Sodium-calcium exchanger contributes to membrane hyperpolarization of intact endothelial cells from rat aorta during acetylcholine stimulation

1 The role of sodium - calcium exchanger in acetylcholine (Ach)-induced hyperpolarization of intact endothelial cells was studied in excised rat aorta. The membrane potential was recorded using perforated patch-clamp technique. 2 The mean resting potential of endothelial cells was - 44.1 +/- 1.4 mV. A selective inhibitor of sodium - calcium exchanger benzamil ( 100 muM) had no significant effect on resting membrane potential, but reversibly decreased the amplitude of sustained Ach-induced endothelial hyperpolarization from 20.9 +/- 1.4 to 5.7 +/- 1.1 mV when applied during the plateau phase. 3 The blocker of reversed mode of the exchanger KB-R7943 (2-[2-[4-(4- nitrobenzyloxy) phenyl] ethyl] isothiourea methanesulfonate, 20 mM) reversibly decreased the amplitude of sustained Ach-induced hyperpolarization from 20.5 +/- 2.9 to 7.5 +/- 1.8 mV. 4 Introduction of tetraethylammonium (10 mM) in the continuous presence of Ach decreased the sustained phase of hyperpolarization from 17.9 +/- 1.5 by 12.9 +/- 0.9 mV. Subsequent addition of 20 mM KB-R7943 further depolarized endothelial cells by 4.8 +/- 1.1 mV. 5 Substituting external sodium with N-methyl D-glucamine during the plateau phase of Ach-evoked hyperpolarization reversibly decreased the hyperpolarization from - 61.8 +/- 2.7 to -54.2 +/- 1.9 mV. In the majority of preparations, the initial response to removal of external sodium was a transient further rise in the membrane potential of several mV. Sodium ionophore monensin hyperpolarized endothelium by 10.3 +/- 0.7 mV. 6 The inhibitory effect of benzamil on Ach-induced endothelial sustained hyperpolarization was observed in endothelium mechanically isolated from smooth muscle. 7 These results suggest that the sodium - calcium exchanger of intact endothelial cells is able to operate in reverse following stimulation by Ach, contributing to sustained hyperpolarization. Myoendothelial electrical communications do not mediate the effect of blockers of sodium - calcium exchanger.