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The Scrib1 Interactome and Its Relevance for Synaptic Plasticity & Neurodevelopmental Disorders Vera Margarido Pinheiro

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The Scrib1 Interactome and Its Relevance for Synaptic Plasticity & Neurodevelopmental Disorders Vera Margarido Pinheiro The Scrib1 Interactome and its relevance for synaptic plasticity & neurodevelopmental disorders Vera Margarido Pinheiro To cite this version: Vera Margarido Pinheiro. The Scrib1 Interactome and its relevance for synaptic plasticity & neurode- velopmental disorders. Neurons and Cognition [q-bio.NC]. Université de Bordeaux, 2014. English. NNT : 2014BORD0318. tel-01435921 HAL Id: tel-01435921 https://tel.archives-ouvertes.fr/tel-01435921 Submitted on 16 Jan 2017 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. THÈSE PRÉSENTÉE POUR OBTENIR LE GRADE DE DOCTEUR DE L’UNIVERSITÉ DE BORDEAUX ÉCOLE DOCTORALE: SCIENCES DE LA VIE ET DE LA SANTE SPÉCIALITÉ: NEUROSCIENCES Par Vera PINHEIRO L’interactome de Scrib1 et son importance pour la plasticitè synaptique & les troubles de neurodéveloppement Sous la direction de : Nathalie SANS Soutenue le 4 Décembre 2014 Membres du jury : M. GARRET, Maurice CNRS, Bordeaux Président Mme CARVALHO, Ana Luisa Uni. Coimbra, Portugal rapporteur M. HENLEY, Jeremy Uni. Bristol, Royaume Uni rapporteur M. THOUMINE, Olivier CNRS, Bordeaux Examinateur M. BORG, Jean-Paul INSERM, Marseille Examinateur 2 Bordeaux University THESIS For the Doctorate of Bordeaux University Mention: Sciences, Technologies, Santé Option: Neurosciences Presented and defended publically on December 4th 2014 Vera PINHEIRO Born on 13th February 1987 at Portalegre, Portugal ______________________________________________________________________________________ 3 The Scrib1 Interactome and its relevance for synaptic plasticity & neurodevelopmental disorders ______________________________________________________________________________________ Jury members Dr. Maurice GARRET - Bordeaux, France- Chairman Prof. Ana Luisa CARVALHO - Coimbra, Portugal - Reviewer Prof. Jeremy HENLEY - Bristol, United Kingdom - Reviewer Dr. Olivier THOUMINE - Bordeaux, France - Examiner Prof. Jean Paul BORG - Marseille, France - Examiner Dr. Nathalie SANS - Bordeaux, France - Thesis supervisor Não há regresso. Há viagens sem regresso nem repetição. 4 Miguel Sousa Tavares, No teu deserto Acknowledgements First of all, I thank the jury members that accepted to evaluate the present work, honnouring me by their presence: Dr. Maurice Garret, Prof. Ana Luisa Carvalho, Prof. Jeremy Henley, Prof. Jean Paul Borg and Dr. Olivier Thoumine. I would like to thank SyMBAD (Marie Curie International Research and Training Program), Neurocentre Ma- gendie, Institut National de la Santé et de la Recherche Médicale, Conseil Regionel d’Aquitaine, Fondation pour la Recherche Médicale, Labex Brain and Association Schizo Oui for financial and/or logistical support. A special thank you to Pauline Lafenetre, Antonella Camitini, Poun Chea and Dania Prendin. A special thank you to Dr. Nathalie Sans and Dr. Mireille Montcouquiol for hosting me in the “Planar Cell Polarity and Plasticity”. I specially thank Dr. Nathalie Sans for your guide, patience and support throughout this last four years. Thank you as well to the past and present team members: Arnaud Giese, Elodie Richard, Christelle Durand, Miki Shimbo, Cédric Landmann, Aysegul Gezer, Tamrat Mamo, Lea Lasvaux, Maureen Decroo, Ronan Peyroutou, Benjamin Robert, Steve Carvalho dos Santos and Jerome Ezan. Lastly, an un- measurable thank you to Chantal Medina, Nicolas Piguel, Jean-Michel Blanc, Hortense Fanet, Muna Hilal- Larrieu and Maite Moreau, with whom I had the privilege to work aside and discuss with. Thank you as well to Amine Mehidi and Grégory Giannone, the Bordeaux Imaging Center, BioXtal and Giovanni Marsicano’s team, for sharing your knowledge and patience with me. 5 To all that I ran to across Europe before coming to Bordeaux (Universidade Nova de Lisboa, Universidade de Coimbra, Universittà degli Studi de Trieste). Antonio Laires, Palmira Costa, Cesar Fonseca, Carlos Du- arte, Enrico Tongiorgi and Gabriele Baj, thank you all for your guidance and helpful advices. To all those I had the chance to share a life-time experience with in the last four years: Philipp Bethge, Ber- nart Llinares i Gonzalez, Rihab Abdalhafiz, Ronan Chereau, Ciaran Murphy-Royal, Laurie Argaud, Ilaria Belluomo, Michele Colavita, Jimmy George, Laurent Ladepeche, Thomas Larrieu, Anael, Leslie & Julia Chazeau, Mikael Garcia, Lasani Wijetunge, Edgar Soria, Tiffany Desprez, José Cruz, Thomas Pfeiffer, Mar- tin Lenz, Caro & Serge. I hope to find you all