The crucial roles played by HNF1β during kidney development Filippo Maria Massa To cite this version: Filippo Maria Massa. The crucial roles played by HNF1β during kidney development. Human health and pathology. Université René Descartes - Paris V, 2012. English. NNT : 2012PA05T034. tel- 00771435 HAL Id: tel-00771435 https://tel.archives-ouvertes.fr/tel-00771435 Submitted on 8 Jan 2013 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. THESE DE DOCTORAT DE L’UNIVERSITE PARIS DESCARTES Ecole doctorale « Génétique, Cellules, Immunologie, Infectiologie, Développement », ED 157 Spécialité : Développement Présentée pour obtenir le titre de DOCTEUR de l’Université Paris Descartes “The crucial roles played by HNF1β during kidney development” Par M. Filippo Maria Massa Soutenance le 14 novembre 2012 Composition du jury: M. Marco Pontoglio Directeur de thèse Mme Evelyne Fischer Codirectrice de thèse M. Michel Cohen-Tannoudji Rapporteur Mme Brigitte Lelongt Rapportrice M. Bertrand Knebelmann Examinateur M. Gerard Waltz Examinateur M. Rémi Salomon Examinateur M. Weitzman Jonathan Examinateur Equipe "Expression Génique, Développement et Maladies" (EGDM) INSERM U1016/ CNRS UMR 8104 / Université Paris-Descartes Institut Cochin, Dpt. Génétique et Développement 24, Rue du Faubourg Saint Jacques, 75014 Paris, France « Considerate la vostra semenza: fatti non foste a viver come bruti ma per seguir virtute e canoscenza» Ulisse (Divina Commedia, Inferno canto XXVI, 118-120, Dante Alighieri) Aknowledgments - Remerciements – Ringraziamenti Four years of thesis are a long journey; fortunately I have be surrounded and supported by a lot of people that I will try to thank in this page, too short to be exhaustive for all the gratitude I have to them. In first place, I would like to thank the members of the jury that have kindly accepted to evaluate my work: Pr. Knebelmann, Pr Waltz, Pr Knebelman, Pr Salomon. A special thank goes to Mme Lelongt and M Cohen-Tannoudji to have reviewed my thesis and to give very useful suggestions in order to ameliorate it. A long journey needs good leading. For the precious and irreplaceable help and support I would like to thank Marco and Evelyne that overviewed all my work (and corrected the not- so-few mistakes I did on the way). They avoid me to get lost in failing experiments and frustrating periods, as well as to encourage my enthusiasm. I’m still in love with the research! Once the journey has started, I found a very important help from the entire group of colleagues: the “Pontoglio’s team” crew always supported me and offered me the pratical/technical/moral support to achieve my work. I would like to thank the actual members of the lab (Serge, Michel, Magali, Jon, Cecile and Claire) and the people I have the pleasure to meet in the lab and that are now in the different corner of the world (the spanish Miguel and Kiko, Celine and Tristan). A big thank to all the friends that I meet during my thesis: too many to number them without forgiving someone… Thanks for the support you gave me, for the antibodies protocols and suggestion you gave me, without forgetting the long nights spent partying around! In conclusion, useless to check the list of the names, I could not double the size of my thesis!! But I will remember all of you wherever I will go Of course, a very special thank goes to my parents Ezio and Grazia that supported me (even with excellent food supplies) all along my thesis and to my brothers Corrado and Emanuele! Thanks also to Giulia and Veronica to support and stand my bros, and without forgetting the newcomer Edoardo. Finally, THE very special thanks to Eva that stand me all along the way and gave me the strength to keep going on, even if sometimes the things seem not to work and I was tempted to give up. Thank youuuu!! F.Massa Hnf1b during kidney development Table of Contents Introduction I I. Kidney morphology and functions ................................................................................................... 3 Introduction II II. Principal signaling pathways during renal morphogenesis ........................................................ 12 The Notch Pathway ........................................................................................................................... 13 The Wnt signaling pathways ............................................................................................................. 17 Bone Morphogenic Proteins (BMPs) signaling pathways ................................................................. 22 Fibroblast Growth Factor (FGF) signaling pathway ......................................................................... 25 Introduction III III. Kidney development ..................................................................................................................... 28 1. Pronephros and mesonephros .................................................................................................... 30 Morphology ................................................................................................................................... 30 Molecular mechanisms .................................................................................................................. 31 2. Metanephros Development ......................................................................................................... 34 Metanephric mesenchyme identity. ............................................................................................... 34 The emergence of the ureteric bud. ............................................................................................... 40 Branching induction and regulation. ............................................................................................. 46 Metanephric mesenchyme condensation. ...................................................................................... 50 Mesenchyme derived structures formation.................................................................................... 51 Pre-Aggregates and Renal Vesicle formation ............................................................................... 51 Comma-shaped body development ............................................................................................... 56 S-shaped body development ....................................................................................................... 56 The mature nephron morphogenesis ............................................................................................... 57 1. Formation of the glomerulus ......................................................................................... 57 2. Tubular maturation ......................................................................................................... 59 3 F.Massa Hnf1b during kidney development Introduction IV IV. Transcription factors and gene expression ................................................................................. 71 1. Structure of transcription factors and their specificity ................................................................. 75 2. The Hepatocyte Nuclear Factor 1 family ...................................................................................... 77 3. The Hepatocyte Nuclear Factor 1 in the kidney ........................................................................... 82 Introduction V V. Hnf1 transcription factors and human diseases (MODY syndrome) ....................................... 86 1. Pancreatic defects correlated to MODY syndrome ....................................................................... 88 2. Renal defects correlated to MODY syndrome .............................................................................. 89 Project Design ............................................................................................................................... 93 Material and Methods .............................................................................................................. 96 Results Part I Role of Hnf1b in early step of kidney development and ureteric bud branching........................ 103 Results Part II Role of Hnf1b in tubulogenesis ......................................................................................................... 133 Discussion The role of Hnf1b role during kidney morphogenesis................................................................... 166 Perspectives Analysis of gene expression in Hnf1b deficient nephron precursors ............................................ 175 Role of Hnf1b during tubular specification and elongation.. ....................................................... 176 3D reconstruction of nephron precursors ..................................................................................... 178 Bibliography 4 Introduction I F.Massa Hnf1b during kidney development I. Kidney morphology and functions The kidney is a complex organ, located at the rear of the abdominal cavity in the retroperitoneum (figure 1A). The kidneys play crucial roles