Effects of a protein phosphatase inhibitor, okadaic acid, on membrane currents of isolated guinea-pig cardiac myocytes

Abstract

The effects of a protein phosphatase inhibitor, okadaic acid (OA), were studied on membrane currents of isolated myocytes from guinea-pig cardiac ventricle. The whole-cell Ca2+ current (ICa) was recorded as peak inward current in response to test pulse to 0 mV. Extracellular application of OA (5-100 microM) produced an increase of ICa. The effect was markedly enhanced when the myocyte was pretreated with threshold concentrations of isoprenaline. ICa was increased from 11.3 +/- 0.8 microA cm-2 to 19.0 +/- 1.1 microA cm-2 (n = 4) by 5 microM-OA in the presence of 1 nM-isoprenaline. The delayed rectifier current was also slightly increased. Furthermore, the wash-out time of the beta-adrenergic increase of ICa was markedly prolonged by OA. The beta-adrenergic stimulation of cardiac Ca2+ current is thought to be mediated by cAMP-dependent phosphorylation. The present results strongly suggest that the effect of OA on ICa is related to inhibition of endogenous protein phosphatase activity which is responsible for the dephosphorylation process. By the isotope method, the inhibitory effect of OA on different types of phosphatase was compared. OA had a relatively high specificity to type 1-, and type 2A-phosphatases.