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IN the PURSUIT of the FEAR ENGRAM: Identification of Neuronal Circuits Underlying the Treatment of Anxiety Disorder

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IN the PURSUIT of the FEAR ENGRAM: Identification of Neuronal Circuits Underlying the Treatment of Anxiety Disorder IN THE PURSUIT OF THE FEAR ENGRAM: Identification of neuronal circuits underlying the treatment of anxiety disorder THÈSE NO 7642 (2017) PRÉSENTÉE LE 2 NOVEMBRE 2017 À LA FACULTÉ DES SCIENCES DE LA VIE CHAIRE NESTLÉ - UNITÉ DU PROF. GRÄFF PROGRAMME DOCTORAL EN NEUROSCIENCES ÉCOLE POLYTECHNIQUE FÉDÉRALE DE LAUSANNE POUR L'OBTENTION DU GRADE DE DOCTEUR ÈS SCIENCES PAR Ossama Mohamed Salah El-Dien El-Sayed Ibrahim KHALAF acceptée sur proposition du jury: Prof. W. Gerstner, président du jury Prof. J. Gräff, directeur de thèse Prof. D. de Quervain, rapporteur Prof. S. Josselyn , rapporteuse Prof. C. Sandi, rapporteuse Suisse 2017 In the midst of his laughter and glee, He had softly and suddenly vanished away— For the Snark was a Boojum, you see. -Lewis Caroll The Hunting of the Snark (1876) by Lewis Carroll Cover of the first edition by Henry Holiday. U Acknowledgements Science may set limits to knowledge, but should not set limits to imagination. -Bertrand Russell Gibran Khalil Gibran once argued that a pundit spends his life in the pursuit of knowledge, but if one day he said I have had it all, then this would be the moment of his utmost ignorance. In life, in this very journey, we all share the same start of nascence and the inevitable destination of death, yet the path one takes to complete this voyage is essentially different from one another. Many factors could influence shaping these routes, and serendipity was central in moulding mine! It all started when I have first heard the story of Phineas Gage - the most famous brain-injury survivor - at the master’s defense of my eldest neurosurgeon brother. It was the first time for the regular pharmacist I used to be to start realizing that the brain is still a black box for us. It has lots of arcane details that we do not yet understand. This per se piqued my interest to the most and at that moment I decided to delve deeper seeking answers for the many questions started piling up. It was mainly the striking stories of Gage, patient H.M., and many others that incited me into quitting my job and dedicating my full time and concentration to search for answers to the many questions I had! And for that reason I would like to start appreciating them before those whom I have met in person during my journey and owe a big deal of recognition. During such pilgrimage, I am utterly grateful to many people who have helped me make headway in my expedition, and these people are: Prof. Johannes Gräff, my thesis advisor. I have enjoyed working under his tutelage and honoured to be the first member in his lab. Helping in constructing the lab from V scratch was an experience on the side that allowed me to see first hand how decent labs start and appreciate the effort all PIs put to start their own groups. His unconditional support and patience created the finest environment for me to thrive. His continuous guidance, and encouragement allowed me to develop my own ideas and skills. His knowledge and expertise were vital to overcome some bumps we had together in the road. His availability and openness for collaboration were some of the privileges I had and will always be indebted to. His dedication and enthusiasm is contagious and motivating, and made the period I spent in the lab more than enjoyable. He is setting the perfect example on how to be a successful yet modest scientist and a caring family man. The proper balance he achieved between managing his research group and his lovely small family is quite compelling, and I will always look up to him for that. Thanks for being the friend before being the supervisor. Profs. Wulfram Gerstner, Carmen Sandi, Dominique de Quervain, and Sheena Josselyn, thank you for accepting to be the jury for my PhD defense and have the time to read and evaluate my thesis and provide the proper feedback. I would also like to thank my candidacy exam committee: profs. Olaf Blanke, Bernard Schneider, and my great mentor prof. Harald Hirling, who was always available for me. The GräffLab In order of appearance, I would like to profoundly thank Mme. Liliane Glauser for her tremendous help in the lab. Her energy and dedication is fascinating. The great spirit of hers is indispensible. The great time and the laughs we had together were energizing. I would also like to immensely thank Mme. Soledad Andany for being the best lab assistant ever. Her remarkable support with all the paper work needed, and her shining smile were consequential part in my lab life. I have enjoyed our discussions, pauses, and