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Role of Endocytic Proteins in Mechanotransduction and Impact on Autosomal Dominant Centronuclear Myopathy Agathe Franck

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Role of Endocytic Proteins in Mechanotransduction and Impact on Autosomal Dominant Centronuclear Myopathy Agathe Franck Role of endocytic proteins in mechanotransduction and impact on autosomal dominant centronuclear myopathy Agathe Franck To cite this version: Agathe Franck. Role of endocytic proteins in mechanotransduction and impact on autosomal dom- inant centronuclear myopathy. Cellular Biology. Sorbonne Université, 2018. English. NNT : 2018SORUS453. tel-02926061 HAL Id: tel-02926061 https://tel.archives-ouvertes.fr/tel-02926061 Submitted on 31 Aug 2020 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. Université Pierre et Marie Curie École Doctorale Complexité du Vivant (ED 515) Institut de Myologie Équipe Physiopathologie & thérapie de la myopathie centronucléaire autosomique dominante Rôle des protéines de l’endocytose dans la mécanotransduction et la physiopathologie de la myopathie centronucléaire autosomique dominante Role of endocytic proteins in mechanotransduction and impact on autosomal dominant centronuclear myopathy Par Agathe Franck Thèse de doctorat Devant un jury composé de : Pr. Onnik Agbulut Président du jury & examinateur Dr. Jocelyn Laporte Rapporteur Dr. Athanassia Sotiropoulos Rapporteur Pr. Frances Brodsky Examinateur Dr. Stéphane Vassilopoulos Directeur de thèse To Lola Page | 3 Agathe Franck – phD thesis 2018 “On endocytic proteins in muscle” The woods are lovely, dark and deep, But I have promises to keep, And miles to go before I sleep, And miles to go before I sleep. — Robert Frost, Stopping by Woods on a Snowy Evening (1922) Page | 4 Disclaimer This thesis is delivered with a pair of red/cyan glasses for viewing of anaglyphs. The following logo means that you can wear the glasses for 3D viewing of the images: Please be aware that an absence of glasses does not impair reading of this thesis. Page | 5 Agathe Franck – phD thesis 2018 “On endocytic proteins in muscle” Acknowledgements First of all, I would like to thank Stéphane Vassilopoulos, my thesis supervisor. As I said, I’m not very good at this. Thank you for giving me a chance all these years ago, after reading the letter of a small master 1 student that just “wanted to do microscopy”. Thank you for the pep talks, the laughs, the evenings with Jeanne at the electron microscope, open mouthed in front of her beautiful pictures, the late lunches at the Japanese place near Jussieu, the pride I felt when I showed you my pictures, thank you for believing me, for your passion about our work, for maneuvering my boat through these years that have not always been easy… in short, thank you for everything. I’m also very grateful to Marc Bitoun, our great chief and leader. Thank you for your help, for always being caring and patient. Thank you also for your advices and very (very) pertinent questions. Jeanne, thank you forever for giving me the beautiful pictures that I included in this thesis manuscript. Thank you for your kindness and always being there when I needed to talk (most of the people I know think you are my shrink). You’ve helped me stay afloat and I will forever be grateful. How could I not thank lab 103. This melting pot of characters and personalities. Anaïs Fongy, Éline Lemerle, Corine, Margot, Bernard and Camilla. Sometimes we work there, but we also laugh. A lot. I’d like to thank especially Anaïs, who has helped me believe in myself at a time when no one could achieve it. Éline, I wish you all the best for the future, you can always call me. Anytime. I will also thank the expat members of team 2, Delphine and Gilles, and the one who is even further away, Bruno Cadot. I’m especially grateful to Gilles for the precious help and guidance since his arrival, and I am truly sorry for all the unlucky PCRs my cells have caused. Valérie Allamand, thank you for your reassuring presence in the lab, during evenings, during the weekends, thank you for your smile and for listening, and also… thank you for the milk! The YAP/TAZ part of this project could not have been realized without the help and expertise of Catherine Coirault. Thank you for the Flexcell and the various talks about YAP/TAZ shuttling, without you we would have been lost. This work was completed thanks to the receptiveness of the imaging platforms. A big thank you to the Cytometry and Imaging Platform of Gustave Roussy Institute, particularly to Corinne Laplace-Builhé and Sophie Salomé-Desnoulez. Thank you also to the imaging platforms of the Institut de Biologie Paris-Seine, Michaël Trichet for his enormous help in making metal replicas of our samples possible, and Susanne Bolte, Jean-François Gilles and France Lam for their expertise in fluorescence microscopy. I am also grateful to the Pitié-Salpêtrière cell imaging platform staff who often asked me if I wanted a bed installed next to their microscope. This work also relied on the biopsies given by patients affected by centronuclear myopathy or desminopathies. We are grateful to Muscle Tissue Culture Collection MTCC at Klinikum der Universität München for providing the p.R350P primary cells, and to Anne and Kamel of the Myology Institute’s immortalization platform for the immortalization of these cells and CNM patients’ myoblasts. I thank the Morphological Unit, especially Norma Romero, Michel Fardeau, Emmanuelle Lacène, Mai Thao Viou and Guy Brochier, for their precious help regarding the characterization of desmin anomalies in patients affected by AD CNM. Thanks also to Laura Julien of the vectorology platform regarding the production of the AAV used in this study. Pr. Onnik Agbulut, thank you for your smile, and for the precious help to this study by giving us desmin -/- mice. Page | 6 I am grateful to the members of thesis committee who have followed me through these years: Stéphanie Miserey-Lenkei, Guillaume Montagnac and Denis Furling. Stéphanie, I will not forget your precious advice. Marie-Claude Potier, thank you for your support, I carry a bit of the expertise acquired in your lab everywhere I go. I am grateful to the committee attributing doctoral contracts for believing in my project. I cannot end without thanking all the people who have carried me to hell and back: my mom, Mallory, Lucile, Emmanuelle, Georges, Clément and Guilhem Salines, Ronan, Aless, Sophie, the Gründ team, my favorite cousin Émilie, Alice, Julie, Gertrude, Florent, Capucine, Romain, Ségolène… This will seem strange but I’d like to thank my cat. He’ll understand. I owe more than a « thank you » to a person named Lola Salines. Lola, this work is dedicated to you, and I would like to write, as an opening to this thesis manuscript, all my gratitude, and all my love. Page | 7 Agathe Franck – phD thesis 2018 “On endocytic proteins in muscle” Table of contents Table of figures ____________________________________________________________ 10 Table of movies ____________________________________________________________ 12 List of tables ______________________________________________________________ 12 Abstract __________________________________________________________________ 13 French abstract ____________________________________________________________ 14 Abbreviations _____________________________________________________________ 15 I. Introduction ___________________________________________________________ 17 1- The skeletal muscle _________________________________________________________ 17 a. Function _________________________________________________________________________ 17 b. Development and Myofibers _________________________________________________________ 17 i. A striated pattern of contractile units _______________________________________________ 18 ii. Excitation-contraction coupling ____________________________________________________ 20 iii. Costameres ____________________________________________________________________ 21 iv. Intermediate filaments ___________________________________________________________ 22 c. Mechanotransduction ______________________________________________________________ 26 i. Integrin-mediated mechanotransduction ____________________________________________ 26 ii. YAP/TAZ pathway _______________________________________________________________ 28 iii. MRTF-SRF ______________________________________________________________________ 31 2- Membranes and vesicular trafficking __________________________________________ 32 a. The Cell Theory ___________________________________________________________________ 32 b. Membrane trafficking ______________________________________________________________ 33 c. Clathrin-mediated membrane traffic __________________________________________________ 35 i. Clathrin _______________________________________________________________________ 35 ii. Adaptor proteins ________________________________________________________________ 38 iii. Dynamin _______________________________________________________________________ 39 iv. Sequential recruitment of CCV formation actors _______________________________________ 42 v. Role of actin filaments ___________________________________________________________ 43 d. Diversity of clathrin-coated structures _________________________________________________ 44 i. A very stable assembly
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