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Am J Physiol Cell Physiol 281: C1769-C1775, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 6, C1769-C1775, December 2001

Micromolar Ca2+ from sparks activates Ca2+-sensitive K+ channels in rat cerebral artery smooth muscle

Guillermo J. Pérez, Adrian D. Bonev, and Mark T. Nelson

Department of Pharmacology, University of Vermont, Burlington, Vermont 05405

The goal of the present study was to test the hypothesis that local Ca2+ release events (Ca2+ sparks) deliver high local Ca2+ concentration to activate nearby Ca2+-sensitive K+ (BK) channels in the cell membrane of arterial smooth muscle cells. Ca2+ sparks and BK channels were examined in isolated myocytes from rat cerebral arteries with laser scanning confocal microscopy and patch-clamp techniques. BK channels had an apparent dissociation constant for Ca2+ of 19 µM and a Hill coefficient of 2.9 at -40 mV. At near-physiological intracellular Ca2+ concentration ([Ca2+]i; 100 nM) and membrane potential (-40 mV), the open probability of a single BK channel was low (1.2 × 10-6). A Ca2+ spark increased BK channel activity to 18. Assuming that 1-100% of the BK channels are activated by a single Ca2+ spark, BK channel activity increases 6 × 105-fold to 6 × 103-fold, which corresponds to ~30 µM to 4 µM spark Ca2+ concentration. 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester caused the disappearance of all Ca2+ sparks while leaving the transient BK currents unchanged. Our results support the idea that Ca2+ spark sites are in close proximity to the BK channels and that local [Ca2+]i reaches micromolar levels to activate BK channels.

ryanodine receptor; local calcium release; BK potassium channel


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