strolls to the most. Dr. José Sanchez-Mut, besides being the big sib in the lab and his expertise were essential to propel my experiments, he is the brother I could have had in a parallel universe or another life if I were to be reborn Spanish there! His dedication, kindness, and cheerful personality are quite remarkable. I have learned a lot from him on the bench and even more in life outside of the lab - especially in the wilderness. He is the W kind of indispensable “Macgyver” friend one needs to have not only in the outdoor activities, but also the indoor ones! I have enjoyed doing lots of activities with him, during which we have had great discussions full of existential questions we tended to reflect upon together. For being a marvellous colleague, and a great friend, gracias caballero! Dr. Bianca Silva, the sister I never had, but lucky to have finally found and worked with. We have had lots of fun discussing science and doing other activities together. I feel grateful to have had this Italian bee whizzing near by. Her lively personality, sincere kindness, and cheerful smile were always refreshing during the toughest of times. I feel lucky also to have overlapped with the rest of the relatively new lab mates. I would like to thank all of them for the great support they were always providing especially Lucie Dixsaut for her tremendous help during her internship the first time she joined the lab, and her continuous support now upon re-joining the lab as a graduate student. She was central in many of the experiments I conducted during the doctorate years and I could not have had a better wingman to count on! Her unyielding feeling of hunger never prevented her from being there for me. Her jolly presence, and shrewd intellect were providing everything I could ask for. I would also like to thank my fellow graduate student Allie (aka Allison). Through her bubbly personality, radiant smile, and heartfelt spontaneity she proves everyday that she is the kind of vibrant spirit the lab needed long ago. I have enjoyed all of our discussions about worldly issues, as well as our regular bouldering schedule until I totaled my back! I would also like to thank the rest of my colleagues including Dr. Zimbul albo, and Fernanda Martínez Reza for providing just the right environment in the lab and for being convivial all the time. A final recognition for all the aforementioned lab members for the sacred schedule we used to have for our communal lunches. Respect for the team spirit! I have also had the pleasure to supervise and work closely with many students who have overlapped in our lab to conduct their master’s, or bachelor’s, or even a semester project. I would like to thank all of them for I have learned a lot from them and I have managed to hone my pedagogical methods because of their constructive feedbacks about the periods they have spent in the lab. I extend my particular appreciation to X Siegfried Resch, and Victoire Gorden for their ardent industry conducting their master’s project in close contact with me. They have proved to be very competent pushing forward their projects in the best possible way. I greatly appreciate working with them. I would also like to thank Ot Pratt, Ginna Urueña, Sébastien Hausmann, Florentine Barbey, and Margaux Silvestre for providing the helping hands when I needed them the most, and for being always motivated to conduct research in the lab. The EPFL core facilities The diligence and great attentiveness of many of the core facilities at the EFPL provided the best conditions for my project to progress. I am obliged for the help and support of the entire family of the center of PhenoGenomics (CPG) headed by Dr. Xavier Warot. I would also like to thank Dr. Emilie Gesina, Dr. Isabelle Desbaillets, Dr. Gisèle Ferrand, Dr. Raphaël Doenlen, and Dr. Roy Combe for their availability all the time and responsiveness concerning anything I needed for my animal work at the animal facility. I would also like to thank all the caretakers at the animal husbandries for paying great attention to the welfare of my animal lines, and keeping me updated all the time. The histology core facility (HCF) also played an important role in the progress of my project. I would, therefore, like to thank all the personnel working there especially the head of this unit Dr. Jessica Sordet-Dessimoz. Also the members of the Bioimaging and Optics Platform (BIOP) helped a lot fixing any problems I encountered either during the image acquisition process or even after during analysing them. I would like to thank for that Dr. Arne Seitz, Mr. José Artacho, Dr. Olivier Burri, Dr. Romain Guiet, Dr. Luigi Bozzo, and Mr. Thierry Laroche. For the cell-sorting experiments I would like to thank the Flow Cytometry Core Facility (FCCF) and its members Dr.
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