Abstract
The cAMP-dependent activation of Cl− channels was studied in a bronchial epithelial cell line (16HBE14o-) in fast and slow whole-cell, and cell-attached patch-clamp experiments. The cells are known to express high levels of cystic fibrosis transmembrane conductance regulator mRNA and protein. Isoproterenol, forskolin and histamine (all 10 μmol/l) reversibly and significantly depolarized the membrane voltage (V m) and increased the whole-cell Cl− conductance significantly by 34.0±0.9 (n=3), 18.1±2.7 (n=50), and 25±4.5 (n=37) nS respectively. The effect of histamine was blocked by cimetidine (10 μmol, n=5) but not by diphenhydramine (10 μmol/l, n=4), which suggests binding of histamine to H2 receptors. The forskolin-induced current was not inhibited significantly by 4,4′-diisothiocyanatostilbene-2,2′-disulphonic acid (0.5 mmol/l, n=9) nor glibenclamide (10 μmol/l, n=3) and had an anion-permeability sequence of Cl−= Br−>I− (n=9). In cell-attached recordings forskolin (10 μmol/l) increased the conductance of the patched membrane from 65.5±13.6 pS to 150.8±33.2 pS (n=30). Although the conductance was increased significantly, clear ion-channel events occurring in parallel with the current activation were not detected in the cell attached membrane. In 4 out of 30 cell-attached recordings single-channel currents were observed. These channels, with a single-channel conductance of about 6 pS, were already active before forskolin was added. No effect of forskolin on the channel amplitude, open probability or kinetics of these channels was observed. From these data we conclude that the cAMP-induced conductance increase in 16HBE14o-cells can be correlated with the activation of very small and not resolvable (probably less than 2 pS) Cl− channels rather than with the activation of channels with a conductance of 6–10 pS.
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Kunzelmann, K., Koslowsky, T., Hug, T. et al. CAMP-dependent activation of ion conductances in bronchial epithelial cells. Pflugers Arch. 428, 590–596 (1994). https://doi.org/10.1007/BF00374582
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DOI: https://doi.org/10.1007/BF00374582