Hypotonic stress upregulates β- and γ-ENaC expression through suppression of ERK by inducing MKP-1

Niisato N, Ohta M, Eaton DC, Marunaka Y. Hypotonic stress upregulates beta- and gamma-ENaC expression through suppression of ERK by inducing MKP-1. Am J Physiol Renal Physiol 303: F240-F252, 2012. First published May 9, 2012; doi:10.1152/ajprenal.00198.2011.-We investigated a physiological role for ERK, a member of the MAPK family, in the hypotonic stimulation of epithelial Na+ channel (ENaC)-mediated Na+ reabsorption in renal epithelial A6 cells. We show that hypotonic stress causes a major dephosphorylation of ERK following a rapid transient phosphorylation. PD98059 (a MEK inhibitor) increases dephosphorylated ERK and enhances the hypotonic-stress-stimulated Na+ reabsorption. ERK dephosphorylation is mediated by MAPK phosphatase (MKP). Hypotonic stress activates p38, which in turn induces MKP-1 and to a lesser extent MKP-3 mRNA expression. Inhibition of p38 suppresses MKP-1 induction, preventing hypotonic stress from dephosphorylating ERK. Inhibition of MKP-1 and -3 by the inhibitor NSC95397 also suppresses the hypotonicity-induced dephosphorylation of ERK. NSC95397 reduces both beta- and gamma-ENaC mRNA expression and ENaC-mediated Na+ reabsorption stimulated by hypotonic stress. In contrast, pretreatment with PD98059 significantly enhances mRNA and protein expression of beta- and gamma-ENaC even under isotonic conditions. However, PD98059 only stimulates Na+ reabsorption in response to hypotonic stress, suggesting that ERK inactivation by itself (i.e., under isotonic conditions) is not sufficient to stimulate Na+ reabsorption, even though ERK inactivation enhances beta- and gamma-ENaC expression. Based on these results, we conclude that hypotonic stress stimulates Na+ reabsorption through at least two signaling pathways: 1) induction of MKP-1 that suppresses ERK activity and induces beta- and gamma-ENaC expression, and 2) promotion of translocation of the newly synthesized ENaC to the apical membrane.