Protein kinase C stimulates the small-conductance K+ channel in the basolateral membrane of the CCD

被引:14
|
作者
Lu, M [1 ]
Wang, WH [1 ]
机构
[1] NEW YORK MED COLL, DEPT PHARMACOL, VALHALLA, NY 10595 USA
关键词
potassium channel; nitric oxide; guanosine; 3'; 5'-cyclic monophosphate; collecting duct; rat kidney;
D O I
10.1152/ajprenal.1996.271.5.F1045
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
We have used the patch-clamp technique to study the regulation of the activity of the basolateral small-conductance K+ channel (SK) in the cortical collecting duct (CCD) of the rat kidney. Addition of 50-75 nM calphostin C, an agent which specifically inhibits protein kinase C (PKC), reduced channel activity by 90% in cell-attached patches. In contrast, addition of 1 mu M phorbol 12-myristate 13-acetate, a stimulator of PKC, led to addition of ''new'' K+ channel currents in 9 of 20 patches in the basolateral membrane of the CCD, and the mean increase in NPo, a product of channel number (N) and open probability (P-o), was 0.90 in these 9 patches. However, application of 1 nM exogenous PKC had no significant effect on channel activity in inside-out patches, suggesting that the PKC effect on the activity of the SK observed in cell-attached patches was not a result of a membrane-delimited action, such as a direct phosphorylation of the SK or closely associated proteins. The effect of calphostin C on the SK can be reversed by addition of either 10 mu M S-nitroso-N-acetylpenicillamine, a donor of nitric oxide, or 100 mu M 8-bromoguanosine 3',5'-cyclic monophosphate. In addition, the inhibitory effect of calphostin C on the SK was completely abolished by pretreatment of the cells with 1 mu M okadaic acid, an inhibitor of protein phosphatase. However, 100 mu M N-omega-nitro-L-arginine methyl ester, an agent that inhibits nitric oxide synthases (NOS), blocked the SK in cell-attached patches in the presence of okadaic acid, suggesting that the effect of okadaic acid on calphostin C-induced inhibition of the SK was a step before formation of nitric oxide. We conclude that PKC is involved in the stimulation of the SK and that the effect of PKC on the SK may be mediated by regulation of NOS activity in the CCD of the rat kidney.
引用
收藏
页码:F1045 / F1051
页数:7
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