The effects of beta-stimulation on the Na(+)-K+ pump current-voltage relationship in guinea-pig ventricular myocytes.

1. The whole cell patch clamp technique was used to study effects of the beta agonist isoprenaline (Iso) on the current-voltage (I-V) relationship of the Na(+)-K+ pump current (Ip) in acutely isolated guinea-pig ventricular myocytes. 2. The effect of Iso on Ip at high [Ca2+]i (1.4 microM) was voltage dependent. The I-V relationship of Ip in Iso shifted by approximately 30 mV in the negative direction on the voltage axis, increasing Ip at negative voltages but leaving Ip unchanged at positive voltages. 3. Intracellular application of the calmodulin antagonist, calmodulin-dependent protein kinase II fragment 290-309, did not eliminate or reduce the Iso-induced voltage shift, suggesting calmodulin-dependent protein kinase II was not involved. 4. The Iso inhibition of Ip at low [Ca2+]i (15 nM) was not voltage dependent. Ip was reduced by 20 to 30% in the presence of Iso at each holding potential. 5. When the voltage dependence of Ip was largely reduced by substitution of N-methyl-D-glucamine+ for external Na+, the magnitude of the low [Ca2+]i, Iso-induced inhibition of Ip was progressively eliminated by increasing the [Ca2+]i. At a [Ca2+]i of 1.4 microM, this inhibition disappeared. 6. At intermediate values of [Ca2+]i, the I-V curves in Na(+)-containing solution in the presence and the absence of Iso crossed over. The higher the [Ca2+]i, the more positive the voltage at which the two I-V curves intersected. 7. During beta-adrenergic activation our results suggest intracellular Ca2+ has two effects: (a) It prevents protein kinase A (PKA) phosphorylation-induced inhibition of Ip. (b) It causes a PKA phosphorylation-induced shift of the pump I-V relationship in the negative direction on the voltage axis. These effects may have important physiological significance in the regulation of heart rate and cardiac contractility.