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Department of Physiology, Dartmouth Medical School, Lebanon, New Hampshire 03756-0001
To study the role
of serum and glucocorticoid-inducible kinase-1 (SGK1) in mammalian
cells, we compared Na+ transport rates in wild-type (WT) M1
cortical collecting duct cells with M1 populations stably expressing
human full-length SGK1, NH2-terminal truncated (
N-60)
SGK1, "kinase-dead" (K127M) SGK1, and cells that have downregulated
levels of SGK1 mRNA (antisense SGK1). Basal rates of transepithelial
Na+ transport were highest in full-length SGK1 populations,
compared among the above populations. Dexamethasone treatment increased Na+ transport in WT and full-length SGK1 cells 2.7- and
2-fold, respectively. Modest stimulation of Na+ absorption
was detected after dexamethasone treatment in N-60 SGK1 populations.
However, N-60 SGK1 transport rates remained substantially lower than
WT values. Importantly, a combination of high insulin, dexamethasone,
and serum failed to significantly stimulate Na+ transport
in antisense or K127M SGK1 cells. Additionally, expression of antisense
SGK1 significantly decreased transepithelial resistance values.
Overall, we concluded that SGK1 is a critical component in
corticosteroid-regulated Na+ transport in mammalian
cortical collecting duct cells. Furthermore, our data suggest that the
NH2 terminus of SGK1 may contain a Phox homology-like
domain that may be necessary for effective Na+ transport.
aldosterone; epithelial sodium channel; M1 cell line; serum and glucocorticoid-inducible kinase 1; cortical collecting duct
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