Abstract
The regulatory effects of cyclic GMP on purinoceptor-operated cytoplasmic Ca2+ oscillation of rat megakaryocytes were investigated by using whole-cell patch-clamp technique. ATP-induced oscillatory K+ currents though Ca2+-activated K+ channels (I(KCa)S) were depressed by pretreatment with the guanylate cyclase activator, sodium nitroprusside, and a stable membrane-permeable cGMP analogue, 8-bromo-cGMP. The inhibition by sodium nitroprusside was blocked by treatment with a cyclic nucleotide-dependent protein kinase inhibitor, N-[2-(methylamino)]-5-isoquinolinesulfonamide x HCl (H-8) (10 microM), but not by a selective cAMP-dependent-protein kinase inhibitor, Rp-cAMPS (100 microM). The oscillatory I(KCa) directly evoked by intracellular D-myo-inositol-trisphosphate (IP3) perfusion was also inhibited by the application of sodium nitroprusside. The inhibitory effect of sodium nitroprusside disappeared when the ATP-induced oscillatory I(KCa) was changed to a monophasic sustained I(KCa) current by inhibition of Ca2+-ATPase. These results suggested that cGMP depressed Ca2+ mobilization by improving Ca2+-ATPase activity by phosphorylation.
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