Description du projet
Observer de plus près la façon dont les cellules «se parlent»
Les molécules d’adhésion cellulaire (CAM) jouent un rôle important dans la structuration de l’architecture synaptique par le biais d’interactions à multiples facettes avec divers partenaires synaptiques et l’orchestration de la communication neuronale. Le projet CellCellEM, financé par l’UE, va faire la lumière sur le mécanisme régissant les activités synaptiques des CAM. Plus précisément, le projet combinera la cryo-microscopie électronique (cryo-ME) à particule unique, la tomographie et des méthodologies biochimiques pour disséquer les mécanismes moléculaires des latrophilines (LPHN) – les CAM qui assurent la communication entre cellules au niveau de la synapse. Les études viseront à fournir la base structurelle de la fonction des CAM et caractériseront entièrement l’ensemble des interactions des LPHN avec les protéines du système nerveux central (SNC). Les résultats du projet viendront enrichir nos connaissances sur la communication cellule-cellule au sein du SNC.
Objectif
Synapses are intercellular junctions specialized for coordinated cell-cell communication throughout the nervous system. They are organized by cell-adhesion molecules (CAMs) that bi-directionally orchestrate neuronal communication. Latrophilins (LPHNs) are a unique sub-family of CAMs that play critical roles in structuring the synaptic architecture through multifaceted interactions with a large variety of synaptic partners. Mutations in LPHN have been associated with neurodevelopmental and neuropsychiatric disorders. Despite their gravity, the mechanism governing LPHN synaptic activities remain elusive.
To further our understanding of LPHN-mediated cell-cell communication, we suggest to characterize these receptors’ interactions with their intracellular and extracellular partners. For this purpose, we propose to adopt a hybrid approach driven primarily by cryo-EM, a state-of-the-art technique capable of dissecting the molecular mechanisms of super-molecular assemblies at extremely high spatial resolutions, which is our group’s main field of expertise. The cryo-EM studies will be complemented by cryo electron tomography (cryo-ET), fluorescence microscopy and biochemical approaches. Our specific aims are:
Aim 1: Dissect the molecular mechanisms of LPHN activation by combining cryo-EM with biochemical methodologies.
Aim 2: Characterize the LPHN interactome through cryo-EM and fluorescence microscopy.
Aim 3: Resolve the architecture of the LPHN interactome at a close-to-native environment through cryo-ET.
Our experimental strategy will generate a quantitative, near-atomic resolution view of LPHNs and the mechanism by which they interact with their synaptic partners and instigate trans-synaptic signal transduction. These data will be vital for understanding LPHN-mediated cell-cell communication as well as the mechanisms governing trans-synaptic interactions and could potentially highlight novel approaches to treat neurodevelopmental and neuropsychiatric disorders.
Champ scientifique
Programme(s)
Thème(s)
Régime de financement
ERC-STG - Starting GrantInstitution d’accueil
7610001 Rehovot
Israël