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Am J Physiol Lung Cell Mol Physiol (June 18, 2004). doi:10.1152/ajplung.00340.2003
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Submitted on September 23, 2003
Accepted on June 12, 2004

Cyclic AMP-stimulated Na+ transport in H441 distal lung epithelial cells: Role of protein kinase A, phosphatidyl inositol 3-kinase and sgk1

Christie P. Thomas1*, Jason R. Campbell2, Patrick J. Wright1, and Russell F. Husted2

1 Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA; Veterans Affairs Medical Center, Iowa City, IA, USA
2 Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA

* To whom correspondence should be addressed. E-mail: christie-thomas{at}uiowa.edu.

H441 cells, a bronchiolar epithelial cell line, develop a cyclic AMP-regulated benzamil-sensitive Na+ transport pathway on permeable supports (AJP 282: L631-L641, 2002). To understand the molecular basis for the stimulation of Na+ transport, we delineated the role of specific intracellular pathways and examined the effect of cAMP on {alpha}, {beta}, {gamma}ENaC and sgk1 expression. Na+ transport increases within 5 minutes of cAMP stimulation and is sustained for over 24 hr. The sustained effect of cAMP on Na+ transport is abolished by LY294002, an inhibitor of PI3-Kinase, by H89, an inhibitor of PKA, or by SB202190, an inhibitor of p38 MAP kinase. The sustained effect of cAMP was associated with increases in {alpha}ENaC mRNA and protein but without a detectable increase in {beta}, {gamma}ENaC and sgk1. The early effect of cAMP on Na+ transport is brefeldin-sensitive and is mediated via protein kinase A. These results are consistent with a model where the early effect of cAMP is to increase trafficking of Na+ channels to the apical cell surface while the sustained effect requires the synthesis of {alpha}ENaC.




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