Electrophysiologic techniques were used to characterize the electrical properties and the ion transport mechanisms at the apical and basolateral membranes of the human retinal pigment epithelium (RPE). These experiments used fresh native tissue from adult donor and fetal eyes. In the upper range, adult donor RPE had an apical membrane resting potential (VA) of approximately equal to -60 mV and a transepithelial potential (TEP) and resistance (Rt) of 3.5 mV and 148 omega.cm2, respectively. The means were at least 50% of these values. In RPE from fetuses of gestational age 19-23 wk, VA was -56 +/- 4 mV, TEP was 2.2 +/- 1.5 mV, Rt was 206 +/- 151 omega.cm2, and the ratio of apical to basolateral resistance was 0.70 +/- 0.50 (mean +/- standard deviation; n = 15). The apical membrane of the adult donor and fetal RPE contains a large relative K+ conductance (TK > 0.3) that is barium blockable. In fetal RPE, there is evidence for separate K+ and Cl- conductive mechanisms at the basolateral membrane. However, the evidence for the Cl- conductance is indirect. The fetal RPE apical membrane, but not the basolateral membrane, contains a ouabain-sensitive mechanism that exhibits two distinct phases of apical depolarization. The first, rapid phase suggests that the pump is electrogenic. The apical membrane of fetal RPE contains a bumetanide-sensitive mechanism and a receptor activated by nanomolar amounts of epinephrine. In fetal RPE, step changes in apical [K+]o between 5 and 2 mmol/l produced a delayed basolateral membrane hyperpolarization that in situ generates the fast oscillation trough of the electroretinogram.