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  • Single-vesicle architecture of synaptobrevin2 in astrocytes [Elektronski vir]
    Singh, Priyanka ...
    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.
    Source: Nature communications [Elektronski vir]. - ISSN 2041-1723 (Vol. 5, 2014)
    Type of material - e-article
    Publish date - 2014
    Language - english
    COBISS.SI-ID - 31384793

    Link(s):

    http://www.nature.com/ncomms/2014/140507/ncomms4780/full/ncomms4780.html
    DOI

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