Institute, University of Heidelberg, Comprehensive Cancer Center Mainfranken, Hagler Institute for Advanced Study and Department of Biology, Texas A&M University System, This work was supported by the Deutsche Forschungsgemeinschaft by a grant (SCHA 408/12-1, HE 7135/2-1) to AH and MS as well as Credits Incitatifs Phase 2015/Emergence to AH. AH was additionally funded by the project AquaCRISPR (ANR-16-COFA-0004-01), Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Heidelberg University, and ANR-16-COFA-0004,AquaCrispr,Optimization of the CRISPR/Cas9 knock-in technology and application in salmon and trout(2016)

Subjects
Male, Embryology, [SDV]Life Sciences [q-bio], RNA Stability, gène déterminant majeur du sexe, sex determination, Oryzias, Gene Expression, Biochemistry, gonade, histologie, Database and Informatics Methods, poisson, Animal Cells, Medicine and Health Sciences, Biology (General), 3' Untranslated Regions, détermination du sexe, Messenger RNA, Eukaryota, Gene Expression Regulation, Developmental, protéine liaison rna, Biological Evolution, oryzias latipes, Nucleic acids, Ovaries, RNA Recognition Motif Proteins, Osteichthyes, Vertebrates, Female, Cellular Types, Anatomy, Genital Anatomy, Sequence Analysis, expression des gènes, Research Article, Fish Proteins, QH301-705.5, Bioinformatics, Research and Analysis Methods, gonad, régulation de la transcription, reproduction, analyse de génome, Sequence Motif Analysis, Genetics, Animals, adrianichthyidae, RNA, Messenger, Gonads, medaka, Embryos, Reproductive System, Organisms, embryogénèse, Biology and Life Sciences, Cell Biology, Sex Determination Processes, Germ Cells, Fish, cellule germinale, gène dmrt1, RNA, germ-line cells, facteur de transcription, Developmental Biology, Transcription Factors
Abstract

Dmrt1 is a highly conserved transcription factor, which is critically involved in regulation of gonad development of vertebrates. In medaka, a duplicate of dmrt1—acting as master sex-determining gene—has a tightly timely and spatially controlled gonadal expression pattern. In addition to transcriptional regulation, a sequence motif in the 3′ UTR (D3U-box) mediates transcript stability of dmrt1 mRNAs from medaka and other vertebrates. We show here that in medaka, two RNA-binding proteins with antagonizing properties target this D3U-box, promoting either RNA stabilization in germ cells or degradation in the soma. The D3U-box is also conserved in other germ-cell transcripts, making them responsive to the same RNA binding proteins. The evolutionary conservation of the D3U-box motif within dmrt1 genes of metazoans—together with preserved expression patterns of the targeting RNA binding proteins in subsets of germ cells—suggest that this new mechanism for controlling RNA stability is not restricted to fishes but might also apply to other vertebrates., In medaka a duplicate of dmrt1 acts as master sex-determining gene, with tightly controlled expression pattern in the gonads. This study shows that two RNA-binding proteins with antagonistic properties target a sequence motif in the 3’-UTR (D3U-box), promoting either RNA stabilization in germ cells or degradation in the soma., Author summary The development of the gonads in vertebrates is mainly regulated by dmrt1, a master sex-determining gene that has a timely and spatially controlled gonadal expression pattern. In addition to transcriptional regulation, a sequence motif located in the 3′ UTR (D3U-box) mediates transcript stability of dmrt1 mRNAs. However, this regulation is complex, and the mechanisms are barely understood. Here, we analyse further dmrt1 regulation in the fish model medaka and show that two RNA-binding proteins with antagonizing properties target the D3U-box and promote either RNA stabilization in germ cells or degradation in the soma. The evolutionary conservation of the D3U-box motif within metazoan dmrt1 genes—together with preserved expression patterns of the targeting RNA binding proteins in subsets of germ cells—suggest that this new mechanism for controlling RNA stability is not restricted to fishes but might also apply to other vertebrates.

