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Am J Physiol Cell Physiol 280: C22-C33, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 1, C22-C33, January 2001

Heterogeneity of calcium stores and elementary release events in canine pulmonary arterial smooth muscle cells

Robert Janiak, Sean M. Wilson, Stephen Montague, and Joseph R. Hume

Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada 89557

To examine the nature of inositol 1,4,5-trisphosphate (IP3)-sensitive and ryanodine (Ryn)-sensitive Ca2+ stores in isolated canine pulmonary arterial smooth cells (PASMC), agonist-induced changes in global intracellular Ca2+ concentration ([Ca2+]i) were measured using fura 2-AM fluorescence. Properties of elementary local Ca2+ release events were characterized using fluo 3-AM or fluo 4-AM, in combination with confocal laser scanning microscopy. In PASMC, depletion of sarcoplasmic reticulum Ca2+ stores with Ryn (300 µM) and caffeine (Caf; 10 mM) eliminated subsequent Caf-induced intracellular Ca2+ transients but had little or no effect on the initial IP3-mediated intracellular Ca2+ transient induced by ANG II (1 µM). Cyclopiazonic acid (CPA; 10 µM) abolished IP3-induced intracellular Ca2+ transients but failed to attenuate the initial Caf-induced intracellular Ca2+ transient. These results suggest that in canine PASMC, IP3-, and Ryn-sensitive Ca2+ stores are organized into spatially distinct compartments while similar experiments in canine renal arterial smooth muscle cells (RASMC) reveal that these Ca2+ stores are spatially conjoined. In PASMC, spontaneous local intracellular Ca2+ transients sensitive to modulation by Caf and Ryn were detected, exhibiting spatial-temporal characteristics similar to those previously described for "Ca2+ sparks" in cardiac and other types of smooth muscle cells. After depletion of Ryn-sensitive Ca2+ stores, ANG II (8 nM) induced slow, sustained [Ca2+]i increases originating at sites near the cell surface, which were abolished by depleting IP3 stores. Discrete quantal-like events expected due to the coordinated opening of IP3 receptor clusters ("Ca2+ puffs") were not observed. These data provide new information regarding the functional properties and organization of intracellular Ca2+ stores and elementary Ca2+ release events in isolated PASMC.

pulmonary artery; smooth muscle cells; sarcoplasmic reticulum; intracellular calcium; inositol 1,4,5-trisphosphate; calcium ion sparks


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