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Received August 1, 2001
Accepted after revision October 15, 2001
1 Department of Pharmacology and Clinical Pharmacology, St George's Hospital Medical School, Cranmer Terrace, London SW17 ORE, UK
* To whom correspondence should be addressed. E-mail: largew{at}sghms.ac.uk.
In vascular smooth muscle cells, many agonists cause the release of Ca2+ ions from internal stores. An important problem concerns the mechanism by which the intracellular stores are refilled subsequent to depletion. In the present study, we describe the properties of a Ca2+-permeable non-selective cation channel current that is activated in rabbit portal vein myocytes by depletion of internal Ca2+ stores. Application of cyclopiazonic acid (CPA), which depletes internal Ca2+ stores, activated whole-cell currents that had a reversal potential (Er) of about +50 mV in 1.5 m external Ca2+ (Ca). In 0 mm Ca, the currents were larger and Er was ~0 mV. Application of CPA and caffeine during cell-attached recording activated single inward channel currents at negative potentials, which had a slope conductance of 2-3 pS and an Er of +20 mV. The slope conductance in 0 and 110 mm Ca was 7 and 1.5 pS, respectively, and Er values indicated that these non-selective cation channels are highly permeable to Ca2+ ions. Bath application of the cell-permeant Ca2+ chelator, BAPTA-AM, also activated similar currents, indicating that these channels are not activated by Ca2+. Spontaneous channel currents with similar properties to store-operated channels were observed in some patches. Application of W-7, an inhibitor of the Ca2+-binding protein, calmodulin, also activated similar Ca2+-permeable channel currents. In conclusion, it is demonstrated that agents that deplete Ca2+ stores and inhibit calmodulin binding activate Ca2+-permeable non-selective cation channel currents in rabbit portal vein myocytes. These channels might play an important role in vascular smooth muscle in facilitating an influx of Ca2+ to refill depleted internal Ca2+ stores and appear to possess different characteristics to store-operated channels.
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