Published
2019

29. Reprogrammation épigénétique des cellules germinales mâles chez les ruminants

Author

Pannetier, Maëlle, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and programme APIS-GENE GERMIMâLE

Subjects
[SDV]Life Sciences [q-bio], [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, ComputingMilieux_MISCELLANEOUS, [SDV.BDD.GAM]Life Sciences [q-bio]/Development Biology/Gametogenesis
Abstract

National audience

Published
2018

30. DNA methylation dynamics during spermatogenesis in ruminants

Author

Allais-Bonnet , Aurélie, Messiaen, Sébastien, André, Marjolaine, Mandon-Pepin, Béatrice, Livera, Gabriel, Schibler, Laurent, Jammes, Hélène, Pailhoux, Eric, Kiefer, Hélène, Pannetier, Maëlle, ProdInra, Migration, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Université Paris Saclay (COmUE), Institut National de la Santé et de la Recherche Médicale (INSERM), Département R&D, Allice, Grant APIS-gENE 2017-2020, Gordon Research Conferences (GRC). USA., and École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology
Abstract

International audience; DNA methylation of cytosines is a critical epigenetic modification in mammals that plays crucial roles in transcriptional regulation, chromatin remodelling and genomic imprinting. Dynamic erasure and reestablishment of DNA methylation marks are required for spermatogenesis and the normal function of mature sperm. DNA methylation is catalysed by DNA methyltransferase enzymes (DNMT) providing either maintenance (DNMT1) or de novo (DNMT3A/B/L) DNA methylation processes. The DNA methylation dynamics during spermatogenesis has been previously described in mice and humans but nothing is known in productive livestock. However, these investigations in ruminants could be helpful to determine epigenetic events likely to be crucial for male fertility. Interestingly, a recent study from our laboratory pointed DNA undermethylation of bull spermatozoa compared to other mammals such as humans, mice, sheep or goats [1]. This result raises the question of the dynamic of DNA methylation in bovine male germline. We thus propose to define and compare the methylation dynamics of bovine and caprine germ cells during spermatogenesis. Analyses of 5mC immunohistochemistry were performed on adult testis sections to illustrate the presence of DNA methylation marks in various spermatogenic cells. As purification of the different spermatogenic populations is required to characterize DNA methylation more precisely, we firstly developed a flow cytometry-based method (Hoechtst-FACS) for cell sorting from goat testis. Fraction enrichments were evaluated using stage-specific markers by real time qPCR analyses. In addition, de novo DNA methyltransferase expression was studied on the different fractions of purified spermatogenic cells, giving some clues on DNA methylation dynamics. In the future, spermatogenic cells will be purified from both caprine and bovine testes, and together with gene expression analyses (RNA-sequencing), DNA methylation will be determined for each cell types by Reduced-Representation Bisulfite Sequencing (RRBS)

Published
2018

31. The bizarre master sex determinant of salmonids triggers its action by directly hijacking the conserved gonadal differentiation pathway

Author

Bertho, Sylvain, Herpin, Amaury, Dupin-De-Beyssat, Elodie, Yano, Ayaka, Muller, T., Nicol, Barbara, Pannetier, Maëlle, Pailhoux, Eric, Schartl, M., Guiguen, Yann, 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), Julius-von-Sachs-Institute - Department of Molecular Plant Physiology and Biophysics, 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), Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and Instituto Nacional de Pesquisas da Amazônia (INPA). Manaus, BRA.

Subjects
fish, salmonidae, différenciation du sexe, [SDV]Life Sciences [q-bio], education, gène déterminant majeur du sexe, physiologie de la reproduction, reproduction, poisson, salmonids, transcription factors, transcription, différenciation de gonades, facteur de transcription, health care economics and organizations, expression des gènes
Abstract

The bizarre master sex determinant of salmonids triggers its action by directly hijacking the conserved gonadal differentiation pathway. 11. International Symposium on Reproductive Physiology of Fish

Published
2018

34. Epigenetic reprogramming in the male germ line of ruminants

Author

Mandon-Pepin, Béatrice, Allais-Bonnet , Aurélie, André, Marjolaine, CALVEL, Pierre, Sellem, Eli, Le Danvic, Chrystelle, Prézelin, Audrey, Jammes, Hélène, Pailhoux, Eric, Kiefer, Hélène, Pannetier, Maëlle, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Université Paris Saclay (COmUE), Département R&D, Allice, Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Institut National de la Recherche Agronomique (INRA), APIS-GENE 2017-2019, and ATouT. Com. FRA.

