Synaptotagmin 5 Regulates Ca-dependent Weibel-Palade Body Exocytosis in Human Endothelial Cells

Overview

Journal J Cell Sci

Specialty Cell Biology

Date 2019 Jan 20

PMID 30659119

Citations 9

Authors

Camille Lenzi

Jennifer Stevens

Daniel Osborn

Matthew J Hannah

Ruben Bierings

Tom Carter

Affiliations

Soon will be listed here.

Abstract

Elevations of intracellular free Ca concentration ([Ca]) are a potent trigger for Weibel-Palade body (WPB) exocytosis and secretion of von Willebrand factor (VWF) from endothelial cells; however, the identity of WPB-associated Ca-sensors involved in transducing acute increases in [Ca] into granule exocytosis remains unknown. Here, we show that synaptotagmin 5 (SYT5) is expressed in human umbilical vein endothelial cells (HUVECs) and is recruited to WPBs to regulate Ca-driven WPB exocytosis. Western blot analysis of HUVECs identified SYT5 protein, and exogenously expressed SYT5-mEGFP localised almost exclusively to WPBs. shRNA-mediated knockdown of endogenous SYT5 (shSYT5) reduced the rate and extent of histamine-evoked WPB exocytosis and reduced secretion of the WPB cargo VWF-propeptide (VWFpp). The shSYT5-mediated reduction in histamine-evoked WPB exocytosis was prevented by expression of shRNA-resistant SYT5-mCherry. Overexpression of SYT5-EGFP increased the rate and extent of histamine-evoked WPB exocytosis, and increased secretion of VWFpp. Expression of a Ca-binding defective SYT5 mutant (SYT5-Asp197Ser-EGFP) mimicked depletion of endogenous SYT5. We identify SYT5 as a WPB-associated Ca sensor regulating Ca-dependent secretion of stored mediators from vascular endothelial cells.

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