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1 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-1603; 2 The Water and Salt Institute, University of Aarhus, DK-8000 Aarhus C, Denmark; and 3 Department of Internal Medicine, University of Iowa, and Veterans Affairs Medical Center, Iowa City, Iowa 52242
We have used
peptide-directed antibodies to each major renal Na transporter and
channel proteins to screen renal homogenates for changes in Na
transporter protein expression after initiation of dietary NaCl
restriction. After equilibration on a NaCl-replete diet (2.0 meq · 200 g body
wt
1 · day1), rats were switched to
a NaCl-deficient diet (0.02 meq · 200 g body
wt1 · day1). Na excretion fell to
25% of baseline levels on day 1, followed by a further
decrease <4% of baseline levels on day 3, of NaCl restriction. The decreased Na excretion at day 1 occurred
despite the absence of a significant increase in plasma aldosterone
level or in the abundance of any of the major renal Na transporters. However, after a 1-day lag, plasma aldosterone levels increased in
association with increases in abundances of three aldosterone-regulated Na transporter proteins: the thiazide-sensitive Na-Cl cotransporter (NCC), the 
-subunit of the amiloride-sensitive epithelial Na channel
(-ENaC), and the 70-kDa form of 
-ENaC. RNase protection assays of
transporter mRNA levels revealed an increase in renal -ENaC mRNA
coincident with the increase in -ENaC protein abundance. However,
there was no change in NCC mRNA abundance, suggesting that the increase
in NCC protein in response to dietary NaCl restriction was not a result
of altered gene transcription. These results point to early regulatory
processes that decrease renal Na excretion without an increase in the
abundance of any Na transporter, followed by a late
aldosterone-dependent response associated with upregulation of NCC and ENaC.
aldosterone; distal convoluted tubule; collecting duct; type 3 sodium/hydrogen exchanger; bumetanide-sensitive type 2 sodium-potassium-2 chloride cotransporter; thiazide-sensitive sodium-chloride cotransporter; epithelial sodium channel
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