Fiche Projet
Transcription
Fiche Projet
Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS Nom et prénom du directeur de thèse (et si besoin du) : Coudrier Evelyne, Le directeur de thèse et le co-directeur doivent impérativement être habilités à diriger les recherches (HDR) Coordonnées Tel : 0156246370 e-mail :[email protected] Nom et prénom du co-directeur : Del Bene Filippo Coordonnées Tel : 0156246552 e-mail : [email protected] Nom et prénom du responsable de l’équipe : Nombre de chercheurs et enseignants-chercheurs statutaires de l’équipe titulaires d’une HDR : 5 Nom et prénom du responsable du laboratoire : Bruno Goud Intitulé du laboratoire et N° d’unité : UMR 144 Spécialité : Biologie cellulaire-Biologie du développement Titre du projet de thèse : Myosin 1b for brain and axonal development Résumé du projet de thèse (1 page maximum, en anglais) Like all vertebrate Myosin 1 family members, Myosin 1b (Myo1b) has the capacity to couple mechanically the membrane with the actin cytoskeleton and generates mechanical forces. We have previously shown that Myo1b associated with organelles regulates membrane trafficking by controlling the morphology of these organelles and the formation of branched actin network (Almeida et al. 2011 Nature Cell Biol.). We also demonstrated that isolated Myo1b bound to a PI4,5P2 containing GUV (Giant Unilamellar Vesicle) can pull membrane tubes out of this GUV along actin bundles immobilized on a solid substrate (Yamada et al. 2014 Nature Com.). More recently we analyzed the role of Myo1b associated with the plasma membrane. We have demonstrated that Myo1b regulates the redistribution of myosin II in acto-myosin fibers and the formation of filopodia induced by EphB2 receptors stimulation (Prospéri et al. 2015 J. Cell Biol.). Together these data indicate that myosin1b controls various cellular processes including membrane trafficking and cell migration by linking mechanically membrane to the actin cytoskeleton and thereby controlling membrane shape. Our recent data indicate that Myo1b is also required for the formation of axons and the establishment of neuronal polarity in cultured hippocampal and cortical neurons by controlling the formation of actin waves. Furthermore, our biochemical data shows that Myo1b can physically interact with two proteins involved in axon guidance depending on netrin, UNC5 and MAX-1; MAX-1 being a member of a new protein family characterized by a PH and a FERM domains. The aim of our project is to reveal Myo1b function for the formation of axons and neuron polarity in relation with UNC5 and MAX-1 and elucidate the molecular mechanism by which it achieves its function in vivo. So far we have used as experimental model cultured neurons derived from post mitotic polarized neurons after dissociation from embryonic brain. Analysis of the polarization of this cellular model corresponds to the analysis of the repolarization of previously polarized neurons. In contrast most neurons undergo axon neurite polarization during migration in vivo. Thus, we propose to study the requirement of myo1b for brain and axonal development in vivo in zebrafish. This project will exploit the combination of cell biology approaches and genetic approaches that are currently developed in Evelyne Coudrier’s (Institut Curie UMR 144) and Filippo Del Bene’s (Institut Curie UMR 3215) teams. We have already generated a zebrafish loss-of-function model of Myo1b and we aim now to analyzed its phenotype in the initial axonal development in retina ganglion cells and in primary motoneurons, two cellular types for which we have the proper genetic tools to allow imaging studies and cellular dynamics in vivo. In particular we will analyze the consequence of Myo1b loss or overexpression of different mutated forms on initial axonal polarization, actin flow and endocytic compartment trafficking. Together these data will allow the establishment of Myo1b role in neuronal polarization and axonal extension. 1 Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS Thèses actuellement en cours dans l’équipe Nom et Prénom du doctorant Olga Iuliano Amal Kasri Guillaume Kulakowski Camilla Barros Charlotte Alibert Bruno Latche Nom du directeur de thèse Evelyne coudrier Bruno Goud -Stephanie Miserey (CR1) Bruno Goud -Stephanie Miserey (CR1) Kristin Schauer (HDR) Jean Batiste Menneville (HDR) Kristin Schauer Année de 1ere inscription et Ecole Doctorale 2011 2014 Financement pendant la thèse Institut Curie Paris VI 2014 ERC 2014 2014 sciencia sem fronteras ( financement du gouvernement brésilien) Paris VI programme IPV 2014 PSL Trois publications récentes du directeur de thèse (du co-directeur ou du co-encadrant s’il y a lieu).Mettre en gras le nom du directeur de thèse. Prosperi, M.T., Lepine P., Dingli F., Paul-Gilloteaux P., Martin R., Loew D., Knolker H.J., and Coudrier E. 2015. Myosin 1b functions as an effector of EphB signaling to control cell repulsion. J Cell Biol. 210:347-361. Yamada, A., Mamane A., Lee-Tin-Wah J., Di Cicco J., Prévost C., Lévy D., Joanny J.-F., Coudrier E., and Bassereau B. 2014. Catch bond behavior facilitates membrane tubulation by a non-processive myosin 1b Nature Communications. Apr 7;5:3624 Dunn TW, Gebhardt C, Naumann EA, Riegler C, Ahrens MB, Engert F, Del Bene F. 2016 Neural Circuits Underlying Visually Evoked Escapes in Larval Zebrafish. Neuron Feb 3;89(3):613-28. Docteurs encadrés par le directeur de thèse ayant soutenu après septembre 2010 et publications relatives à leur sujet de thèse. Mettre en gras le nom du directeur de thèse et celui du docteur. Nom Prénom : Lépine Priscilla 25/09/3013 Date de soutenance : Durée de thèse (en mois): 36 Ecole Doctorale : CDV-ED515 Publication : -Prosperi, M.T.*, Lepine P.*, Dingli F., Paul-Gilloteaux P., Martin R., Loew D., Knolker H.J., and Coudrier E. 2015. Myosin 1b 2 Ecole Doctorale COMPLEXITE DU VIVANT – Fiche Projet CONCOURS functions as an effector of EphB signaling to control cell repulsion. J Cell Biol. 210:347-361 -Lépine P., Revenu C. Prospéri MT, Del Bene F. Coudrier E. Myosin1b regulates the development of the intestinal epithelium in zebrafish (manuscrit soumis) Nom Prénom : Thomas Auer 30/09/2014 Date de soutenance : Durée de thèse (en mois): 36 Ecole Doctorale : Heidelberg University Germany (cotutelle Dr. Filippo Del Bene avec Prof. Jochen Wittbrodt) Publications : -Auer TO, Xiao T, Bercier V, Gebhardt C, Duroure K, Concordet JP, Wyart C, Suster M, Kawakami K, Wittbrodt J, Baier H, Del Bene F. Deletion of Kinesin I motor unmasks a mechanism of homeostatic branching control by Neurtrophin-3. eLIFE 2015 Jun 15;4 - Auer TO, Duroure K, Concordet JP, Del Bene F CRISPR/Cas9mediated conversion of eGFP into Gal4 transgenic lines in zebrafish. Nature Protocols 2014 Dec;9(12):2823-40. - Auer TO, Duroure K, De Cian A, Concordet JP, Del Bene F (2014) Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair. Genome research 24:142-153 3