Responsiveness of cytosolic free calcium concentration in cultured rat pulmonary arterial smooth muscle cells: confocal microscopic measurement.

Rat pulmonary arterial smooth muscle cells were primarily cultured. alpha-actins in pulmonary arterial smooth muscle cells at day 7 of culture were observed by an immunofluorescence staining method using anti-alpha-actin antibody. Microfluorimetry of Ca2+ signals in fluo-3-loaded single smooth cell at day 7 of culture was performed by a laser-scanned confocal imaging system. The effects of several kinds of Ca(2+)-mobilizing drugs on cytoplasmic Ca2+ concentration ([Ca2+]i) were examined. KCl, a depolarizing agent, and norepinephrine, an alpha-adrenergic agonist, equally increased [Ca2+]i. Angiotensin II, a receptor agonist, and caffeine, a Ca(2+)-induced Ca2+ releaser, elevated [Ca2+]i in the same manner but was more potent than KCl and norepinephrine. Br-A23187, a Ca2+ ionophore, most potently increased [Ca2+]i. The present results suggest that drug receptors on plasma membrane, Ca2+ entry pathways and Ca(2+)-releasing mechanisms act normally, and that our cultured pulmonary arterial smooth muscle cells may be a good model for the study on the essential role of Ca2+ in vasoconstriction.