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2017

35. Reprogrammation de la méthylation de l’ADN des cellules germinales au cours du développement testiculaire chez Capra Hircus

Author

Allais-Bonnet , Aurélie, Mandon-Pepin, Béatrice, André, Marjolaine, Pailhoux, Eric, Prézelin, Audrey, Kiefer, Hélène, Pannetier, Maëlle, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Département R&D, CRYOLOG S.A, Financement Apis-Gene EGER 2017-2019, Biologie du développement et reproduction (BDR), and École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
cellules germinales, cellule germinale, ruminants, méthylation de l'adn, goat, testicule, germ-line cells, reprogrammation epigénétique, caprin, [SDV.BDD]Life Sciences [q-bio]/Development Biology
Abstract

Chez les mammifères, lors de leur différenciation, les cellules germinales font l’objet d’une reprogrammation de leur épigénome. Un effacement quasi total de la méthylation de l’ADN est observé dans les cellules germinales, lorsque celles-ci colonisent les futures gonades. Certaines régions du génome conservent néanmoins de la méthylation à des degrés divers en fonction du sexe des cellules germinales (XX ou XY) et de l’espèce considérée [1,2]. Ces régions où la méthylation est « non-effacée » pourraient alors être porteuses d’informations transgénérationnelles. L’apposition d’une nouvelle signature épigénétique spécifique du sexe des cellules germinales se met ensuite en place à des stades différents dans le testicule (vie fœtale) ou l’ovaire (après la naissance) [3]. Ainsi, chez les mâles, la re-méthylation de l’ADN débute pendant la vie in utero et l’environnement maternel peut donc conditionner l’acquisition de l’information épigénétique qui sera transmise, plus tard, par les spermatozoïdes de l’adulte à sa descendance. Afin de mieux connaitre les étapes clés de la re-méthylation du génome chez les espèces d’élevage, nous nous proposons de caractériser la reprogrammation des cellules germinales mâles comparativement chez trois espèces de ruminants : bovin, ovin et caprin. Des analyses globales ont déjà permis de déterminer des stades d’intérêts, notamment dans l’espèce caprine. Afin de pouvoir analyser la méthylation de l’ADN à ces différents stades, une population de cellules germinales la plus pure possible doit être isolée à partir de testicules fœtaux. Différentes techniques ont été testées, notamment la microdissection par capture laser (LCM, Laser Capture Microdissection). Un second challenge consistera ensuite à miniaturiser les procédés d’analyse, notamment ceux concernant l’étude de la méthylation (LCM-RRBS). L’extraction simultanée de l’ADN et de l’ARNm issus des cellules germinales microdisséquées permettra de comparer l’état épigénétique et transcriptomique des cellules germinales XY au cours des différentes étapes de leur différenciation.

Published
2017

36. In vitro studies of the long-range regulation of FOXL2 expression by epigenome editing

Author

Gobe, Clara, Daniel-Carlier, Nathalie, Jolivet, Geneviève, André, Marjolaine, Pailhoux, Eric, Pannetier, Maëlle, 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), Université Paris-Saclay, Department of Animal Physiology of the INRA (CI PHASE 2016), Biologie du Développement et Reproduction (BDR), and École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)

Subjects
FOXL2, in vitro study, expression génique, epigenome editing, ovaire, gene expression, CRISPR/dCas9, ovary, enhancer, épigénome, [SDV.BDD]Life Sciences [q-bio]/Development Biology, étude in vitro
Abstract

It has been shown that Forkhead boxL2 gene (FOXL2) is a master gene for female gonadal development. Firstly, it is involved in ovarian determination[1] and/or maintenance[2], and secondly it is necessary for ovarian follicle growth[3]. Interestingly, long-range deletions affecting FOXL2 expression have been reported in different species. In goats, a natural mutation called PIS for “Polled Intersex Syndrome” is responsible for female-to-male sex-reversal in homozygous XX PIS-/- mutant animals. This mutation corresponds to a 11.7kb homozygous deletion[4] lying at around 300 kb upstream of FOXL2 gene[5,6]. The PIS mutation is responsible for FOXL2 expression silencing and the sex-reversal[1]. In humans, various heterozygous long-range deletions have been shown to be responsible for decreasing FOXL2 expression, resulting in Blepharophimosis Ptosis Epicanthus inversus Syndrome[7]. To date, the Smallest Region of deletion Overlap (SRO) described in human patients, is a 7.4 kb region, encompassing the long non-coding RNA PISRT1 that was shown, in the goat species, to be closely regulated with FOXL2. All this data suggest that this distant region located at around 300 kb of FOXL2 may contain crucial regulatory elements for its expression. Analyzes of the epigenetic state of this region in human ovary (ChIP-Seq, ENCODE data), or of its counterpart in mouse ovary (ChIP-qPCR, personal data) revealed H3K27ac enrichment, which is an epigenetic mark preferentially observed on active gene enhancer or promoter. In order to prove that this region contains crucial enhancer(s), but also to demonstrate that long-range epigenetic modifications are able to control FOXL2 expression, we developed a program of epigenome editing. The CRISPR/dCas9 technology has been previously deflected to induce epigenetic modifications at specific loci[8]. Thus we designed several guide RNAs, coupled to a CRISPR/dCas9-p300 which is capable to acetylate H3K27. In a cellular model, different regions will be targeted, focusing on the most conserved elements of the SRO counterpart.

