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Articles in PresS, published online ahead of print April 23, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00016.2002
Submitted on January 11, 2002
Accepted on April 17, 2002
1 National Heart, Lung, and Blood Institute, NIH, Bethesda, MD, USA
2 National Heart, Lung, and Blood Institute, NIH, Bethesda, MD, USA; Water and Salt Institute, University of Aarhus, Aarhus, Denmark
3 Water and Salt Institute, University of Aarhus, Aarhus, Denmark
4 Department of Internal Medicine, University of Iowa, Iowa City, Iowa, USA
5 Internal Medicine, University of Iowa, Iowa City, Iowa, USA
* To whom correspondence should be addressed. E-mail: knep{at}helix.nih.gov.
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 following initiation of dietary NaCl restriction. After equilibration on a NaCl-replete diet (2.0 meq/200gBW/day), rats were switched to a NaCl-deficient diet (0.02 meq/200gBW/day). Na excretion fell to 25% of baseline levels on day 1 followed by a further decrease to less than 4% of baseline levels 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 one-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. Ribonuclease 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.
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