Androgen Signaling in Sertoli Cells Lavinia Vija To cite this version: Lavinia Vija. Androgen Signaling in Sertoli Cells. Human health and pathology. Université Paris Sud - Paris XI, 2014. English. NNT : 2014PA11T031. tel-01079444 HAL Id: tel-01079444 https://tel.archives-ouvertes.fr/tel-01079444 Submitted on 2 Nov 2014 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. UNIVERSITE PARIS-SUD ÉCOLE DOCTORALE : Signalisation et Réseaux Intégratifs en Biologie Laboratoire Récepteurs Stéroïdiens, Physiopathologie Endocrinienne et Métabolique Reproduction et Développement THÈSE DE DOCTORAT Soutenue le 09/07/2014 par Lavinia Magdalena VIJA SIGNALISATION ANDROGÉNIQUE DANS LES CELLULES DE SERTOLI Directeur de thèse : Jacques YOUNG Professeur (Université Paris Sud) Composition du jury : Président du jury : Michael SCHUMACHER DR1 (Université Paris Sud) Rapporteurs : Serge LUMBROSO Professeur (Université Montpellier I) Mohamed BENAHMED DR1 (INSERM U1065, Université Nice)) Examinateurs : Nathalie CHABBERT-BUFFET Professeur (Université Pierre et Marie Curie) Gabriel LIVERA Professeur (Université Paris VII) Marc LOMBÈS DR1 (INSERM U693, Université Paris Sud) PARIS SUD UNIVERSITY École Doctorale : Signalisation et Réseaux Intégratifs en Biologie Steroid Receptors, Metabolic and Endocrine Physiopathology Laboratory Reproduction and Development PhD THESIS Public dissertation on the 09/07/2014 By Lavinia Magdalena VIJA ANDROGEN SIGNALING IN SERTOLI CELLS Thesis Coordinator : Jacques YOUNG Professor (Université Paris Sud) Thesis Commitee : President of the jury: Michael SCHUMACHER DR1 (Université Paris Sud) Referees: Serge LUMBROSO Professor (Université Montpellier I) Mohamed BENAHMED DR1 (INSERM U1065, Université Nice) Examinators: Nathalie CHABBERT-BUFFET Professor (Université Pierre et Marie Curie) Gabriel LIVERA Professor (Université Paris VII) Marc LOMBÈS DR1 (INSERM U693, Université Paris Sud) Motto: “I am not searching for the answers, I am just trying to understand the questions” Confucius ACKNOWLEDGEMENTS It was a long journey finally ending with the dissertation. I would like to gratefully remember those people who supported me during this important period of my life. Dear Professor Jacques YOUNG, allow me to respectfully thank you for your support. From the moment you proposed to coordinate this project, you continued to generously share your professional experience and to teach me how to develop a scientific reasoning and how to discover the amazing universe of translational research. Thank you for having shared with me not only your clinical experience but also the seriosity and careful art of performing a bibliographic search and especially forhaving taught me the wisdom and the witty spirit and many other secrets necessary for writing a good scientific paper. Dear Marc LOMBES, words are not enough to express my gratitude for having so kindly adopted me in your research unit and for all the support you offered me during the last four years. Your permanent disponibility, your suggestions, always correct and pertinent to all my scientific or personal problems, were always remarkable. Thank you for all your help. I sincerely wish that this was just the beginning of a long journey in the universe of scientific research in molecular endocrinology, and that we shall continue new projects and new French-Romanian collaborations. Dear Professor Serge LUMBROSO and Doctor Mohamed BENAHMED allow me to respectfully thank you for all your support and understanding and for the enthusiasm you accepted to review this work. Words are not enough to express my respect and my gratitude for you. I would also like to sincerely thank Professor CHABBERT-BUFFET for all your help, support and time you shared so kindly with me during these last years; it is a pleasure and an honor for me that you accepted to be my thesis examinator. I would also like to thank Professor Gabriel LIVERA, for having so kindly accepted to be one of the examinators. I would also like to sincerely thank to all the members of the INSERM Unit 693, from Seniors (Professor AnneGUIOCHON-MANTEL, Hugues LOOSFELT, Nadine BINART, Jérôme B, Séverine, Jérôme F, Larbi and Damien, with a special thanks forSay- who always came up with a solution and support, even in those moments when experiments were not always associated with remarkable results, and who always showed a sign of friendship for everyone) to my collegues. Dear Adela, Bruno, Julien, Emmanuelle, Catherine, Junaid, Nathalie, Audrey, Anne-Lise, Ségolène, Jérôme N, and all students and collegues, with which I shared great moments, week-ends and long days of work, sometimes ending up at midnight, I am really happy I met you. Friendship is the most beautiful homeland as a song used to say and you have completely proven that this was true.Thank you, Geri MEDURI for