Different effects of endothelin-1 on calcium and potassium currents in canine ventricular cells

Abstract.

Effects of endothelin-1 (ET-1) on the L-type calcium current (I _Ca) and delayed rectifier potassium current (I _K) were studied in isolated canine ventricular cardiomyocytes using the whole-cell configuration of the patch-clamp technique. ET-1 (8 nM) was applied in three experimental arrangements: untreated cells, in the presence of 50 nM isoproterenol, and in the presence of 250 µM 8-bromo-cAMP. In untreated cells, ET-1 significantly decreased the peak amplitude of I _Ca by 32.3±4.8% at +5 mV (P<0.05) without changing activation or inactivation characteristics of I _Ca. ET-1 had no effect on the amplitude of I _K, I _to (transient outward current) or I _K1 (inward rectifier K current) in untreated cells; however, the time course of recovery from inactivation of I _to was significantly increased by ET-1 (from 26.5±4.6 ms to 59.5±1.8 ms, P<0.05). Amplitude and time course of intracellular calcium transients, recorded in voltage-clamped cells previously loaded with the fluorescent calcium indicator dye Fura-2, were not affected by ET-1. ET-1 had no effect on force of contraction in canine ventricular trabeculae.

Isoproterenol increased the amplitude of I _Ca to 263±29% of control. ET-1 reduced I _Ca also in isoproterenol-treated cells by 17.8±2% (P<0.05); this inhibition was significantly less than obtained in untreated cells. I _K was increased by isoproterenol to 213±18% of control. This effect of isoproterenol on I _K was reduced by 31.8±4.8% if the cells were pretreated with ET-1. Similarly, in isoproterenol-treated cells ET-1 decreased I _K by 16.2±1.5% (P<0.05). Maximal activation of protein kinase A (PKA) was achieved by application of 8-bromo-cAMP in the pipette solution. In the presence of 8-bromo-cAMP ET-1 failed to alter I _Ca or I _K. It was concluded that differences in effects of ET-1 on I _Ca and I _K may be related to differences in cAMP sensitivity of the currents.