Published
2017

37. In vitro studies of the distal regulation of FOXL2 expression by epigenome editing

Author

Gobe, Clara, Daniel-Carlier, Nathalie, André, Marjolaine, Thépot, Dominique, Jolivet, Geneviève, Pailhoux, Eric, Pannetier, Maëlle, 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), Université Paris-Saclay, INRA Phase, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and Centre National de la Recherche Scientifique (CNRS). FRA.

Subjects
long-range regulation, FOXL2, in vitro study, epigenome editing, ovaire, CRISPR/dCas9, ovary, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, étude in vitro
Abstract

Prix du meilleur poster aux journées ReproSciences 2017; [i]In vitro[/i] studies of the distal regulation of [i]FOXL2[/i] expression by epigenome editing. 2. Journées Scientifiques du GdR Repro

Published
2017

38. 'SdY', the bizarre master sex determinant of salmonids, triggers its action by directly hijacking the conserved gonadal differentiation pathway

Author

Bertho, Sylvain, Herpin, Amaury, Dupin-De-Beyssat, Elodie, Yano, Ayaka, Muller, Thomas, Nicol, Barbara, Pannetier, Maëlle, Pailhoux, Eric, Schartl, Manfred, Guiguen, Yann, 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), Unité de recherche Développement de l'Elevage (LRDE), Institut National de la Recherche Agronomique (INRA), 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), Physiological Chemistry, Biocenter, Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Comprehensive Cancer Center Mainfranken, University Clinic Würzburg, Texas Institute for Advanced Study and Department of Biology, Texas A&M University System, Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Laboratoire de Recherches sur le Développement de l'Elevage (LRDE), Biologie du Développement et Reproduction (BDR), and École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)

Subjects
fish, salmonidae, reproduction, détermination du sexe, poisson, salmonids, [SDV]Life Sciences [q-bio], gène déterminant majeur du sexe, sex determination, différenciation de gonades, expression des gènes
Abstract

"SdY", the bizarre master sex determinant of salmonids, triggers its action by directly hijacking the conserved gonadal differentiation pathway. 1. European Symposium on Sex Determination in Vertebrates

Published
2017

39. Long range regulation of FOXL2 expression by epigenome editing

Author

Gobe, Clara, Daniel-Carlier, Nathalie, André, Marjolaine, Thépot, Dominique, Jolivet, Geneviève, Pailhoux, Eric, Pannetier, Maëlle, 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), Université Paris-Saclay, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and Institut National de la Santé et de la Recherche Médicale (INSERM). FRA. Medical Research Council (MRC), GBR. Institut Pasteur [Paris], FRA. Université de Genève (Unige), CHE. Institut National de la Recherche Agronomique (INRA), FRA.

Subjects
education, gène foxl2, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, développement folliculaire, épigénome, [SDV.BDD]Life Sciences [q-bio]/Development Biology, reproductive and urinary physiology, health care economics and organizations, humanities
Abstract

Long range regulation of [i]FOXL2[/i] expression by epigenome editing. 1. European Symposium on Sex Determination in Vertebrates

Published
2017

40. Additional file 1: Fig. S1. of An Assessment of Fixed and Native Chromatin Preparation Methods to Study Histone Post-Translational Modifications at a Whole Genome Scale in Skeletal Muscle Tissue

Author

Sarah-Anne David, Piégu, Benoît, Hennequet-Antier, Christelle, Pannetier, Maëlle, Aguirre-Lavin, Tiphaine, Crochet, Sabine, Bordeau, Thierry, Couroussé, Nathalie, Brionne, Aurélien, Bigot, Yves, Collin, Anne, and Coustham, Vincent

