Multi-electrode system for measurement of transmembrane ion-fluxes through living epithelial cells

Abstract

Cystic Fibrosis (CF) is the most common fatal human genetic disease. It is caused by the defect in a single anion channel protein which affects ion and water transport across the epithelial tissue. A flat multi-electrode platform of diameter 12mm, allowing for measurement of four ions: sodium, potassium, hydrogen and chloride by exchangeable/replaceable ion-selective electrodes is described. The measurement is possible owing to the architecture of the platform which accommodates all the electrodes and inlets/outlets. The platform fits to the cup and operates in a small volume of the solution bathing the living epithelial cell layer (membrane) deposited on a porous support of the cup, which allows for effective monitoring of ion concentration changes. By applying two multi-electrode platforms, it is possible to measure the ion transmembrane fluxes. The inlet and outlet tubes in the platforms allow for on-fly change of the calibrants, ion-concentration changes and ion channel blockers. Using different ion-concentration gradients and blockers of ion-transporting molecules we show for the first time that sodium ions flow from the basolateral to apical face of the cell monolayer via a paracellular route and return also via a transcellular one, while chloride anions are transported back and forth exclusively via a transcellular route.