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Articles in PresS, published online ahead of print September 11, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00132.2002
Submitted on March 20, 2002
Accepted on September 4, 2002
1 Department of Cell Physiology, National Institute for Physiological Sciences, Okazaki, Aichi, Japan; Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Okazaki, Aichi, Japan
* To whom correspondence should be addressed. E-mail: okada{at}nips.ac.jp.
Parallel activation of Ca2+-dependent K+ channels and volume-sensitive Cl- channels is known to be responsible for KCl efflux during regulatory volume decrease (RVD) in human epithelial Intestine 407 cells. The present study was performed to identify the K+ channel type. RT-PCR demonstrated mRNA expression of Ca2+-activated, intermediate-conductance K+ (IK), but not small-conductance K+ (SK1) or large-conductance K+ (BK) channels in this cell line. Whole-cell recordings showed that ionomycin or hypotonic stress activated inwardly rectifying K+ currents which were reversibly blocked by IK channel blockers (clotrimazole and charybdotoxin) but not by SK and BK channel blockers (apamin and iberiotoxin). Inside-out recordings revealed the existence of clotrimazole-sensitive single K+ channel activity which exhibited an intermediate unitary conductance (30 pS at -100 mV). The channel was activated by cytosolic Ca2+ in inside-out patches and by a hypotonic challenge in cell-attached patches. The RVD was suppressed by clotrimazole, but not by apamin or iberiotoxin. Thus, we conclude that the IK channel is involved in the RVD process in these human epithelial cells.
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