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Departments of 1Anesthesiology and 2Physiology and Biophysics, Schools of Medicine and Dentistry, University of Alabama at Birmingham, Birmingham, Alabama 35294
Submitted 2 December 2002 ; accepted in final form 11 April 2003
We recorded apical membrane potentials (Va) of H441 cells [a human lung cell line exhibiting both epithelial Na+ (ENaC) and CFTR-type channels] grown as confluent monolayers, using the microelectrode technique in current-clamp mode before, during, and after perfusion of the apical membranes with 10 µM forskolin. When perfused with normal Ringer solution, the cells had a Va of -43 ± 10 mV (means ± SD; n = 31). Perfusion with forskolin resulted in sustained depolarization by 25.0 ± 3.5 mV (means ± SD; n = 23) and increased the number, open time, and the open probability of a 4.2-pS ENaC. In contrast to a previous report (Jiang J, Song C, Koller BH, Matthay MA, and Verkman AS. Am J Physiol Cell Physiol 275: C1610–C1620, 1998), no transient hyperpolarization was observed. The forskolin-induced depolarization of Va was almost totally prevented by pretreatment of monolayers with 10 µM amiloride or by substitution of Na+ ions in the bath solution with N-methyl-D-glucamine. These findings indicate that cAMP stimulation of Na+ influx across H441 confluent monolayers results from activation of an amiloride-sensitive apical Na+ conductance and not from Va hyperpolarization due to Cl- influx through CFTR-type channels.
forskolin; epithelial sodium channel; cystic fibrosis trans-membrane conductance regulator; current clamp; patch clamp; single channel currents; amiloride; glibenclamide
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