The effect of pH(o) on plasma membrane chloride current of cultured rat cortical astrocytes was investigated using the whole-cell patch-clamp technique. In the presence of intra- and extracellular solutions with symmetrical high Cl(-) content and K(+) channel inhibitors, the cells exhibited an inwardly rectifying current. The current activated slowly at potentials negative to -40 mV and did not display time-dependent inactivation. The current was inhibited by 0.1 mM Cd(2+), 0.1 mM Zn(2+), 1 mM 9-anthracene-carboxylic acid, and 0.2 mM 5-nitro-2-(3-phenylpropylamino)benzoic acid, but not by 10 mM Ba(2+) or 3 mM Cs(+). Reversal potential of the current followed the chloride equilibrium potential and was not influenced by changes in K(+) or Na(+) concentration. The inwardly rectifying chloride current was augmented by extracellular acidosis and reduced by alkalosis. The pH sensitivity was most pronounced in the physiologically relevant pH(o) range of 6.9--7.9. Lowering pH to 6.4 induced no additional increase in steady-state current amplitude compared with pH(o) 6.9, but it substantially slowed the activation kinetics. According to its kinetic and pharmacological properties this chloride current is similar to that found in cultured rat astrocytes after long-term treatment with dibutyryl-cAMP, however, in our cultures it was consistently expressed without any treatment with the drug. Considering that astrocytes possess carbonic anhydrase and Cl(-)/HCO3(-) antiporter, this current may participate in the regulation of the interstitial and astrocyte pH.