Serotonin & Developmental Axonal Refinement

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Serotonin & Developmental Axonal Refinement PhD thesis University of Pierre and Marie Curie Specialty Physiology & Physiopathology Presented by Marta KOLODZIEJCZAK To obtain DOCTEUR de l’UNIVERSITÉ PIERRE ET MARIE CURIE Title: SEROTONIN & DEVELOPMENTAL AXONAL REFINEMENT: MICROGLIA CONTRIBUTION? Date of defense: 20 Mars 2015 Examining committee members: Ann Lohof President Etienne Audinat Referee Alexandre Dayer Referee Sonia Garel Examiner Anne Roumier PhD advisor Luc Maroteaux PhD director I II Acknowledgements First of all, I would like to express thanks to the jury members: the referees Etienne Audinat and Alexandre Dayer, the examiner Sonia Garel and the president Ann Lohof. It is a great pleasure and honor to be able to discuss my work with you. I am also grateful to Luc Maroteaux for welcoming me into his team. Thank you for creating such a friendly and stimulating environment and for your door always being open. You showed me the bigger picture of my research subject which pushed my thinking further and kept me enthusiastic about my work. I would like to express my heartfelt gratitude to my amazing supervisor Anne Roumier. Thank you that after these four years of work under your supervision I want to do science more than ever! Thank you for guiding me in a way that I could still make my own choices and that made me feel autonomous. Thank you that despite your very busy schedule you were always able to find time to discuss my work ensuring that I never felt left on my own. Thank you for showing me how to be a good scientist. Your motivation and joy of doing science is contagious and motivational. With you I learned how important it is to clearly define a scientific goal and persistently pursue it until getting some clear answers. You have amazing pedagogic skills and every student who gets the chance to work with you should feel truly lucky! I am grateful to Catherine Béchade for her encouraging and constructive feedback while writing my thesis. Catherine, you are an amazing scientist. Your help with the bibliography was invaluable! I also really appreciate your straightforwardness. It has been a great pleasure to do a collaboration with Nicolas Gervasi. Thank you for showing me the beauty of the live cells imaging. To see microglia moving around on a brain slice was one of the most spectacular experiments during my PhD! I acknowledge the invaluable help of Patricia Gaspar and members of her team: Alexanda Rebsam for help with the CTb experiments, Cedric Francius for the ISH and Ahlem Assali for her friendly advice as a more experienced PhD student. Thank you Aude, Sebastian, Maria, Mariano, Sophie & Teng Teng. It has been a great pleasure to have you all as lab neighbors. III I would like to express thanks to Michel Mallat and Christine Metin for their proficient advice during my mid-thesis committee. I would like to additionally thank Christine for providing me with the tomato reporter mouse which allowed me to generate some interesting results. I am indebted to: Mythili Savariradjane & Théano Eirinopoulou from the imaging platform. Mythili, thank you for the introduction to the amazing world of epifluorescence imaging (one of my favorite experimental tools). I really appreciated your always open door and the constant smile on your face. Théano, thank you for the extended lesson on confocal imaging and for the beautiful 3D reconstructions. I love your sense of humor. I would like to acknowledge the animal house stuff (especially Natacha) for their useful help despite difficult conditions. A big thank you to the former PhD student in our lab. Thank you Dr. Arno for introducing me to the graduate student life. I have always admire your joy and passion for science. The lab is not the same without you and your Bob Marley songs! I could not forget to mention the two amazing M2 students, Marine & Emily, with whom I quickly became friends. Marine, it was a great pleasure to have you around! Besides the protocols for DNA extractions you wrote; I also loved our little “pauses-cafés”. Ma chère Emily, I feel extremely lucky to have found a friend like you. My work days became much more enjoyable when you came around! The little favors you did for me over the past years (putting brains into sucrose, checking on my mice or sending me pictures of my lab notes) are countless… (I’ve lost the track of how many cakes I owe you! :P). I will miss our nights out and the “climbing Thursdays”! I would also like to thank other present and past members of the lab. Sophie, for her help with breeding animals. I have enjoyed our numerous discussions about scientific and non-scientific subjects. Stephane, for his help during the preparation of my PhD application. I am a big fan of the salad dressing you used to make at the lab “plateaux fromage” (never found out your secret…). Silvina, for the advice in animal care. It is amazing how one person can bring such an amount of positive energy to the whole office. Imane, for her very clear explanations and help with molecular biology. And also: Elodie, Damian & Takis. IV It has been an amazing journey and I will keep many warm memories from the years we worked together! Finally, but by no means least, I would like to thank my loved ones for their infinite support during all of these years. My friends: Chris, Emily, Maja, Ina, Ola & Milka for all the amazing trips, common meals & inspiring discussions. My family: Mum, Dad, Hubert, Grandma Renata, and all the others, thank you for always believing in me. Dear parents, thank you for letting me go my way even if at the time it sounded like a crazy idea. Your unconditional support gave me strength and courage to reach for the “impossible”. Mum, thank you for teaching me the courage to follow my dreams and not try to fit to some standards. Thanks to this precious knowledge I made sure I enjoyed every single moment of my PhD experience! I could not forget to thank my amazing boyfriend. Frederik, you have supported me all through the years of my PhD. Thanks to you, this journey turned out to be an amazing experience. Thank you for introducing me Matlab and for your huge help with image analysis. I love our life and our numerous amazing trips! I could not imagine a better life companion. I love you. V VI Table of Contents Acknowledgements ........................................................................................................................................ I Table of Contents .......................................................................................................................................... VI Table of Figures ............................................................................................................................................. XI List of acronyms ......................................................................................................................................... XIV Abstract .............................................................................................................................................................. 1 A - INDRODUCTION ....................................................................................................................................... 3 A1 Microglia .................................................................................................................................................. 5 A1.1 Microglia History ...................................................................................................................... 5 A1.2 Developmental Origin of Microglia ................................................................................... 6 A1.2.1 Microglia Originate from Extra-Embryonic Yolk Sac ............................................. 6 A1.2.2 Transition to the Committed Microglia Progenitors .............................................. 8 A1.2.3 Microglia Brain Colonization ........................................................................................... 9 A1.3 Microglia in Adult Brain ....................................................................................................... 11 A1.3.1 Origin of Microglia in Adult Brain ............................................................................... 11 A1.3.2 Adult Microglial Phenotype ........................................................................................... 13 A1.3.3 Microglia in Non-Pathological Conditions................................................................ 14 A1.3.4 Microglia Activation .......................................................................................................... 16 A1.3.5 Microglia Activation Markers ........................................................................................ 18 A1.3.6 Microglia vs. Macrophages ............................................................................................. 19 A1.4 Early Postnatal Microglia .................................................................................................... 21 A1.4.1 Developmental Microglial Phenotype ........................................................................ 21 A1.4.2 Developmental Neuronal Survival .............................................................................. 23 A1.4.3 Synaptogenesis, Synaptic Maturation & Plasticity during Development ..... 24 A1.4.4 Neurogenesis & Plasticity in Adult Brain ................................................................. 25 A2 Serotonin ..............................................................................................................................................
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