Department of Anesthesiology

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Real-time measurements of vesicle-SNARE recycling in synapses of the central nervous system.

TitleReal-time measurements of vesicle-SNARE recycling in synapses of the central nervous system.
Publication TypeJournal Article
Year of Publication2000
AuthorsSankaranarayanan S, Ryan TA
JournalNat Cell Biol
Volume2
Issue4
Pagination197-204
Date Published2000 Apr
ISSN1465-7392
KeywordsAction Potentials, Animals, Cell Membrane, Cells, Cultured, Endocytosis, Fluorescent Dyes, Hippocampus, Hydrogen-Ion Concentration, Kinetics, Membrane Fusion, Membrane Proteins, Microscopy, Fluorescence, Nerve Tissue Proteins, Neurons, Presynaptic Terminals, R-SNARE Proteins, Rats, Rats, Sprague-Dawley, SNARE Proteins, Synaptic Vesicles, Vesicular Transport Proteins
Abstract

Following the fusion of synaptic vesicles with the presynaptic plasma membrane of nerve terminals by the process of exocytosis, synaptic-vesicle components are recycled to replenish the vesicle pool. Here we use a pH-sensitive green fluorescent protein to measure the residence time of VAMP, a vesicle-associated SNARE protein important for membrane fusion, on the surfaces of synaptic terminals of hippocampal neurons following exocytosis. The time course of VAMP retrieval depends linearly on the amount of VAMP that is added to the plasma membrane, with retrieval occurring between about 4 seconds and 90 seconds after exocytosis, and newly internalized vesicles are rapidly acidified. These data are well described by a model in which endocytosis appears to be saturable, but proceeds with an initial maximum velocity of about one vesicle per second. We also find that, following exocytosis, a portion of the newly inserted VAMP appears on the surface of the axon.

DOI10.1038/35008615
Alternate JournalNat. Cell Biol.
PubMed ID10783237
Grant ListNS24692 / NS / NINDS NIH HHS / United States