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• W2149017264 abstract "Intermediate-conductance K + (Kcnn4) channels in the apical and basolateral membranes of epithelial cells play important roles in agonist-induced fluid secretion in intestine and colon. Basolateral Kcnn4 channels have been well characterized in situ using patch-clamp methods, but the investigation of Kcnn4 channels in apical membranes in situ has been hampered by a layer of mucus that prevents seal formation. In the present study, we used patch-clamp methods to characterize Kcnn4 channels in the apical membrane of IEC-18 cells, a cell line derived from rat small intestine. A monolayer of IEC-18 cells grown on a permeable support is devoid of mucus, and tight junctions enable selective access to the apical membrane. In inside-out patches, Ca 2+ -dependent K + channels observed with iberiotoxin (a Kcnma1/large-conductance, Ca 2+ -activated K + channel blocker) and apamin (a Kcnn1–3/small-conductance, Ca 2+ -activated K + channel blocker) present in the pipette solution exhibited a single-channel conductance of 31 pS with inward rectification. The currents were reversibly blocked by TRAM-34 (a Kcnn4 blocker) with an IC 50 of 8.7 ± 2.0 μM. The channels were not observed when charybdotoxin, a peptide inhibitor of Kcnn4 channels, was added to the pipette solution. TRAM-34 was less potent in inhibiting Kcnn4 channels in patches from apical membranes than in patches from basolateral membranes, which was consistent with a preferential expression of Kcnn4c and Kcnn4b isoforms in apical and basolateral membranes, respectively. The expression of both isoforms in IEC-18 cells was confirmed by RT-PCR and Western blot analyses. This is the first characterization of Kcnn4 channels in the apical membrane of intestinal epithelial cells." @default.
• W2149017264 created "2016-06-24" @default.
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• W2149017264 date "2011-11-01" @default.
• W2149017264 modified "2023-09-24" @default.
• W2149017264 title "Characteristics of Kcnn4 channels in the apical membranes of an intestinal epithelial cell line" @default.
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• W2149017264 doi "https://doi.org/10.1152/ajpgi.00558.2010" @default.
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