somewhere, some day. And to those who, despite time and distance setting us apart, are always present: Frederike Lenz, Serra Yurur, Valentina Vaghi, Alessandro Fedele, Stefania Zulian, Giovanni Ferrati, Giacomo Benvenuti, Mehdi Bhouri, Elena Blanco, Boris, Vegas, Ana Quitéria, Guilherme Cardoso, Fabia Gomes, Joana Flores, Hélio Tomas, Claudia Correia, André Ferreria, Catarina Veiga, Liliana & Gonçalo Alexandre, Joao, Fernanda & José Lourenço, Felicia, Carlos & Lurdes Amador and last, but not the least, to my family. An exceptional thank you to an exceptional woman - my heroine, friend and mother, for your unconditional love and support. Finalement, à Désiré Mbida & Luca, merci de faire partie de ma nouvelle vie. Il ne me reste donc plus qu’un devoir, tourner la page et écrire un nouveau chapitre à votre côté. The Scrib1 Interactome and its relevance for synaptic plasticity & neurodevelopmental disorders The brain is made up of billions of nerve cells, or neurons, which can communicate with each other through functionally distinct structures - the axon and the dendrite. We focused our study on the development and maintenance of dendritic spines, whose changes in morphology are intimately correlated with synaptic plas- ticity, or the ability to respond to synaptic activity. Dendritic spines originate from motile dendritic filopodia, which mature into spines following axonal contact. However, the spatial and temporal coordination of all the molecular components throughout the formation and maturation of a synapse remains unclear. Scribble1 is planar cell polarity protein (PCP) classically implicated in the homeostasis of epithelial tissues and tumour growth. In the mammalian brain, Scrib1 is a critical scaffold protein in brain development and function. The main goal of this work was to investigate the molecular mechanisms underlying Scrib1 role in synapse formation and maintenance. Firstly, we depict the importance of Scrib1 PDZ-dependent interactions on glutamate receptors trafficking as well as bidirectional plasticity signalling pathway underying spatial memory. Secondly, we focus on the functional consequences of a recently identified autism spectrum disor- der (ASD) mutation of Scrib1 on neuronal morpholgy and function. We demonstrated that Scrib1 regulates dendritic arborization, spine formation and functional maintenance via an actin-dependent mechanism. Taken altogether, this thesis highlights the PCP protein Scrib1 as key scaffold protein in brain development and function, playing a plethora of roles from the subcelular to the cognitive level. 6 Keywords: Scrib1, autism spectrum disorders, hippocampal neurons, synaptic plasticity, dendritic spine, glutamate receptors traffic, actin dynamics. L’interactome de Scrib1 et son importance pour la plasticitè synaptique & les troubles de neurodéve- loppement Le cerveau contient environ cent milliards de neurones, qui communiquent entre eux par des structures fonc- tionnellement distinctes – l’axone et la dendrite Notre étude s’est intéressée au développement et au main- tien des épines dendritiques, dont les changements morphologiques sont intimement liés à la plasticité sy- naptique. Ces épines ont pour origine les filopodes qui évoluent en épines lors du contact axonal. Cette tran- sition implique une myriade de molécules, dont la coordination spatiale et temporelle reste largement mé- connue. Scribble1 est une protéine de polarité cellulaire classiquement impliquée dans l’homéostasie de tissues épi- théliaux, la croissance et progression des tumeurs. Scrib1 est aussi une protéine d’échafaudage critique pour le développement et le bon fonctionnement du cerveau. Notre objectif c’est été l’étude des mécanismes moléculaires sous-jacents à un rôle potentiel de Scrib1 dans la formation et le maintien des synapses. On a décrit l’importance d’interactions dépendantes des domaines PDZ sur le trafic des récepteurs glutamater- giques et sur la voie de signalisation de plasticité synaptique. Ensuite, nous avons évalué les conséquences fonctionnelles d’une mutation de Scrib1 récemment identifiée chez un patient humain atteint des troubles du spectre autistique dans la morphologie et fonction des neurones. L’ensemble de ce travail demontre que Scrib1, protéine d’échafaudage clé dans le développement et la fonction du cerveau, joue une multitude de rôle du niveau subcellulaire au niveau cognitif. 7 Mots clés: Scrib1, troubles du spectre autistique, neurones hippocampiques, plasticité synaptique, épine dendritique, trafic des récepteurs glutamatergiques, dynamique du cytosquelette d'actine. Physiopathologie de la plasticité neuronale Neurocentre Magendie INSERM U862 Université Bordeaux
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