Vasopressin increases Na-K-2Cl cotransporter expression in thick ascending limb of Henle’s loop

Author:

Kim Gheun-Ho1,Ecelbarger Carolyn A.1,Mitchell Carter1,Packer Randall K.2,Wade James B.13,Knepper Mark A.1

Affiliation:

1. Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892;

2. Department of Biological Sciences, George Washington University, Washington, District of Columbia 20052; and

3. Department of Physiology, University of Maryland College of Medicine, Baltimore, Maryland 21201

Abstract

To investigate whether the enhancement of thick ascending limb (TAL) NaCl transport in response to long-term increases in circulating vasopressin concentration is associated with increased expression levels of the apical Na-K-2Cl cotransporter in the rat TAL, we have carried out immunoblotting and immunofluorescence studies using affinity-purified, peptide-directed antibodies. Semiquantitative immunoblotting studies demonstrated a marked increase (193% of controls) in Na-K-2Cl cotransporter band density in response to restriction of water intake to 15 ml/day for 7 days. In contrast, the expression levels of two other apical proteins of the TAL (the type 3 Na/H exchanger and Tamm-Horsfall protein) were unchanged in the outer medulla. A 7-day subcutaneous infusion of the V2receptor-selective vasopressin analog, 1-desamino-[8-d-arginine]vasopressin (DDAVP), to Brattleboro rats also markedly increased Na-K-2Cl cotransporter expression in the outer medulla (183% of controls). Immunofluorescence localization in outer medullary tissue sections confirmed the increase in Na-K-2Cl cotransporter expression in response to DDAVP. We conclude that vasopressin strongly upregulates the expression of the Na-K-2Cl cotransporter of the TAL and that it is likely to play an important role in the long-term regulation of the countercurrent multiplication system.

Publisher

American Physiological Society

Subject

Physiology

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