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Nat Commun. 2015 Sep 24;6:8392. doi: 10.1038/ncomms9392.

Diffusional spread and confinement of newly exocytosed synaptic vesicle proteins.

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Leibniz Institut für Molekulare Pharmakologie (FMP) &Freie Universität Berlin, Department of Molecular Pharmacology and Cell Biology, Robert-Roessle-Straße 10, 13125 Berlin, Germany.
Freie Universität Berlin, Institut für Chemie und Biochemie, Takustrasse 6, 14195 Berlin, Germany.
Charite Universitätsmedizin, NeuroCure Cluster of Excellence, Virchowweg 6, 10117 Berlin, Germany.


Neurotransmission relies on the calcium-triggered exocytic fusion of non-peptide neurotransmitter-containing small synaptic vesicles (SVs) with the presynaptic membrane at active zones (AZs) followed by compensatory endocytic retrieval of SV membranes. Here, we study the diffusional fate of newly exocytosed SV proteins in hippocampal neurons by high-resolution time-lapse imaging. Newly exocytosed SV proteins rapidly disperse within the first seconds post fusion until confined within the presynaptic bouton. Rapid diffusional spread and confinement is followed by slow reclustering of SV proteins at the periactive endocytic zone. Confinement within the presynaptic bouton is mediated in part by SV protein association with the clathrin-based endocytic machinery to limit diffusional spread of newly exocytosed SV proteins. These data suggest that diffusion, and axonal escape of newly exocytosed vesicle proteins, are counteracted by the clathrin-based endocytic machinery together with a presynaptic diffusion barrier.

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