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Increased proximal tubule Na,K‐ATPase and phosphorylation at Ser‐11 in 2‐kidney, 1‐clip Goldblatt hypertensive rat
Author(s) -
Yingst Douglas R.,
Kava Lauren,
Rossi Noreen F.,
Chow Albert,
Mattingly Raymond
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.605.14
Subject(s) - phosphorylation , endocrinology , medicine , chemistry , kidney , phosphatase , actin , angiotensin ii , renin–angiotensin system , blood pressure , biochemistry
Here we test the hypothesis that in the 2‐kidney, 1‐clip rat, in which hypertension develops due to increased angiotensin II levels, there is an increase in expression of Na,K‐ATPase and increased phosphorylation at sites that are known to regulate its activity (Ser‐11 and Ser‐18). Five week old Sprague Dawley rats underwent surgical placement of a 0.2 mm silver clip around the right renal artery or sham clipping. Six weeks later, blood pressure was measured by telemetry. Rats were then anesthetized, the kidney cortex harvested and minced, and Na,K‐ATPase from left and right kidneys extracted with RIPA buffer supplemented with 2% SDS, protease and phosphatase inhibitors. The amount of Na,K‐ATPase, the level of phosphorylation at Ser‐11 and Ser‐18, and the expression of β actin were measured by quantitative immunoblotting. Clipping increased the mean arterial pressure from 109 ± 7 to 147 ± 30 mm Hg (p= 0.001), increased the amount of α‐subunit of Na,K‐ATPase per mg protein by 24% (p=0.000), and increased phosphorylation of Ser‐11 per α‐subunit 29% (p=0.025). Clipping did not affect phosphorylation at Ser‐18 per α‐subunit (p=0.07) or expression of β actin (p=0.29). We conclude that there is increased expression of Na,K‐ATPase and phosphorylation of Ser‐11 in the kidney cortex during the development of hypertension in this model. Supported by 1R01‐HL079102 to N.F.R and R01‐DK‐60752 to D.Y.