The application of patch clamp methods to ocular epithelia

Abstract

The techniques of patch voltage clamping and whole cell clamping have been applied to the lenses and corneas of several species of animals. Numerous ion channels have been found in the basal and apical membranes of lens epithelial cells, anterior and posterior surface lens fibers, apical membrane of corneal endothelial cells, and apical membrane of the second layer of corneal epithelial cells. No ion channels have been found in deep lens fiber membranes to date. There are 9-11 different kinds of potassium channels in ocular epithelial membranes, several different kinds of non-selective cation channels, and one non-selective channel with a large unit conductance. Sodium selective channels are seen only rarely while chloride selective channels have not been seen at all. Several channels have not yet been identified unequivocally. Using the gigohm seal technique, it is possible to show that the frog lens epithelial cell membrane is dominated by potassium channels. Also, a technique is described for using the reversal potential of a 25-30 pS non-selective cation channel to measure the resting voltage of epithelial cells without penetrating them. The results of lens ion channel localization studies are in only qualitative agreement with previous lens channel localization studies which used whole lens impedance and ion substitution techniques. Limitations of using the patch clamp for ion channel localization are presented.