Ionic permeabilities of the plasma membrane of isolated intact bovine rod outer segments as studied with a novel optical probe

Abstract

The permeability properties of the plasma membrane of intact rod outer segments purified from bovine retinas (ROS) were studied with the aid of the optical probe neutral red as described in the companion paper. The following observations were made: Electrical shunting of ROS membranes greatly stimulated Na+ and K+ transport, suggesting that this transport reflects Na+ and K+ currents, respectively. The dissipation of a Na+ gradient across the plasma membrane occurred with a half-time of 30 sec at 25 degrees C. The Na+ permeability was progressively inhibited when the external Ca2+ concentration was raised from 1 microM to 20 mM. A similar Ca2+ dependence was observed for H+ and Li+ transport. The Na+ permeability was not affected when the total internal Ca2+ content of ROS was varied between 0.1 mol Ca2+/mol rhodopsin and 7 mol Ca2+/mol rhodopsin, or when the free internal Ca2+ concentration was varied between 0.1 and 50 microM. The K+ permeability was progressively stimulated when the external Ca2+ concentration was raised from 0.001 to 1 microM, whereas a further increase to 20 mM was without effect. A similar Ca2+ dependence was observed for Rb+ and Cs+ transport. At an external Ca2+ concentration in the micromolar range the rate of transport decreased in the order: Na+ greater than K+ = H+ greater than Cs+ greater than Li+. Na+ fluxes depended in a sigmoidal way on the external Na+ concentration, suggesting that sodium ions move in pairs. The concentration dependence of uniport Na+ transport and that of Na+-stimulated Ca2+ efflux (exchange or antiport transport) were very similar.