Short-term stimulation of the thiazide-sensitive [Na.sup.+]-[Cl.sup.-] cotransporter by vasopressin involves phosphorylation and membrane translocation

Vasopressin influences salt and water transport in renal epithelia. This is coordinated by the combined action of [V.sub.2] receptor-mediated effects along distinct nephron segments. Modulation of NaCl reabsorption by vasopressin has been established in the loop of Henle, but its role in the distal convoluted tubule (DCT), an effective site for fine regulation of urinary electrolyte composition and the target for thiazide diuretics, is largely unknown. The [Na.sup.+]-[Cl.sup.-] cotransporter (NCC) of DCT is activated by luminal trafficking and phosphorylation at conserved N[H.sub.2]-terminal residues. Here, we demonstrate the effects of short-term vasopressin administration (30 min) on NCC activation in Brattleboro rats with central diabetes insipidus (DI) using the [V.sub.2] receptor agonist desmopressin (dDAVP). The fraction of NCC abundance in the luminal plasma membrane was significantly increased upon dDAVP as shown by confocal microscopy, immunogold cytochemistry, and Western blot, suggesting increased apical trafficking of the transporter. Changes were paralleled by augmented phosphorylation of NCC as detected by antibodies against phospho-threonine and phospho-serine residues (2.5-fold increase at Thr53 and 1 A-fold increase at Ser71). dDAVP-induced phosphorylation of NCC, studied in tubular suspensions in the absence of systemic effects, was enhanced as well (1.7-fold increase at Ser71), which points to the direct mode of action of vasopressin in DCT. Changes were more pronounced in early (DCT1) than in late DCT as distinguished by the distribution of 1l[beta]-hydroxysteroid dehydrogenase 2 in DCT2. These results suggest that the vasopressin-[V.sub.2] receptor-NCC signaling cascade is a novel effector system to adjust transepithelial NaC1 reabsorption in DCT. antidiuretic hormone; distal convoluted tubule; phosphorylation; sodium-chloride cotransporter trafficking doi:10.1152/ajprenal.00476.2009