Regulation of membrane excitability by intracellular pH (pH i ) changers through Ca 2+ -activated K + current (BK channel) in single smooth muscle cells from rabbit basilar artery

Employing microfluorometric system and patch clamp technique in rabbit basilar arterial myocytes, regulation mechanisms of vascular excitability were investigated by applying intracellular pH (pH i ) changers such as sodium acetate (SA) and NH 4 Cl. Applications of caffeine produced transient phasic contractions in a reversible manner. These caffeine-induced contractions were significantly enhanced by SA and suppressed by NH 4 Cl. Intracellular Ca 2+ concentration ([Ca 2+ ] i ) was monitored in a single isolated myocyte and based the ratio of fluorescence using Fura-2 AM ( R 340/380 ). SA (20 mM) increased and NH 4 Cl (20 mM) decreased R 340/380 by 0.2 ± 0.03 and 0.1 ± 0.02, respectively, in a reversible manner. Caffeine (10 mM) transiently increased R 340/380 by 0.9 ± 0.07, and the ratio increment was significantly enhanced by SA and suppressed by NH 4 Cl, implying that SA and NH 4 Cl may affect [Ca 2+ ] i ( p < 0.05). Accordingly, we studied the effects of SA and NH 4 Cl on Ca 2+ -activated K + current ( I K Ca ) under patch clamp technique. Caffeine produced transient outward current at holding potential ( V h ) of 0 mV, caffeine induced transient outward K + current, and the spontaneous transient outward currents were significantly enhanced by SA and suppressed by NH 4 Cl. In addition, I K Ca was significantly increased by acidotic condition when pH i was lowered by altering the NH 4 Cl gradient across the cell membrane. Finally, the effects of SA and NH 4 Cl on the membrane excitability and basal tension were studied: Under current clamp mode, resting membrane potential (RMP) was −28 ± 2.3 mV in a single cell level and was depolarized by 13 ± 2.4 mV with 2 mM tetraethylammonium (TEA). SA hyperpolarized and NH 4 Cl depolarized RMP by 10 ± 1.9 and 16 ± 4.7 mV, respectively. SA-induced hyperpolarization and relaxation of basal tension was significantly inhibited by TEA. These results suggest that SA and NH 4 Cl might regulate vascular tone by altering membrane excitability through modulation of [Ca 2+ ] i and Ca 2+ -activated K channels in rabbit basilar artery.