Calmodulin-dependent regulation of hypotonicity-induced translocation of ENaC in renal epithelial A6 cells.
Hypotonicity stimulates translocation of epithelial Na(+) channel (ENaC) to the apical membrane from the intracellular store site of ENaC by activating protein tyrosine kinase (PTK) in renal epithelial A6 cells. Based upon the fact that calmodulin shows its action on other enzymes through PTK caused phosphorylation of tyrosine residues of calmodulin itself, we studied whether a calmodulin-dependent pathway is involved in the action of hypotonicity on ENaC. W7, an antagonist of calmodulin, diminished the stimulatory action of hypotonicity on ENaC, irrespective of W7 treatment before or after application of hypotonicity. Calmodulin is known to regulate three pathways: (1) protein phosphatase 2B (PP2B), (2) Ca(2+)/calmodulin-dependent protein kinase II (CaMK II), and (3) myosin light chain kinase (MLCK). Pretreatment with cyclosporin A, an inhibitor of PP2B, did not influence the hypotonicity action on ENaC. The hypotonicity action on ENaC was partially inhibited by pretreatment with KN93, an inhibitor of CaMK II, but not by addition of KN93 after hypotonic stimulation had been applied. ML-7, an inhibitor of MLCK, showed the action similar to KN93. These observations indicate that: (1) the hypotonicity-induced translocation of ENaC depends on CaMK II and MLCK and (2) ENaC translocated to the apical membrane by hypotonicity is maintained in its activity and/or stability at the apical membrane through a calmodulin-dependent pathway.
Tokuda
Shinsaku
S
Department of Cellular and Molecular Physiology, Kyoto Prefectural University of Medicine, Kyoto 602-084, Japan.
Niisato
Naomi
N
Morisaki
Shinsuke
S
Marunaka
Yoshinori
Y
D002147:Calmodulin / Q000502:physiology*; D002460:Cell Line; D016572:Cyclosporine / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D053503:Epithelial Sodium Channels; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D009994:Osmolar Concentration; D021381:Protein Transport; D015222:Sodium Channels / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology
D002147:Calmodulin; D053503:Epithelial Sodium Channels; D015222:Sodium Channels; D013449:Sulfonamides; C017967:W 7; D016572:Cyclosporine