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Our recent electrophysiological analysis of mouse retinal pigment epithelial (RPE) cells revealed that in the presence of 10 mM external thiocyanate (SCN − ), voltage steps generated large transient currents whose time-dependent decay most likely results from the accumulation or depletion of SCN −  intracellularly. In the present study, we investigated the effects of more physiologically relevant concentrations of this biologically active anion. In whole cell recordings of C57BL/6J mouse RPE cells, we found that, over the range of 50 to 500 µM SCN − , the amplitude of transient currents evoked by voltage steps was proportional to the extracellular SCN −  concentration. Transient currents were also produced in RPE cells when the membrane potential was held constant and the external SCN −  concentration was rapidly increased by pressure-ejecting 500 µM SCN −  from a second pipette. Other results indicate that the time dependence of currents produced by both approaches results from a change in driving force due to intracellular SCN −  accumulation or depletion. Finally, by applying fluorescence imaging and voltage-clamping techniques to BALB/c mouse RPE cells loaded with the anion-sensitive dye MQAE, we demonstrated that in the presence of 200 or 500 µM extracellular SCN − , depolarizing voltage steps increased the cytoplasmic SCN −  concentration to an elevated steady state within several seconds. Collectively, these results indicate that, in the presence of physiological concentrations of SCN −  outside the RPE, the conductance and permeability of the RPE cell membranes for SCN −  are sufficiently large that SCN −  rapidly approaches electrochemical equilibrium within the cytoplasm when the membrane voltage or external SCN −  concentration is perturbed.

Electrophysiological impact of thiocyanate on isolated mouse retinal pigment epithelial cells