Effect of bicarbonate, pH, methazolamide and stilbenes on the intracellular potentials of cultured bovine corneal endothelial cells

Abstract

Micropuncture of cultured bovine corneal endothelial cells led to registrations stable for hours. Intracellular potentials were mainly in the range of -40 to -55 mV, average 46.3 +/- 0.6 mV (SEM). Changes of extracellular [HCO-3] led to voltage transients, their amplitude depending logarithmically on [HCO-3] with a mean slope of 37.3 +/- 8.8 (SD) mV. After removal of bicarbonate/CO2, a steady-state depolarization was seen. This steady-state depolarization, but not the voltage transients, could be reduced by 1 mM Ba++. After removal of bicarbonate, the voltage response to changes of extracellular potassium was reduced. Alteration of pHi induced by permeable buffers (butyrate, glycodiazine and ammonium) also resulted in voltage transients, internal acidification being correlated with a hyperpolarization, and internal alkalinization with a depolarization. Also changes of external pH caused voltage responses, alkalinization causing a hyperpolarization, acidification a depolarization. Methazolamide, an inhibitor of carbonic anhydrase, as well as stilbenes (SITS or DIDS) caused a reduction of the voltage response to HCO-3 and pH. Their effects were additive. It is suggested that corneal endothelial cells possess one or two electrogenic transporters for HCO-3 or related species, one of which is inhibitable by stilbenes.