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Cholesterol regulates multiple forms of vesicle endocytosis at a mammalian central synapse
Author(s) -
Yue HaiYuan,
Xu Jianhua
Publication year - 2015
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/jnc.13129
Subject(s) - endocytosis , exocytosis , bulk endocytosis , endocytic cycle , microbiology and biotechnology , filipin , biology , kiss and run fusion , synaptic vesicle , pinocytosis , vesicle , cholesterol , biochemistry , cell , membrane
Abstract Endocytosis in synapses sustains neurotransmission by recycling vesicle membrane and maintaining the homeostasis of synaptic membrane. A role of membrane cholesterol in synaptic endocytosis remains controversial because of conflicting observations, technical limitations in previous studies, and potential interference from non‐specific effects after cholesterol manipulation. Furthermore, it remains unclear whether cholesterol participates in distinct forms of endocytosis that function under different activity levels. In this study, applying the whole‐cell membrane capacitance measurement to monitor endocytosis in real time at the rat calyx of Held terminals, we found that disrupting cholesterol with dialysis of cholesterol oxidase or methyl‐β‐cyclodextrin impaired three different forms of endocytosis, including slow endocytosis, rapid endocytosis, and endocytosis of the retrievable membrane that exists at the surface before stimulation. The effects were observed when disruption of cholesterol was mild enough not to change Ca 2+ channel current or vesicle exocytosis, indicative of stringent cholesterol requirement in synaptic endocytosis. Extracting cholesterol with high concentrations of methyl‐β‐cyclodextrin reduced exocytosis, mainly by decreasing the readily releasable pool and the vesicle replenishment after readily releasable pool depletion. Our study suggests that cholesterol is an important, universal regulator in multiple forms of vesicle endocytosis at mammalian central synapses.By monitoring endocytosis with whole‐cell membrane capacitance measurement, this study demonstrated that acute disruption of cholesterol with cholesterol oxidase or methyl‐β‐cyclodextrin impaired slow clathrin‐mediated endocytosis, rapid endocytosis, and endocytosis of pre‐existing retrievable membrane at a large mammalian nerve terminal, the calyx of Held. This data suggest a universal cholesterol requirement in distinct pathways of synaptic vesicle endocytosis.