Abstract

Setup of chromatin preparation conditions. Table S1. ChIP-qPCR primers. Table S2. Quantity of chromatin and DNA used throughout the protocol. Fig. S2. Quality assessment of the sequencing reads. Fig. S3. Boxplot representing the distribution of the 40 M peaks lengths. Fig. S4. X-ChIP-seq and N-ChIP-seq peaks analysis. Table S3. Broad peak detection analysis by MACS2 for both H3K27me3 X-ChIP-seq and N-ChIP-seq experiments. Fig. S5. Comparison of H3K27me3-enriched peak regions detected by MACS2 and between MACS2 and epic. Supplemental methods X-ChIP-seq protocol for hypothalamus tissue samples. Table S4. Sequencing results of hypothalamus X-ChIP-seq analyzed using epic. Fig. S6. Fragmentation assessment of chromatin before immunoprecipitation for hypothalamus samples and X-ChIP results at three control loci. (PDF 1487 kb)

Published
2017
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41. Involvement of FOXL2 and RSPO1 in Ovarian Determination, Development, and Maintenance in Mammals

Author

Pannetier, Maëlle, Chassot, Anne-Amandine, Chaboissier, Marie-Christine, Pailhoux, Eric, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Université Paris Saclay (COmUE), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), ANR-13- BSV2-0017-02 ARGONADS & ANR-11-LABX-0028-01, and Fondation ARC pour la Recherche sur le Cancer (PJA 20131200236)

Subjects
Male, Mammals, endocrine system, rspo1, Ovary, sex determination, Forkhead Transcription Factors, Sex Determination Processes, Germ Cells, foxl2, Testis, Animals, Humans, Female, Thrombospondins, [SDV.BDD]Life Sciences [q-bio]/Development Biology, beta Catenin
Abstract

International audience; In mammals, sex determination is a process through which the gonad is committed to differentiate into a testis or an ovary. This process relies on a delicate balance between genetic pathways that promote one fate and inhibit the other. Once the gonad is committed to the female pathway, ovarian differentiation begins and, depending on the species, is completed during gestation or shortly after birth. During this step, granulosa cell precursors, steroidogenic cells, and primordial germ cells start to express female-specific markers in a sex-dimorphic manner. The germ cells then arrest at prophase I of meiosis and, together with somatic cells, assemble into functional structures. This organization gives the ovary its definitive morphology and functionality during folliculogenesis. Until now, 2 main genetic cascades have been shown to be involved in female sex differentiation. The first is driven by FOXL2, a transcription factor that also plays a crucial role in folliculogenesis and ovarian fate maintenance in adults. The other operates through the WNT/CTNNB1 canonical pathway and is regulated primarily by R-spondin1. Here, we discuss the roles of FOXL2 and RSPO1/WNT/ CTNNB1 during ovarian development and homeostasis in different models, such as humans, goats, and rodents.

Published
2016

42. Difference in fibroblasts’ ability to be reprogrammed by the oocyte cytoplasm: study of differential situation in bovine

Author

Dubé, Delphine, Kiefer, Hélène, Vitorino Carvalho, Anais, Aguirre-Lavin, Tiphaine, Pannetier, Maëlle, Le Bourhis, Daniel, Ruffini, Sylvie, Archilla, Catherine, Brochard, Vincent, Laffont, Ludivine, Gall, Laurence, Jouneau, Luc, Jammes, Hélène, Duranthon, Véronique, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Université Paris Saclay (COmUE), Station de Phénotypage, Allice, and International Organisation on Animal Reproduction. FRA.

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2016

43. Effects of low dose of bisphenol A on sheep fetal testis and ovary

Author

Panneau, Barbara, Mandon-Pepin, Béatrice, Pannetier, Maëlle, Jouneau, Luc, Cotinot, Corinne, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Agence Nationale de la Recherche (ANR) - Contrat MECABPA, and International Organisation on Animal Reproduction. FRA.

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2016

44. Reprogrammation épigénétique des cellules germinales mâles de ruminants : résultats et perspectives

Author

Perrier, Jean-Philippe, Sellem, Eli, Prézelin, Audrey, gasselin, Maxime, Jouneau, Luc, Piumi, Francois, Al Adhami, Hala, Weber, M., Renard, Jean Paul, Schibler, Laurent, Jammes, Hélène, Pannetier, Maëlle, Kiefer, Hélène, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Département R&D, Allice, Centre National de la Recherche Scientifique (CNRS), and Institut National de Recherche Agronomique (INRA). UAR Département Physiologie Animale et Systèmes d'Elevage (0558).

