title: Expression of the chloride channel CLC-K in human airway epithelial cells.
abstract: Airway submucosal gland function is severely disrupted in cystic fibrosis (CF), as a result of genetic mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), an apical membrane Cl(-) channel. To identify other Cl(-) channel types that could potentially substitute for lost CFTR function in these cells, we investigated the functional and molecular expression of Cl(-) channels in Calu-3 cells, a human cell line model of the submucosal gland serous cell. Whole cell patch clamp recording from these cells identified outwardly rectified, pH- and calcium-sensitive Cl(-) currents that resemble those previously ascribed to ClC-K type chloride channels. Using reverse transcription polymerase chain reaction, we identified expression of mRNA for ClC-2, ClC-3, ClC-4, ClC-5, ClC-6, ClC-7, ClC-Ka, and ClC-Kb, as well as the common ClC-K channel beta subunit barttin. Western blotting confirmed that Calu-3 cells express both ClC-K and barttin protein. Thus, Calu-3 cells express multiple members of the ClC family of Cl(-) channels that, if also expressed in native submucosal gland serous cells within the CF lung, could perhaps act to partially substitute lost CFTR function. Furthermore, this work represents the first evidence for functional ClC-K chloride channel expression within the lung.
authors.affiliation: Department of Physiology and Biophysics, Dalhousie University, Halifax, NS B3H 1X5, Canada.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Mummery
authors.forename: Jennifer
authors.initials: J
authors.lastname: Killey
authors.forename: Paul
authors.initials: P
authors.lastname: Linsdell
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D018118:Chloride Channels / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004594:Electrophysiology; D004847:Epithelial Cells / Q000378:metabolism*; D006801:Humans; D018408:Patch-Clamp Techniques; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018118:Chloride Channels; D016207:Cytokines; C034477:RNA primers; C121169:cardiotrophin-like cytokine; D012313:RNA