Structural and Molecular Decoding of Presynaptic Vesicle Biogenesis in situ
| Title | Structural and Molecular Decoding of Presynaptic Vesicle Biogenesis in situ |
|---|---|
| Acronym | PREBIOS |
| Start date | 2026-06-01 |
| End date | 2028-05-31 |
| Sponsor | Marie Skłodowska-Curie Action (MSCA) |
| Institution | Leibniz-Forschungsinstitut fuer Molekulare Pharmakologie im Forschungsverbund Berlin e.V. |
Associated cell lines
Project Description
Neurotransmission relies on the precise formation and maturation of synaptic vesicles (SVs), yet the early stages of their biogenesis remain poorly understood. Precursor vesicles (PVs) are the nascent carriers that transport SV and active zone proteins from the neuronal soma to presynaptic terminals, but their molecular composition, architecture, and formation mechanisms are unknown. Dysfunction in these processes is tightly linked to neurological disorders such as autism, epilepsy, and Parkinson's disease, highlighting both the fundamental and biomedical importance. This project aims to establish the first comprehensive structural and molecular framework of PVs. Building on the researcher's expertise in structural analysis of synaptic proteins and the host laboratory's pioneering work in presynapse formation, the action will pursue three objectives: (1) define the architecture and composition of isolated PVs from human iPSC/ESC-derived neurons using cryogenic electron microscopy and tomography (cryo-EM and cryo-ET), and quantitative proteomics; (2) visualize PVs in situ within axons through advanced correlative light and electron microscopy to uncover cargo identity and transport mechanisms; and (3) resolve the origin, remodelling, and maturation of PVs into SVs using pulse-chase labelling strategies and gene engineering verification. The methodology integrates structural biology, proteomics, and neuronal cell models to overcome the scarcity and transient nature of PVs. The anticipated outputs—structural atlases, proteomic datasets, and mechanistic models—will deliver unprecedented insight into presynaptic biogenesis. By resolving how PVs form, remodel, and mature into SVs, the project will close a major knowledge gap in neuroscience. The results will not only advance understanding of synaptic biology but also provide methodological innovations broadly applicable to cell biology and neuropharmacology, strengthening European leadership in neuro-structural research.