Descrizione del progetto
Uno sguardo più approfondito a come le cellule «parlano» tra loro
Le molecole di adesione cellulare (CAM, Cell Adhesion Molecules) svolgono un ruolo importante nella strutturazione dell’architettura sinaptica, attraverso interazioni sfaccettate con vari partner sinaptici, e nell’orchestrazione della comunicazione neuronale. Il progetto CellCellEM, finanziato dall’UE, farà luce sul meccanismo che governa le attività sinaptiche delle CAM. Nello specifico, il progetto combinerà la microscopia crioelettronica a singola particella, la tomografia e metodologie biochimiche per esaminare i meccanismi molecolari delle latrofiline, le CAM che mediano la comunicazione cellula-cellula a livello di sinapsi. Gli studi si proporranno di fornire le basi strutturali per il funzionamento delle CAM e caratterizzeranno completamente l’insieme delle interazioni di latrofiline con le proteine nel sistema nervoso centrale. I risultati del progetto arricchiranno ulteriormente le nostre conoscenze relative alla comunicazione cellula-cellula nel sistema nervoso centrale.
Obiettivo
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.
Campo scientifico
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-STG - Starting GrantIstituzione ospitante
7610001 Rehovot
Israele