title: Mechanisms of angiotensin II stimulation of NCC are time-dependent in mDCT15 cells.
abstract: Angiotensin II (ANG II) increases thiazide-sensitive sodium-chloride cotransporter (NCC) activity both acutely and chronically. ANG II has been implicated as a switch that turns WNK4 from an inhibitor of NCC into an activator of NCC, and ANG II's effect on NCC appears to require WNK4. Chronically, ANG II stimulation of NCC results in an increase in total and phosphorylated NCC, but the role of NCC phosphorylation in acute ANG II actions is unclear. Here, using a mammalian cell model with robust native NCC activity, we corroborate the role that ANG II plays in WNK4 regulation and clarify the role of Ste20-related proline alanine-rich kinase (SPAK)-induced NCC phosphorylation in ANG II action. ANG II was noted to have a biphasic effect on NCC, with a peak increase in NCC activity in the physiologic range of 10(-11) M ANG II. This effect was apparent as early as 15 min and remained sustained through 120 min. These changes correlated with significant increases in NCC surface protein expression. Knockdown of WNK4 expression sharply attenuated the effect of ANG II. SPAK knockdown did not affect ANG II action at early time points (15 and 30 min), but it did attenuate the response at 60 min. Correspondingly, NCC phosphorylation did not increase at 15 or 30 min, but increased significantly at 60 min. We therefore conclude that within minutes of an increase in ANG II, NCC is rapidly trafficked to the cell surface in a phosphorylation-independent but WNK4-dependent manner. Then, after 60 min, ANG II induces SPAK-dependent phosphorylation of NCC.
authors: Department of Medicine, University of Chicago, Chicago, Illinois; bko@uchicago.edu.
authors: Benjamin
authors: B
authors: Ko
authors: Division of Nephrology, Department of Medicine, Emory University, Atlanta, Georgia; and.
authors: Abinash
authors: A
authors: Mistry
authors: Department of Medicine, University of Chicago, Chicago, Illinois;
authors: Lauren
authors: L
authors: Hanson
authors: Division of Nephrology, Department of Medicine, Emory University, Atlanta, Georgia; and.
authors: Rickta
authors: R
authors: Mallick
authors: Division of Nephrology, Department of Medicine, Emory University, Atlanta, Georgia; and Atlanta Veteran's Administration Medical Center, Decatur, Georgia.
authors: Robert S
authors: RS
authors: Hoover
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D049992:Sodium Chloride Symporters / Q000378:metabolism*; D013997:Time Factors
chemical_list: D049992:Sodium Chloride Symporters; D000804:Angiotensin II
keywords: WNK4 expression; distal convoluted tubule; thiazide-sensitive sodium-chloride cotransporter
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