Pressure-dependent regulation of Ca2+ signalling in the vascular endothelium

Wilson, Calum; Saunter, Christopher D.; Girkin, John M.; McCarron, John G.

Abstract

The endothelium is an interconnected network upon which haemodynamic mechanical forces act to control vascular tone and remodelling in disease. Ca2+ signalling is central to the endothelium's mechanotransduction and networked activity. However, challenges in imaging Ca2+ in large numbers of endothelial cells under conditions that preserve the intact physical configuration of pressurized arteries have limited progress in understanding how pressure-dependent mechanical forces alter networked Ca2+ signalling. We developed a miniature wide-field, gradient-index (GRIN) optical probe designed to fit inside an intact pressurized artery that permitted Ca2+ signals to be imaged with subcellular resolution in a large number (∼200) of naturally connected endothelial cells at various pressures. Chemical (acetylcholine) activation triggered spatiotemporally complex, propagating inositol trisphosphate (IP3)-mediated Ca2+ waves that originated in clusters of cells and progressed from there across the endothelium. Mechanical stimulation of the artery, by increased intraluminal pressure, flattened the endothelial cells and suppressed IP3-mediated Ca2+ signals in all activated cells. By computationally modelling Ca2+ release, endothelial shape changes were shown to alter the geometry of the Ca2+ diffusive environment near IP3 receptor microdomains to limit IP3-mediated Ca2+ signals as pressure increased. Changes in cell shape produce a geometric microdomain regulation of IP3-mediated Ca2+ signalling to explain macroscopic pressure-dependent, endothelial mechanosensing without the need for a conventional mechanoreceptor. The suppression of IP3-mediated Ca2+ signalling may explain the decrease in endothelial activity as pressure increases. GRIN imaging provides a convenient method that gives access to hundreds of endothelial cells in intact arteries in physiological configuration.

Citation

Wilson, C., Saunter, C. D., Girkin, J. M., & McCarron, J. G. (2015). Pressure-dependent regulation of Ca2+ signalling in the vascular endothelium. The Journal of Physiology, 593(24), 5231-5253. https://doi.org/10.1113/jp271157

• Journal Article Type: Article
• Acceptance Date: Oct 19, 2015
• Online Publication Date: Oct 28, 2015
• Publication Date: Dec 15, 2015
• Deposit Date: Dec 18, 2015
• Publicly Available Date: Jan 18, 2016
• Journal: Journal of Physiology
• Print ISSN: 0022-3751
• Electronic ISSN: 1469-7793
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 593
• Issue: 24
• Pages: 5231-5253
• DOI: https://doi.org/10.1113/jp271157
• Public URL: https://durham-repository.worktribe.com/output/1396032