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Am J Physiol Heart Circ Physiol 293: H204-H214, 2007. First published March 2, 2007; doi:10.1152/ajpheart.00669.2006
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Caveolin-1 and caveolin-3 regulate Ca2+ homeostasis of single smooth muscle cells from rat cerebral resistance arteries

T. Kamishima,1 T. Burdyga,2 J. A. Gallagher,1 and J. M. Quayle1

Departments of 1Human Anatomy and Cell Biology and 2Physiology, School of Biomedical Sciences, University of Liverpool, Liverpool, United Kingdom

Submitted 23 June 2006 ; accepted in final form 27 February 2007

The role of caveolins, signature proteins of caveolae, in arterial Ca2+ regulation is unknown. We investigated modulation of Ca2+ homeostasis by caveolin-1 and caveolin-3 using smooth muscle cells from rat cerebral resistance arteries. Membrane current and Ca2+ transients were simultaneously measured with voltage-clamped single cells. Membrane depolarization triggered Ca2+ current and increased intracellular Ca2+ concentration ([Ca2+]i). After repolarization, elevated [Ca2+]i returned to the resting level. Ca2+ removal rate was determined from the declining phase of the Ca2+ transient. Application of caveolin-1 antibody or caveolin-1 scaffolding domain peptide, corresponding to amino acid residues 82–101 of caveolin-1, significantly slowed Ca2+ removal rate at a measured [Ca2+]i of 250 nM, with little effect at a measured [Ca2+]i of 600 nM. Application of caveolin-3 antibody or caveolin-3 scaffolding domain peptide, corresponding to amino acid residues 55–74 of caveolin-3, also significantly slowed Ca2+ removal rate at a measured [Ca2+]i of 250 nM, with little effect at a measured [Ca2+]i of 600 nM. Likewise, application of calmodulin inhibitory peptide, autocamtide-2-related inhibitory peptide, and cyclosporine A, inhibitors for calmodulin, Ca2+/calmodulin-dependent protein kinase II, and calcineurin, also significantly inhibited Ca2+ removal rate at a measured [Ca2+]i of 250 nM but not at 600 nM. Application of cyclopiazonic acid, a sarcoplasmic reticulum Ca2+ ATPase inhibitor, also significantly inhibited Ca2+ removal rate at a measured [Ca2+]i of 250 nM but not at 600 nM. Our results suggest that caveolin-1 and caveolin-3 are important in Ca2+ removal of resistance artery smooth muscle cells.

caveolin-1 scaffolding domain peptide; caveolin-3 scaffolding domain peptide; calmodulin; calcium/calmodulin-dependent protein kinase II; calcineurin



Address for reprint requests and other correspondence: J. M. Quayle, Dept. of Human Anatomy and Cell Biology, School of Biomedical Sciences, Univ. of Liverpool, The Sherrington Bldgs., Ashton St., Liverpool L69 3GE, UK (e-mail: jquayle{at}liv.ac.uk)




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