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1 Department of Pharmacology, University of Nevada School of Medicine, Reno, NV 89557, USA
Capacitative Ca2+ entry (CCE) has been speculated to contribute to Ca2+ influx during hypoxic pulmonary vasoconstriction (HPV). The aim of the present study was to directly test if acute hypoxia causes intracellular Ca2+ concentration ([Ca2+]i) rises through CCE in canine pulmonary artery smooth muscle cells (PASMCs). In PASMCs loaded with fura-2, hypoxia produced a transient rise in [Ca2+]i in Ca2+-free solution, indicating Ca2+ release from the intracellular Ca2+ stores. Subsequent addition of 2 mM Ca2+ in hypoxia elicited a sustained rise in [Ca2+]i, which was partially inhibited by 10 µM nisoldipine. The dihydropyridine-insensitive rise in [Ca2+]i was due to increased Ca2+ influx, because it was abolished in Ca2+-free solution and hypoxia was shown to significantly enhance the rate of Mn2+ quench of fura-2 fluorescence. The dihyropyridine-insensitive rise in [Ca2+]i and the increased rate of Mn2+ quench of fura-2 fluorescence were inhibited by 50 µM SKF 96365 and 500 µM Ni2+, but not by 100 µM La3+ or 100 µM Gd3+, exhibiting pharmacological properties characteristic of CCE. In addition, predepletion of the intracellular Ca2+ stores inhibited the rise in [Ca2+]i induced by hypoxia. These results provide the first direct evidence that acute hypoxia, by causing Ca2+ release from the intracellular stores, activates CCE in isolated canine PASMCs, which may contribute to HPV.
(Received 29 October 2004;
accepted after revision 20 December 2004;
first published online 21 December 2004)
Corresponding author J. R. Hume: Department of Pharmacology/318, University of Nevada School of Medicine, Reno, NV 89557, USA. Email: joeh{at}med.unr.edu
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