2004 Global Researcher Conference Proceeding

April 09 - 11, 2004

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Conference: 2004 Global Researcher Conference
Title: Urinary Concentrating Defect in Mice With Selective Deletion of Collecting-Duct Urea Transporter Isoforms, UT-A1 and UT-A3
Authors: Fenton, Robert; Chou, Chung-Lin; Stewart, Gavin; Smith, Craig P.; Knepper, Mark
Institutions: National Heart, Lung and Blood Institute, National Institutes of Health, University of Manchester
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Fenton Specialized transporter proteins; the products of two closely related genes, UT-A (Slc14a2) and UT-B (Slc14a1), modulate the movement of urea across cell membranes. To further examine the role of UT-A urea transporters in renal function we have selectively disrupted expression of the two known inner medulla collecting duct (IMCD) isoforms of UT-A, namely UT-A1 and UT-A3, producing UT-A1/3-/- mice. Knockout mice showed no evidence of UT-A1 or UT-A3 protein in the IMCD by both immunoblotting and immunocytochemistry, whereas expression of UT-A2 in the thin descending limbs of Henle’s loop was unchanged. Isolated perfused tubule studies of IMCDs from UT-A1/3-/- mice demonstrated a complete absence of phloretin-sensitive or vasopressin-stimulated urea transport with urea permeabilities similar to those calculated due to simple lipid phase diffusion. Analysis of growth by mouse weight revealed no differences between the genotypes and UT-A deficient mice had normal behavior, appearance and activity. On a 20% protein diet, and in comparison to littermate wild-type controls, UT-A1/3-/- mice exhibited significantly greater fluid consumption and urine flow. Under these conditions, UT-A1/3-/- urine osmolality was also significantly lower than wild-type mice. After a 24 hr water restriction, UT-A1/3-/- mice exhibited no ability to decrease urine flow and could not significantly raise their maximal urinary osmolality above that observed under basal conditions. Furthermore, during this 24 hr water restriction UT-A1/3-/- mice became lethargic and their body weight decreased by 21± 0.3% compared to 4.3 ± 1.2% in wild-type mice. The reduced concentrating ability observed was markedly diminished when urea excretion was decreased by dietary protein restriction (4% by weight), consistent with the Berliner hypothesis (1958) stating that the chief role of IMCD urea transport in the urinary concentrating mechanism is the prevention of urea-induced osmotic diuresis.
UT-A and UT-B transporter proteins help urea cross cell membranes. Fenton, et al., examined the role UT-A plays in kidney function. Mice lacking UT-A1 and UT-A3 were developed. These mice could not transport urea across cell membranes. On the same diet as control mice, the experimental mice consumed significantly more water and urinated significantly more than the control group. The experimental mice’s urine was far more dilute than the control group’s. After a 24-hour water restriction, the experimental mice still produced large volumes of dilute urine. During the water restriction, they became lethargic and lost weight. When the protein content of the mice’s diet was reduced from 20% to 4% and urea production decreased, their ability to concentrate urine improved markedly.