Two-pore-domain potassium channels support anion secretion from human airway Calu-3 epithelial cells.
Potassium channels are required for the absorption and secretion of fluids and electrolytes in epithelia. Calu-3 cells possess a secretory phenotype, and are a model human airway submucosal gland serous cell. Short-circuit current (I(sc)) recordings from Calu-3 cells indicated that basal anion secretion was reduced by apical application of the K+ channel inhibitors bupivicaine, lidocaine, clofilium, and quinidine. Application of riluzole resulted in a large increase in I(sc), inhibited by apical application of either bupivicane or the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel blocker DPC. These results suggested that one or more members of the two-pore-domain K+ (K(2P)) channel family could influence anion secretion. Using RT-PCR, we found that Calu-3 cells express mRNA transcripts for TASK-2 (KCNK5), TWIK-1 (KCNK1), TWIK-2 (KCNK6) and TREK-1 (KCNK2). TASK-2, TWIK-2 and TREK-1 protein were detected by Western blotting, while immunolocalization of polarized cells confirmed protein expression of TREK-1 and TWIK-2 at the plasma cell membrane. TASK-2 protein staining was localized to intracellular vesicles, located beneath the apical membrane. While the pro-secretory role of basolateral K+ channels is well established, we suggest that apically located K2P channels, not previously described in airway epithelial cells, also play an important role in controlling the rate of transepithelial anion secretion.
Davis
Kellie A
KA
Department of Physiology and Biophysics, Dalhousie University, Halifax, NS, B3H 4H7, Canada.
Cowley
Elizabeth A
EA
D000838:Anions / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D004847:Epithelial Cells / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D024683:Potassium Channels, Tandem Pore Domain / Q000502:physiology*; D020545:Respiratory Mucosa / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000838:Anions; D024683:Potassium Channels, Tandem Pore Domain