Membrane potential measurement in isolated outer hair cells of the guinea pig cochlea using conventional microelectrodes

Abstract

Membrane potential of the isolated outer hair cells (OHCs) from the guinea pig cochlea was measured using conventional microelectrodes filled with 200 mM KCl. The resting membrane potential during superfusion with the standard physiological saline solution containing 3.5 mM K + was −47.3 ± 1.4 mV ( N = 72), which was higher than those previously reported for isolated OHCs studied by using microelectrodes. Addition of ouabain (10 −5–10 −3 M), the specific Na +, K + ATPase inhibitor, depolarized the cell slowly and progressively, indicating the presence of low but definite Na +, K + ATPase activity in the plasma membrane of OHCs. The magnitude of membrane potential was mainly dependent on the extracellular K + concentration ([K +] o). A ten-fold increase of [K +] o depolarized the membrane potential by 49.6 ± 1.0 mV ( N = 58). A decrease of [Na +] o to one tenth of the control hyperpolarized the membrane potential by about 2 mV. Decreasing extracellular Cl − from 131.3 mM to 27.5 mM did not cause a significant change in the membrane potential. Using the Goldman-Hodgkin-Katz equation, assuming a negligible contribution of Cl − to the membrane potential and total monovalent cat ion concentration of the cytosol similar to the extracellular fluid, we calculated the permeability ratio of K + versus Na + to 131 ± 19 and intracellular K + concentration to 33.3 ± 1.9 mM.