We have previously reported that angiotensin II (ANG II) induces oscillations in the cytoplasmic calcium concentration ([Ca2+](i)) of pulmonary vascular myocytes. The present work was undertaken to investigate the effect of ANG II in comparison with ATP and caffeine on membrane currents and to explore the relation between these membrane currents and [Ca2+](i). In cells clamped at -60 mV, ANG 11 (10 mu M) or ATP (100 mu M) induced an oscillatory inward current. Caffeine (5 mM) induced only one transient inward current. In control conditions, the reversal potential (E-rev) Of these currents was close to the equilibrium potential for Cl- ions (E-Cl = -2.1 mV) and was shifted towards more positive values in low-Cl- solutions. Niflumic acid (10-50 mu M) and DIDS (0.25-1 mM) inhibited this inward current. Combined recordings of membrane current and [Ca2+](i) by Indo-1 microspectrofluorimetry revealed that ANG II- and ATP-induced currents occurred simultaneously with oscillations in [Ca2+](i), whereas the caffeine-induced current was accompanied by only one transient increase in [Ca2+](i). Niflumic acid (25 mu M) had no effect on agonist-induced [Ca2+](i) responses, whereas thapsigargin (1 mu M) abolished both membrane current and the [Ca2+](i) response. Heparin (5 mg/ml in the pipette solution) inhibited both [Ca2+](i) responses and membrane currents induced by ANG II and ATP, but not by caffeine. In pulmonary arterial strips, ANG It-induced contraction was inhibited by niflumic acid (25 mu M) or nifedipine (1 mu M) to the same extent and the two substances did not have an additive effect. This study demonstrates that, in pulmonary vascular smooth muscle, ANG II, as well as ATP, activate an oscillatory calcium dependent chloride current which is triggered by cyclic increases in [Ca2+](i) and that both oscillatory phenomena are primarily IP3-mediated. It is suggested that ANG Ii-induced oscillatory chloride current could depolarise the cell membrane leading to activation of voltage-operated Ca2+ channels. The resulting Ca2+ influx contributes to the component of ANG Ii-induced contraction that is equally sensitive to chloride or calcium channel blockade.
