Stretch-activated nonselective cation, Cl− and K+ channels in apical membrane of epithelial cells of Reissner's membrane

Abstract

Ion channels on the apical membrane of epithelial cells (the surface facing the endolymph) of acutely isolated Reissner's membrane from guinea-pig cochlea were investigated by using patch-clamp technique in cell-attached and inside-out configurations. Three types of ion channel were identified: namely, a stretch-activated nonselective cation, a chloride and a potassium channel. When the pipette was filled with high-K+ endolymph-like solution, the most significant channel activity was nonselective cation channels (85/110, 77% patches). The current versus voltage relationship was linear with a unitary conductance of 22.1±0.4 pS and reversal potential (Vr) of 2.3±0.8 mV (n=18). The channel exhibited a lower conductance (14.0±0.6 pS, n=8) to Ca2+. The open probability was low (NPo≅0.1) in cell-attached configuration under +60 mV pipette potential and increased when the membrane was stretched with negative pressure. The channel was blocked by 10 μM extracellular Gd3+. The two other types of channels were a small voltage-sensitive Cl− channel (6.0±0.3 pS; 91/99, 92% patches) and a K+ channel (≅30 pS; 29/191, 15% patches). These channels might play roles in the regulation of cell volume, in balancing the hydrostatic pressure across Reissner's membrane and in maintaining the electrochemical composition of endolymph.