A synthetic peptide based on a glycine-gated chloride channel induces a novel chloride conductance in isolated epithelial cells.

CK(4)-M2GlyR, an aqueous soluble peptide derived from the transmembrane M2 segment of the glycine-gated Cl(-) channel found in postsynaptic membranes of the central nervous system, has previously been shown to increase transepithelial Cl(-) and fluid secretion of epithelial monolayers. The goal of this study was to determine whether CK(4)-M2GlyR exerts these effects via formation of a novel chloride conductance pathway, modulation of endogenous chloride channel activity, or a combination of these effects. Ionic currents were recorded from isolated epithelial cells before and after treatment with the peptide using the whole-cell configuration of the patch-clamp technique. CK(4)-M2GlyR increased whole-cell Cl(-) currents in all epithelial cell lines that were studied, including: Madin-Darby canine kidney cells, a human colonic epithelial cell line (T84), and airway epithelial cells derived from a human cystic fibrosis patient (IB3-1). No evidence was found for modulation of endogenous Cl(-) channels by CK(4)-M2GlyR based on both the electrophysiological properties of the observed currents and the pharmacological profile of the CK(4)-M2GlyR-induced current. These results suggest that CK(4)-M2GlyR increases Cl(-) permeability in epithelial cells directly, by forming a distinct conduction pathway in cell membranes.