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Am J Physiol Renal Physiol 284: F778-F787, 2003. First published December 27, 2002; doi:10.1152/ajprenal.00088.2002
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Vol. 284, Issue 4, F778-F787, April 2003

Cycloheximide increases glucocorticoid-stimulated alpha -ENaC mRNA in collecting duct cells by p38 MAPK-dependent pathway

Omar A. Itani1,2, Kristyn L. Cornish1, Kang Z. Liu1, and Christie P. Thomas1,2,3

1 Department of Internal Medicine, and 2 Graduate Program in Molecular Biology, University of Iowa College of Medicine, and 3 Veterans Affairs Medical Center, Iowa City, Iowa 52242

Aldosterone and glucocorticoids (GCs) stimulate Na+ reabsorption in the collecting ducts by increasing the activity of the epithelial Na+ channel (ENaC). Our laboratory has used Madin-Darby canine kidney-C7 cells to demonstrate that this effect is associated with an increase in alpha -ENaC gene transcription (Mick VE, Itani OA, Loftus RW, Husted RF, Schmidt TJ, and Thomas CP, Mol Endocrinol 15: 575-588, 2001). Cycloheximide (CHX) superinduced the GC-stimulated alpha -ENaC expression in a dose-dependent manner, but had no effect on basal or aldosterone-stimulated alpha -ENaC expression, whereas anisomycin inhibited basal and corticosteroid-stimulated alpha -ENaC expression. The superinduction of alpha -ENaC expression was also seen with hypotonicity, was blocked by RU-38486, and was independent of protein synthesis. CHX had no effect on alpha -ENaC mRNA half-life, confirming that its effect was via an increase in alpha -ENaC transcription. The effect of CHX and hypotonicity on alpha -ENaC expression was abolished by SB-202190, indicating an effect mediated via p38 MAPK. Consistent with this scheme, CHX increased pp38 and MKK6, an upstream activator of p38, stimulated alpha -ENaC promoter activity. These data confirm a model in which CHX activates p38 in Madin-Darby canine kidney-C7 cells to increase alpha -ENaC gene transcription in a GC-dependent manner.

epithelial sodium channel; aldosterone; glucocorticoid; gene regulation


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