Homotypic endothelial nanotubes induced by wheat germ agglutinin and thrombin

Scientific Reports
Lucia PediciniLynn McKeown

Abstract

Endothelial barrier formation is maintained by intercellular communication through junctional proteins. The mechanisms involved in maintaining endothelial communication subsequent to barrier disruption remain unclear. It is known that low numbers of endothelial cells can be interconnected by homotypic actin-driven tunneling nanotubes (TNTs) which could be important for intercellular transfer of information in vascular physiology. Here we sought insight into the triggers for TNT formation. Wheat germ agglutinin, a C-type lectin and known label for TNTs, unexpectedly caused striking induction of TNTs. A succinylated derivative was by contrast inactive, suggesting mediation by a sialylated protein. Through siRNA-mediated knockdown we identified that this protein was likely to be CD31, an important sialylated membrane protein normally at endothelial cell junctions. We subsequently considered thrombin as a physiological inducer of endothelial TNTs because it reduces junctional contact. Thrombin reduced junctional contact, redistributed CD31 and induced TNTs, but its effect on TNTs was CD31-independent. Thrombin-induced TNTs nevertheless required PKCα, a known mediator of thrombin-dependent junctional remodelling, suggesting a necess...Continue Reading

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Citations

Oct 20, 2018·Frontiers in Cell and Developmental Biology·Emil LouClifford J Steer

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Methods Mentioned

BETA
density gradient centrifugation
fluorescence microscopy
transfection
electrophoresis

Software Mentioned

OriginPro
OriginLab
FlexStation
OpenLab
DeltaVision
Origin
ImageJ
SoftWorx

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