title: Protein kinase C stimulates the small-conductance K+ channel in the basolateral membrane of the CCD.
abstract: 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 microM 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 NP0, a product of channel number (N) and open probability (Pzero), 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 microM S-nitroso-N-acetylpenicillamine, a donor of nitric oxide, or 100 microM 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 microM okadaic acid, an inhibitor of protein phosphatase. However, 100 microM 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.
authors.affiliation: Department of Pharmacology, New York Medical College, Valhalla 10595, USA.
authors.forename: M
authors.initials: M
authors.lastname: Lu
authors.forename: W
authors.initials: W
authors.lastname: Wang
mesh_terms: D000818:Animals; D006152:Cyclic GMP / Q000494:pharmacology; D004553:Electric Conductivity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D007685:Kidney Tubules, Collecting / Q000378:metabolism*; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009281:Naphthalenes / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects* / Q000502:physiology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D009281:Naphthalenes; D015221:Potassium Channels; D009569:Nitric Oxide; D011493:Protein Kinase C; D006152:Cyclic GMP; C058819:calphostin C; D013755:Tetradecanoylphorbol Acetate; D019331:NG-Nitroarginine Methyl Ester