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Articles in PresS, published online ahead of print October 5, 2001
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00085.2001
Submitted on March 7, 2001
Accepted on October 1, 2001
1 Internal Medicine, University of Iowa, Iowa City, IA, USA
2 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, Bethesda, MD, USA
3 Internal Medicine, University of Iowa, Iowa City, IA, USA; Internal Medicine, Veterans Affairs Medical Center, Iowa City, IA, USA
* To whom correspondence should be addressed. E-mail: christie-thomas{at}uiowa.edu.
When grown on permeable supports, H441, a human bronchiolar epithelial cell line develops an amiloride-sensitive Na+ transport pathway that is glucocorticoid-regulated (JBC 274: 12431-12437). To understand the molecular basis for the electrogenic Na+ transport, we examined the effect of dexamethasone on 
,ß and 
ENaC and sgk1 mRNA expression and determined the biophysical properties of Na+ channels in these lung epithelial cells. Dexamethasone stimulated the expression of ,ß ,ENaC and sgk1 mRNA with the first effect seen by 1 hr. These effects were abolished by actinomycin D, but not by cycloheximide, indicating a direct stimulatory effect on ENaC and sgk1 mRNA synthesis. The glucocorticoid effect on transcription of ENaC mRNA was accompanied by a significant increase in ENaC protein levels. Glucocorticoids also stimulated ,ß and ENaC and sgk1 mRNA expression in A549 cells, a human alveolar type II cell line. To determine the biophysical properties of the Na+ channel, single channel currents were recorded from cell-attached H441 membrane patches. A Na+ selective channel with slow kinetics and a slope conductance of 10.8 pS was noted, properties similar to ,ß, ENaC expressed in X. Laevis oocytes. These experiments indicate that amiloride-sensitive Na+ transport is probably mediated through classic ENaC channels in human lung epithelia and that glucocorticoid-regulated Na+ transport is accompanied by increased transcription of each of the component subunits and sgk1.
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