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Am J Physiol Cell Physiol (July 16, 2003). doi:10.1152/ajpcell.00153.2003
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Submitted on April 18, 2003
Accepted on July 10, 2003

{beta}1 SUBUNITS ARE REQUIRED FOR REGULATION OF COUPLING BETWEEN Ca2+ TRANSIENTS AND Ca2+ ACTIVATED K+ (BK) CHANNELS BY PROTEIN KINASE C

Brian M Hagen1, Orline P Bayguinov1, and Kenton M Sanders1*

1 Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV, USA

* To whom correspondence should be addressed. E-mail: kent{at}physio.unr.edu.

Colonic myocytes have spontaneous, localized, IP3 receptor-dependent Ca2+ transients that couple to the activation of Ca2+-dependent K+ channels and spontaneous transient outward currents (STOCs). We previously reported that the coupling strength between spontaneous Ca2+ transients and large conductance Ca2+ activated K+ (BK) channels is regulated by Ca2+ influx through nonselective cation channels and activation of protein kinase C (PKC). Here we used confocal microscopy and the patch clamp technique to further investigate the coupling between localized Ca2+ transients and STOCs in colonic myocytes from animals lacking the regulatory {beta}1 subunit of BK channels. Myocytes from {beta}1-knockout ({beta}1-/-) animals loaded with fluo-4 showed typical localized Ca2+ transients, but the STOCs coupled to these events were of abnormally low amplitude. Reduction in external Ca2+ or application of inhibitors of nonselective cation channels (SKF-96365) caused no significant change in the amplitude or frequency of STOCs. Likewise, an inhibitor of PKC, GF-109203X, had no significant effect on STOCs. Single channel recording from BK channels showed that application of an activator (PMA) and an inhibitor (GF-109203X) of PKC did not affect BK channel openings in myocytes of {beta}1-/- mice. These data show that PKC-dependent regulation of coupling strength between Ca2+ transients and STOCs in colonic myocytes depends upon the interaction between {alpha} and {beta}1 subunits.




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