having shared with me the secrets of the immunohistochemistry, for the magic of those great Italian dishes you prepare with so much love and for the pleasure of listening and supporting me.And last but not least, I would like to thank to all my friends and collegues from the “C.I. Parhon” National Institute of Endocrinology, as well as to all my colleagues and Seniors who allowed me to discover and to get competences in Nuclear Medicine (Professor Philippe Chaumet- Riffaud, Professor Alain Prigent, Frederique Archambaud and all the doctors from Bicêtre, StLouis and Avicenne Hospitals), to my family who understood my choices,as well as to all my friends from my both countries. TABLE OF CONTENTS Foreword 1 Introduction 1. Testicular differentiation and development 5 Factors involved in male sex differentiation A. Undifferentiated state 5 B. Gonadal differentiation 8 C. Hormonal control of the masculinization process 15 C.1.Introduction 15 C.2.Fetal testis development C.3.Fetal testicular hormonal milieu responsible for 16 masculinization C.4.Endocrine regulation of the fetal testis major cell types a) Leydig cells and testosterone production in human fetal testis 17 b) Fetal Sertoli cells 19 2. Human testicular physiology: seminiferous tubules and spermatogenesis 2.1. Testicular development from birth to adulthood 21 2.1.1.Postnatal Sertoli cells 21 2.1.2.Postnatal Leydig cells 24 2.1.3.Leydig cells and steroidogenesis 25 2.1.4.Postnatal germ cells and spermatogenesis 26 2.2 Hormonal regulation of spermatogenesis 28 2.2.1.Roles of FSH in spermatogenesis 29 2.2.2.Roles of androgens/LH in spermatogenesis 30 2.2.3.Roles of estrogens in spermatogenesis 33 2.2.4.Roles of thyroid hormones in spermatogenesis 34 2.2.5.Roles of insulin in spermatogenesis 35 3. Nuclear receptors(NRs): general structure and classification 3.1.Nuclear receptors: classification 36 3.2.Nuclear receptors: general structure 39 4 Androgens and the Androgen receptor(AR) 4.1Androgen action 40 4.1.1.Testosterone:biosynthesis and metabolism 41 4.1.2.Dihydrotestosterone:biosynthesis and metabolism 43 4.1.3.5-α-reductases 43 4.2.The Androgen Receptor (AR) 44 4.2.1. AR genomic signaling and effects 44 4.2.1.1. AR functional domains 45 a) The NH2-terminal domain (NTD) b) The DNA-binding domain (DBD) c) The hinge region d) The ligand binding domain (LBD) e) AR domains interactions 4.2.1.2.Transcriptional regulation of AR regulated target genes in testis 50 4.2.1.3.Posttranscriptional and posttranslational modifications of AR 50 1. AR Phosphorylation 2.Dephosphorylation 3. Acetylation 4. Methylation 5. SUMOylation 6. Ubiquitination Crosstalk between posttranslational modifications of AR 4.2.2.The Androgen Receptor:non genomic signaling and effects 55 4.3. The Androgen Receptor and the molecular biology of androgen 55 insensitivity 4.3.1Androgen insensitivity syndrome in humans 56 4.3.2. Androgen insensitivity syndrome in spontaneous and generated 59 AR-knockout models 5 Androgen receptor coregulators 60 5.1.Nuclear receptor(NR) coregulators 5.1.1.History of coregulators 60 5.1.2.General factors involved in coregulators functionality 61 5.1.3.Coactivators 65 5.1.4.Corepressors 66 5.1.5.Mechanism of interaction between steroid receptors(SR) and 67 coregulators 5.1.6.Coregulators involvement in biological processes 69 5.1.7.Coregulators and human disease 70 5.2.Androgen receptors (AR) coregulators 5.2.1.Introduction 71 5.2.2.Classification of AR coregulators 71 5.2.3.Modulation of AR transcriptional activation by androgen 72 receptor coactivators 5.2.4. Modulation of transcriptional repression by androgen receptor 78 corepressors 5.2.5.Animal knockouts and the biological roles of AR coregulators 80 5.2.6. Androgen receptor coregulators in human physiology and 81 pathology 5.2.6.1.Androgen receptor coregulators in androgen insensitivity 81 syndromes (AIS) and male infertility 5.2.6.2.Androgen receptor coregulators in prostate cancer 85 5.2.6.3.Androgen receptor coregulators in other pathologies 87 5.2.6.4. Androgen receptor coregulators: future directions 87 89 Objectives 97 Results First part:Androgenic regulation in Sertoli cells 99 Paper 1.Androgen-dependent stabilization of Androgen Receptor in the novel murine Sertoli cell line, ST38c Lavinia Vija, Kahina Boukari, Hugues Loosfelt, Geri Meduri, Say Viengchareun, Nadine Binart, Marc Lombès and Jacques YoungMol Cell Endocrinol. 2014 Mar 25;384(1-2):32-42 1.Introduction 101 2.Objectives 103 3.Discussion 105 4.Limits, further perspectives and conclusions 111 4.1. AR induced-AMH repression: hypothesis, results and perspectives 111 4.1.1.Introduction 111 4.1.2.Objectives 112 4.1.3.Results 113 4.1.4. Discussion 113 5.Conclusion 117 Second part: Androgen receptor coregulators (SRC-2 and HBO1): expression and characterization during testicular development Paper2.