Modulation of membrane conductance in rods of Bufo marinus by intracellular calcium ion

Abstract

Double-barrel micropipettes were used to pressure-inject EGTA into the outer segments of rods in the isolated retina of Bufo marinus. We used these pipettes to point voltage clamp the cell to its resting membrane voltage during the injection of EGTA in order to prevent changes in membrane voltage from occurring. The input conductance of the rod was assessed by measuring the incremental membrane current required to hyperpolarize the membrane by less than or equal to 10 mV. When the retina was bathed in normal Ringer solution, the injection of EGTA during point voltage clamp evoked an inward membrane current and in increase in input conductance. This observation is consistent with an EGTA-evoked increase in conductance for an ion with an equilibrium potential more depolarized than the resting membrane potential. Injections of control solutions that did not contain EGTA had no effect. The effects of injected EGTA were not altered by variations in the pH or buffering capacity of the injection solution, or by the addition of equimolar Mg2+. Furthermore, injections of a solution containing equimolar Ca2+ and EGTA were without effect. Thus, the observed effects of injected EGTA were due to the lowering of the [Ca2+]i. Replacement of extracellular Na+ with choline+ abolished both the response to light and the EGTA-evoked increase in input conductance. A low [Na+]o solution containing 10(-8) M-Ca2+ reduced the response to injected EGTA by approximately the same amount as it reduced the response to light. Replacement of extracellular Cl- by methanesulphonate was without significant effect on either the response to light or to injected EGTA. These results are consistent with the interpretation that a lowered [Ca2+]i increases primarily the sodium conductance, gNa, of the plasma membrane of the rod outer segment. The conductance that is affected by a lowered [Ca2+]i appears to have the same specificity as the light-dependent conductance. This conclusion is consistent with a hypothesis for visual transduction involving modulation of gNa by light-evoked changes in the [Ca2+]i.