Arginine Vasopressin regulates the renal Na-Cl and Na-K-Cl Cotransporters through With-No-Lysine Kinase 4 and Inhibitor 1 Phosphorylation

Am J Physiol Renal Physiol. 2023 Dec 14. doi: 10.1152/ajprenal.00343.2023. Online ahead of print.ABSTRACTVasopressin regulates water homeostasis via the V2 receptor in the kidney at least in part through protein kinase A (PKA) activation. Vasopressin, through an unknown pathway, upregulates the activity and phosphorylation of the Na+-Cl- cotransporter (NCC) and Na+-K+-2Cl- cotransporter 2 (NKCC2) by Ste20-related Proline/Alanine rich Kinase (SPAK) and Oxidative Stress Responsive kinase 1 (OSR1), which are regulated by the With No Lysine (K) kinase (WNK) family. Phosphorylation of WNK4 at PKA consensus motifs may be involved. Inhibitor 1 (I1), a Protein Phosphatase 1 (PP1) inhibitor, may also play a role. In HEK293 cells, we assessed the phosphorylation of WNK4, SPAK, NCC, or NKCC2 in response to forskolin or desmopressin. WNK4 and cotransporter phosphorylation was studied in desmopressin-infused WNK4-/- mice and in tubule suspensions. In HEK293 cells, only wild-type WNK4, but not WNK1, WNK3, or a WNK4 mutant lacking PKA phosphorylation motifs could upregulate SPAK or cotransporter phosphorylation in response to forskolin or desmopressin. I1 transfection maximized SPAK phosphorylation in response to forskolin in the presence of WNK4, but not of mutant WNK4 lacking PP1 regulation. We observed direct PP1 regulation of NKCC2 dephosphorylation, but not of NCC or SPAK in the absence of WNK4. WNK4-/- mice with desmopressin treatment did not increase SPAK/OSR1, NCC, or NKCC2 phosphor...
Source: Am J Physiol Renal P... - Category: Urology & Nephrology Authors: Source Type: research