A mutation that increases a novel calcium-activated potassium conductance of Paramecium tetraurelia

Abstract

Under two-electrode voltage clamp, a mutant of P. tetraurelia, restless (rst/rst), showed a large increase in induced current and an outward tail current when compared to the wild-type cell for hyperpolarizing voltage steps. An increase in the induced and tail currents is also observed for depolarizing voltage steps. The larger current during voltage steps and tail in the mutant were eliminated by the use of CsCl-filled electrodes and tetraethylammonium ion (TEA+) in the bath solution, characterizing the lesion as affecting a K+ conductance. Ionophoretic injection of ethylene glycol bis-(beta-aminoethyl ether) n,n,n',n-tetraacetic acid (EGTA) to buffer internal Ca2+ concentration reduced the increased K+ current and tail of the restless cell, indicating Ca2+ activation of the K+ current. Time course and amplitude of remaining currents after blockage of K+ conductances with Cs+ and TEA+ were similar in wild-type and restless cells suggesting no restless defect in entry of calcium. The Ca2+-activated sodium current was similar in the mutant to that in wild type arguing against a defect in calcium regulation activating the K+ channel in the restless cell. We conclude that the restless mutation alters a Ca2+-activated potassium conductance other than the one previously described. The multiplicity of Ca2+-activated potassium conductances in Paramecium is discussed.