Heterogeneity in cytosolic calcium regulation among different microvascular smooth muscle cells of the rat retina.

Rat retinae were dissociated to yield intact microvessels 7 to 42 microm in diameter. These were loaded with fura-2 AM and single fragments anchored down in a recording bath. Intracellular Ca(2+) levels from 20- to 30-microm sections of vessel were estimated by microfluorimetry. The vessels studied were identified as metarterioles and arterioles. Only the microvascular smooth muscle cells loaded with fura-2 AM and changes in the fluorescence signal were confined to these cells: Endothelial cells did not make any contribution to the fluorescence signal nor did they contribute to the actions of the drugs. Caffeine (10 mM) or elevated K(+) (100 mM) produced a transient rise in cell Ca(2+) in the larger vessels (diameters >18 microm) but had no effect on smaller vessels (diameters <18 microm). Rises in cell Ca(2+) were accompanied by a rapid ( approximately 2 s to peak) contraction followed by relaxation. Caffeine and K(+) responses were blocked by ryanodine (10 microM) and nifedipine (1 microM), respectively. In all the vessels tested, vasopressin (arginine, 10 nM) elicited a transient increase in cell Ca(2+) and a constriction, irrespective of the diameter of the vessel. All vessels tested also responded to endothelin-1 (1-10 nM) through an Et(A) receptor to produce a transient rise in cell Ca(2+) followed by a plateau phase of elevated Ca(2+) and a constriction. In contrast to the transient effects of vasopressin, caffeine, and K(+), the cell Ca(2+) remained elevated (>30 min) on washing out the endothelin and the vessel failed to relax. These results demonstrate heterogeneity between smaller and larger retinal vessels with regard to Ca(2+) mobilisation and homogeneity with respect to the actions of vasoactive peptides. Copyright 2000 Academic Press.