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Single-vesicle architecture of synaptobrevin2 in astrocytes.

Abstract：

:Exocytic transmitter release is regulated by the SNARE complex, which contains a vesicular protein, synaptobrevin2 (Sb2). However, Sb2 vesicular arrangement is unclear. Here we use super-resolution fluorescence microscopy to study the prevalence and distribution of endogenous and exogenous Sb2 in single vesicles of astrocytes, the most abundant glial cells in the brain. We tag Sb2 protein at C- and N termini with a pair of fluorophores, which allows us to determine the Sb2 length and geometry. To estimate total number of Sb2 proteins per vesicle and the quantity necessary for the formation of fusion pores, we treat cells with ATP to stimulate Ca2+-dependent exocytosis, increase intracellular alkalinity to enhance the fluorescence presentation of yellow-shifted pHluorin (YpH), appended to the vesicle lumen domain of Sb2, and perform photobleaching of YpH fluorophores. Fluorescence intensity analysis reveals that the total number of endogenous Sb2 units or molecules per vesicle is ≤25.

journal_name

Nat Commun

journal_title

Nature communications

authors

Singh P,Jorgačevski J,Kreft M,Grubišić V,Stout RF Jr,Potokar M,Parpura V,Zorec R

doi

10.1038/ncomms4780

subject

Has Abstract

pub_date

2014-05-07 00:00:00

pages

3780

issn

2041-1723

pii

ncomms4780

journal_volume

5

pub_type

杂志文章

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