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Title:
STED microscopy reveals that synaptotagmin remains clustered after synaptic vesicle exocytosis
Authors:
Willig, Katrin I.; Rizzoli, Silvio O.; Westphal, Volker; Jahn, Reinhard; Hell, Stefan W.
Affiliation:
AA(Departments of NanoBiophotonics and), AB(Neurobiology, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany), AC(Departments of NanoBiophotonics and), AD(Neurobiology, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany), AE(Departments of NanoBiophotonics and)
Publication:
Nature, Volume 440, Issue 7086, pp. 935-939 (2006). (Nature Homepage)
Publication Date:
04/2006
Origin:
NATURE
Abstract Copyright:
(c) 2006: Nature
DOI:
10.1038/nature04592
Bibliographic Code:
2006Natur.440..935W

Abstract

Synaptic transmission is mediated by neurotransmitters that are stored in synaptic vesicles and released by exocytosis upon activation. The vesicle membrane is then retrieved by endocytosis, and synaptic vesicles are regenerated and re-filled with neurotransmitter. Although many aspects of vesicle recycling are understood, the fate of the vesicles after fusion is still unclear. Do their components diffuse on the plasma membrane, or do they remain together? This question has been difficult to answer because synaptic vesicles are too small (~40nm in diameter) and too densely packed to be resolved by available fluorescence microscopes. Here we use stimulated emission depletion (STED) to reduce the focal spot area by about an order of magnitude below the diffraction limit, thereby resolving individual vesicles in the synapse. We show that synaptotagminI, a protein resident in the vesicle membrane, remains clustered in isolated patches on the presynaptic membrane regardless of whether the nerve terminals are mildly active or intensely stimulated. This suggests that at least some vesicle constituents remain together during recycling. Our study also demonstrates that questions involving cellular structures with dimensions of a few tens of nanometres can be resolved with conventional far-field optics and visible light.
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