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Characterisation of Weibel-Palade body fusion by amperometry in endothelial cells reveals fusion pore dynamics and the effect of cholesterol on exocytosis

Cookson, EA; Conte, IL; Dempster, J; Hannah, MJ; Carter, T (2013) Characterisation of Weibel-Palade body fusion by amperometry in endothelial cells reveals fusion pore dynamics and the effect of cholesterol on exocytosis. JOURNAL OF CELL SCIENCE, 126 (23). 5490 - 5499 (10). ISSN 0021-9533 https://doi.org/10.1242/jcs.138438
SGUL Authors: Carter, Thomas David

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Abstract

Regulated secretion from endothelial cells is mediated by Weibel–Palade body (WPB) exocytosis. Plasma membrane cholesterol is implicated in regulating secretory granule exocytosis and fusion pore dynamics; however, its role in modulating WPB exocytosis is not clear. To address this we combined high-resolution electrochemical analysis of WPB fusion pore dynamics, by amperometry, with high-speed optical imaging of WPB exocytosis following cholesterol depletion or supplementation in human umbilical vein endothelial cells. We identified serotonin (5-HT) immunoreactivity in WPBs, and VMAT1 expression allowing detection of secreted 5-HT as discrete current spikes during exocytosis. A high proportion of spikes (∼75%) had pre-spike foot signals, indicating that WPB fusion proceeds via an initial narrow pore. Cholesterol depletion significantly reduced pre-spike foot signal duration and increased the rate of fusion pore expansion, whereas cholesterol supplementation had broadly the reverse effect. Cholesterol depletion slowed the onset of hormone-evoked WPB exocytosis, whereas its supplementation increased the rate of WPB exocytosis and hormone-evoked proregion secretion. Our results provide the first analysis of WPB fusion pore dynamics and highlight an important role for cholesterol in the regulation of WPB exocytosis.

Item Type: Article
Additional Information: © 2013. Published by The Company of Biologists Ltd This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
Keywords: Amperometry, Endothelial cell, Exocytosis, Serotonin, VMAT1, Weibel–Palade bodies, Science & Technology, Life Sciences & Biomedicine, Cell Biology, CELL BIOLOGY, Weibel-Palade bodies, Endothelial cell, VMAT1, Serotonin, Amperometry, Exocytosis, VON-WILLEBRAND-FACTOR, PANCREATIC BETA-CELLS, VESICLE FUSION, FUNCTIONAL-CHARACTERIZATION, MONOAMINE TRANSPORTER, CATECHOLAMINE RELEASE, SEROTONIN TRANSPORTER, REGULATED EXOCYTOSIS, MEMBRANE CHOLESTEROL, VONWILLEBRAND-FACTOR, Developmental Biology, 06 Biological Sciences, 11 Medical And Health Sciences
SGUL Research Institute / Research Centre: Academic Structure > Molecular and Clinical Sciences Research Institute (MCS)
Academic Structure > Molecular and Clinical Sciences Research Institute (MCS) > Cell Sciences (INCCCS)
Journal or Publication Title: JOURNAL OF CELL SCIENCE
ISSN: 0021-9533
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Dates:
DateEvent
1 December 2013Published
PubMed ID: 24127569
Web of Science ID: 24127569
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URI: https://openaccess.sgul.ac.uk/id/eprint/103828
Publisher's version: https://doi.org/10.1242/jcs.138438

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