Subjects
[SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2016

45. An Assessment of Fixed and Native Chromatin Preparation Methods to Study Histone Post-Translational Modifications at a Whole Genome Scale in Skeletal Muscle Tissue

Author

David, Sarah-Anne, primary, Piégu, Benoît, additional, Hennequet-Antier, Christelle, additional, Pannetier, Maëlle, additional, Aguirre-Lavin, Tiphaine, additional, Crochet, Sabine, additional, Bordeau, Thierry, additional, Couroussé, Nathalie, additional, Brionne, Aurélien, additional, Bigot, Yves, additional, Collin, Anne, additional, and Coustham, Vincent, additional

Published
2017
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46. Regulator and target genes of FOXL2 in bovine endometrium

Author

Lesage Padilla, Audrey, Reinaud, Pierrette, Raliou, Mariam, Galio, Laurent, Bolifraud, Philippe, Giraud-Delville, Corinne, Al-Gubory, Kais Hussain, Charpigny, Gilles, Eozenou, Caroline, Pannetier, Maëlle, Sandra, Olivier, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), INRA-Phase Department, ED149 BIOSIGNE, and Ecole Doctorale Biosigne. FRA.

Subjects
foxl2, bovine, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, endometrium, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS
Abstract

National audience

Published
2015

47. Genetic cascade of sex determination in goats

Author

Pannetier, Maëlle, Boulanger, Laurent, Jouneau, Luc, Thépot, Dominique, Pailhoux, Eric, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), and The Endocrine Society. USA.

Subjects
mice, gonad differentiation, foxl2, sry, goat, sex determination, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, ComputingMilieux_MISCELLANEOUS, dmrt1
Abstract

International audience

Published
2015

48. [i]Dmxl2[/i] is involved in follicle formation in the mouse

Author

El Zaiat, Maëva, Gobe, Clara, Passet, Bruno, Meunier, Nicolas, André, Marjolaine, Jouneau, Luc, Congar, Patrice, Bonnet, Aurelie, Pailhoux, Eric, Pannetier, Maëlle, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Génétique Animale et Biologie Intégrative (GABI), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Neurobiologie de l'olfaction (NBO), Institut National de la Recherche Agronomique (INRA), ANR, FRM, Université de Haute Bretagne ( Rennes 2 ) (UR 2). Rennes, FRA., Neurobiologie de l'Olfaction et de la Prise Alimentaire (NOPA), and Institut National de la Recherche Agronomique (INRA)-AgroParisTech

Subjects
fertilité, endocrine system, folliculogénèse, allogreffes, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, létalité néonatale, [SDV.BDD]Life Sciences [q-bio]/Development Biology, dmxl2
Abstract

We identified DMXL2 from RNA-sequencing data as a gene preferentially expressed in differentiating-ovaries in the goat species. In order to determine its role during ovarian morphogenesis, we investigated functional studies in mice. Dmxl2 invalidation (KO) is lethal within the few hours following birth. The analysis of its expression in several tissues let us show that the heart, olfactory mucosa and brain expressed Dmxl2 during development. Our results further suggest that the lethality may be due to an alteration of the synaptic transmission.Besides, both male and female gonads express Dmxl2. Notably in the ovary, its expression increases few days before follicle formation. We show that DMXL2 protein is dynamically expressed in germ cells and granulosa cells; DMXL2 is strongly detected in the oocytes whereas its expression in the granulosa cells fluctuates depending on the stage of folliculogenesis. To study Dmxl2 implication during follicle formation, and to bypass the issue of lethality at birth, we performed allograft of KO ovaries on nude recipient mice. KO grafts show an alteration in follicle formation, followed by ovarian dysgenesis.To conclude, our work shows that Dmxl2 is involved in follicle formation in mice, and makes it a new candidate to explain some cases of premature ovarian failure. The conditional knock-out of Dmxl2 in germ cells or in granulosa cells is in progress in the laboratory (please refer to Clara Gobé’s poster) and will permit to precise its specific role in each cell types.

