Sorcin modulation of Ca2+ sparks in rat vascular smooth muscle cells.

Spontaneous, local Ca(2+) release events or Ca(2+) sparks by ryanodine receptors (RyRs) are important determinants of vascular tone and arteriolar resistance, but the mechanisms that modulate their properties in smooth muscle are poorly understood. Sorcin, a Ca(2+)-binding protein that associates with cardiac RyRs and quickly stops Ca(2+) release in the heart, provides a potential mechanism to modulate Ca(2+) sparks in vascular smooth muscle, but little is known about the functional role of sorcin in this tissue. In this work, we characterized the expression and intracellular location of sorcin in aorta and cerebral artery and gained mechanistic insights into its functional role as a modulator of Ca(2+) sparks. Sorcin is present in endothelial and smooth muscle cells, as assessed by immunocytochemical and Western blot analyses. Smooth muscle sorcin translocates from cytosolic to membranous compartments in a Ca(2+)-dependent manner and associates with RyRs, as shown by coimmunoprecipitation and immunostaining experiments. Ca(2+) sparks recorded in saponin-permeabilized vascular myocytes have increased frequency, duration and spatial spread but reduced amplitude with respect to Ca(2+) sparks in intact cells, suggesting that permeabilization disrupts the normal organization of RyRs and releases diffusible substances that control Ca(2+) spark properties. Perfusion of 2 mum sorcin onto permeabilized myocytes reduced the amplitude, duration and spatial spread of Ca(2+) sparks, demonstrating that sorcin effectively regulates Ca(2+) signalling in vascular smooth muscle. Together with a dense distribution in the perimeter of the cell along a pool of RyRs, these properties make sorcin a viable candidate to modulate vascular tone in smooth muscle.