title: The chloride channel ClC-4 co-localizes with cystic fibrosis transmembrane conductance regulator and may mediate chloride flux across the apical membrane of intestinal epithelia.
abstract: Cystic fibrosis (CF) causing mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) lead to mislocalization of CFTR protein from the brush border membrane of epithelial tissues and/or its dysfunction as a chloride channel. In initial reports, it was proposed that certain channels from the ClC family of chloride channels may provide compensatory or alternative pathways for epithelial chloride secretion in tissues from cystic fibrosis patients. In the present work, we provide the first evidence that ClC-4 protein is functionally expressed on the surface of the intestinal epithelium and hence, is appropriately localized to act as a therapeutic target in this CF-affected tissue. We show using confocal and electron microscopy that ClC-4 co-localizes with CFTR in the brush border membrane of the epithelium lining intestinal crypts in mouse and human tissues. In Caco-2 cells, a cell line thought to model human enterocytes, ClC-4 protein is expressed on the cell surface and also partially co-localizes with EEA1 and transferrin, marker molecules of early and recycling endosomes, respectively. Hence, like CFTR, ClC-4 may cycle between the plasma membrane and endosomal compartment. Furthermore, we show that ClC-4 functions as a chloride channel on the surface of these epithelial cells as antisense ClC-4 cDNA expression reduced the amplitude of endogenous chloride currents by 50%. These studies provide the first evidence that ClC-4 is endogenously expressed and may be functional in the brush border membrane of enterocytes and hence should be considered as a candidate channel to provide an alternative pathway for chloride secretion in the gastrointestinal tract of CF patients.
authors.affiliation: Program in Structural Biology, Research Institute, Hospital for Sick Children, Toronto M5S 1G8, Canada.
authors.forename: Raha
authors.initials: R
authors.lastname: Mohammad-Panah
authors.forename: Cameron
authors.initials: C
authors.lastname: Ackerley
authors.forename: Johanna
authors.initials: J
authors.lastname: Rommens
authors.forename: Monideepa
authors.initials: M
authors.lastname: Choudhury
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Wang
authors.forename: Christine E
authors.initials: CE
authors.lastname: Bear
mesh_terms: D018938:Caco-2 Cells; D018118:Chloride Channels / Q000032:analysis / Q000502:physiology*; D002712:Chlorides / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000032:analysis / Q000502:physiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*
chemical_list: C505032:CFTR protein, human; D018118:Chloride Channels; D002712:Chlorides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator