Electrodiffusion of Cl⁻ and K⁺ in epithelial membranes reconstituted into planar lipid bilayers

Abstract

An electrodiffusive permeability for Cl⁻, its activation by low extracellular Cl⁻-concentrations and the interaction between electrodiffusive fluxes of Cl⁻ and K⁺ are demonstrated in the ventricular membranes from the epithelium of the bovine choroid plexus. Membranes were fused into artificial lipid bilayers formed at the tip of micropipettes. What is thought to be the cytoplasmic side of the membrane (the trans-side or the inside of pipette) was clamped at negative potentials (0 to −90 mV). Under these conditions the current was discrete, fluctuating less than 2 pA. With Cl⁻ as the only conducting ion on the two sides we observed a small electrodiffusive permeability which was reduced by bumetanide or furosemide by 62%. When the outside solution was rendered Cl⁻-free then the permeability to Cl⁻ increased by a factor of 2–5; this activation was reduced by bumetanide or furosemide by about 80%. We observed an interaction between inwards movements of K⁺ and outwards movements of Cl⁻ via the activated permeability: The total current was smaller than the sum of the expected inward K⁺-current and the expected outward activated Cl⁻-current. Bumetanide or furosemide increased the total current; apparently the loss of current carried by Cl⁻ was smaller than the gain in current carried by K⁺. The presence of K⁺ on both sides of the membrane was a condition for this interaction.