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Division of Pulmonary and Critical Care Medicine, Department of Medicine, The Johns Hopkins University, Baltimore, Maryland 21224
Endothelin-1 (ET-1)
increases intracellular Ca2+ concentration
([Ca2+]i) in pulmonary arterial
smooth muscle cells (PASMCs); however, the mechanisms for
Ca2+ mobilization are not clear. We determined the
contributions of extracellular influx and intracellular release to the
ET-1-induced Ca2+ response using Indo 1 fluorescence and
electrophysiological techniques. Application of ET-1
(10
10 to 108 M) to transiently
(24-48 h) cultured rat PASMCs caused concentration-dependent increases in [Ca2+]i. At
108 M, ET-1 caused a large, transient increase in
[Ca2+]i (>1 µM) followed by a
sustained elevation in [Ca2+]i
(<200 nM). The ET-1-induced increase in
[Ca2+]i was attenuated (<80%) by
extracellular Ca2+ removal; by verapamil, a voltage-gated
Ca2+-channel antagonist; and by ryanodine, an inhibitor of
Ca2+ release from caffeine-sensitive stores. Depleting
intracellular stores with thapsigargin abolished the peak in
[Ca2+]i, but the sustained phase
was unaffected. Simultaneously measuring membrane potential and
[Ca2+]i indicated that
depolarization preceded the rise in
[Ca2+]i. These results suggest that
ET-1 initiates depolarization in PASMCs, leading to Ca2+
influx through voltage-gated Ca2+ channels and
Ca2+ release from ryanodine- and inositol
1,4,5-trisphosphate-sensitive stores.
calcium-induced calcium release; vascular smooth muscle; potassium chloride; L-type calcium channels
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