Published
2015

49. FOXL2 is a new progesterone-regulated gene in the endometrium

Author

Lesage Padilla, Audrey, Eozenou, Caroline, Healey, Gareth, Shimizu, Takashi, Oudin, Jean-Francois, Vaiman, Daniel, Myamoto, Akio, Sheldon, Martin, Reinaud, Pierrette, Charpigny, Gilles, Pannetier, Maëlle, Sandra, Olivier, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Swansea University, Obihiro University of Agriculture and Veterinary Medicine, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), ANR-08-GENM-037, INRA PHASE division and MESR, Université de Haute Bretagne ( Rennes 2 ) (UR 2). Rennes, FRA., and ProdInra, Migration

Subjects
[SDV.BDD] Life Sciences [q-bio]/Development Biology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology
Abstract

National audience; In mammals, mutual actions of estrogens and progesterone on their uterine receptors are essential for endometrium receptivity and conceptus implantation. In cattle we showed that FOXL2 -a key gene for ovarian differentiation and maintenance- is expressed and regulated in endometrium during oestrous cycle, a finding confirmed in murine and human endometrium. The present study aims to determine if FOXL2 is a progesterone-target gene in the bovine endometrium. Using various experimenta models in cattle, our results indicated (i) a negative correlation between FOXL2 gene expression and progesterone (P4) blood levels (ii) a significant reduction of FOXL2 transcript level in ovariectomized cows supplemented with P4 for 6 days (2.2-fold vs. control ovariectomized cows, P < 0.05) (iii) a significant decrease in FOXL2 mRNA level in bovine endometrial explants incubated with P4 (10-5 M) for 48h (2.4-fold vs. control explants, P < 0.05). No impact of oestradiol on FOXL2 gene expression was detected in these conditions. In order to confirm the regulation of FOXL2 promoter by P4, COS7 cells were transfected with a caprine FOXL2 reporter gene and progesterone receptor (PR) A or B expression vectors. In the presence of PRA and PRB, P4 (10-7 M) stimulated the activity of FOXL2 promoter (2.8-fold). Mutation of the P4 Response Element (PRE) in the caprine FOXL2 promoter abrogated the activity of this promoter in P4-treated COS7 cells overexpressing PRA/PRB. Collectively, our data show that reduced FOXL2 expression in the endometrium during the luteal phase results from the down-regulation of PRA/B known to occur in the presence of P4. Determining the biological actions of FOXL2 will be mandatory to define the contribution of this transcription factor in the regulation of sensor and driver properties of the endometrium.

Published
2015

50. Invalidation de Dmxl2 dans les cellules germinales femelles et diminution de la fertilité

Author

Gobe, Clara, El Zaiat, Maeva, André, Marjolaine, Allais-Bonnet , Aurélie, Pailhoux, Eric, Pannetier, Maëlle, Biologie du Développement et Reproduction (BDR), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), FRM, and Université de Haute Bretagne ( Rennes 2 ) (UR 2). Rennes, FRA.

Subjects
fertilité, cellule germinale, ovaire, invalidation conditionnelle, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, dmxl2
Abstract

National audience; Recent researches in our laboratory showed the implication of Dmxl2 gene in follicle formation in mouse ovaries (refer to Maëva Elzaiat’s poster). In the differentiating ovary Dmxl2 is expressed both by germ cells and somatic cells. Then, when folliculogenesis starts, the protein is detected in oocyte cytoplasm at every folliclar stages; but also in granulosa cells, faintly in primordial or primary follicles and strongly in follicle of later stages. Dmxl2 encodes for a scaffold protein presenting several protein/protein interaction domains, which appear to be involved in cell trafficking. In the female gonad, this protein could play a specific role in each cell types together with being involved in oocyte and granulosa cells communication. To determine the role of Dmxl2 in germ cells and in supporting cells, conditional invalidation of this gene is realized thanks to 2 transgenic lines of mice: one expressing the Cre recombinase under the control of the Vasa gene promoter in germ cells (Vasa-Cre); the other under the control of the AMH receptor II promoter in female supporting cells (Amhr2-Cre). Up to now, only mice invalidated for Dmxl2 in germ cells have been obtained. Regarding to our first results, females (Vasa-Cre; Dmxl2flox/flox) seem to be able to reproduce. However, ongoing fertility tests tend to show a lower fertility, (decreasing with age). Histological analyses of these mice ovaries at 7 weeks showed important mass cells and a few antrum follicles. Analyses at older stages are pursued. To conclude, invalidation of Dmxl2 in the germ cell line has no effect on initial follicle formation, but seems to be involved in ovarian function maintenance at adult stages.

Published
2015

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