Nitric oxide stimulates a large-conductance Ca2+-activated K+ channel in human skin fibroblasts through protein kinase G pathway

초록

In order to investigate the large-conductance Ca2+- activated K+ (BKCa) channel and determine the effects of nitric oxide (NO) on the channel in human skin fibroblasts, we performed electrophysiological patch clamp recordings on 5th-passage cells of human genital skin cultures. The whole-cell outward K+ current was increased with depolarization, and proved to be sensitive to NS1619 ( a selective BKCa channel activator) and iberiotoxin ( a specific BKCa channel inhibitor). The single-channel currents showed 226 pS of mean conductance in symmetrical K+. Sodium nitroprusside ( SNP; an NO donor) significantly increased the K+ current amplitude in the whole-cell mode, and open probability of the channel (NPo) in the cell-attached mode, but not in the inside-out mode. S-nitroso-N- acetylpenicillamine ( an NO donor) and 8-Br-cGMP ( a membrane-permeant cGMP analogue) also increased the BKCa channel activity. The stimulatory effect of SNP on BKCa channels was inhibited by pretreatment with 1H-[1,2,4]- oxadiazolo[4,3-a] quinoxalin-1-one (a soluble guanylyl cyclase inhibitor), or KT5823 [ a specific protein kinase G (PKG) inhibitor]. Cytoplasmic PKG also increased the channel activity in inside-out patches. In conclusion, the present data indicate that BKCa channels constitute a significant fraction of K+ current in human skin fibroblasts, and that NO increases NPo of BKCa channels, which are mediated via the cGMP/PKG pathway, without direct effects on the channel. Copyright (C) 2005 S. Karger AG, Basel.

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