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1 Physiology and Biophysics, USC Keck School of Medicine, Los Angeles, California, United States
2 Cell Biology, University of Aarhus Water and Salt Research Center, Aarhus, Denmark
* To whom correspondence should be addressed. E-mail: mcdonoug{at}usc.edu.
The distal convoluted tubule (DCT) apical Na+-Cl- cotransporter (NCC) is responsible for the reabsorption of 5 - 10% of filtered NaCl and is the target for thiazide diuretics. NCC abundance is increased during dietary NaCl restriction and by aldosterone, decreased during high salt diet, and mineralocorticoid blockade. This study tested the hypothesis that subcellular distribution of NCC is also regulated in response to changes in dietary salt. Six week old Sprague Dawley rats were fed a normal salt diet (NS = 0.4% NaCl) for 3 weeks, then switched to high salt diet (HS = 4% NaCl) for 3 weeks or low salt diet (LS = 0.07% NaCl) for 1 week. Under anesthesia, kidneys were excised, renal cortex dissected and NCC analyzed with specific antibodies after either: 1) density gradient centrifugation followed by immunoblot or 2) fixation followed by immunoelectron microscopy. HS diet decreased NCC abundance to 0.50 ±0.10 of levels in LS diet (1.00±0.23). HS diet also caused a redistribution of NCC from low to higher density membranes. Immunoelectron microscopy revealed that NCC resides predominantly in the apical membrane in rats fed LS diet and increases in sub-apical vesicles in rats fed HS diet. In conclusion, high salt diet provokes a rapid and persistent redistribution of NCC from apical to sub-apical membranes, a mechanism that would facilitate a homeostatic decrease in NaCl reabsorption in the DCT to compensate for increased dietary salt.
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