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Articles in PresS, published online ahead of print November 20, 2001
Am J Physiol Renal Physiol, 10.1152/ajprenal.00298.2001
Submitted on September 21, 2001
Accepted on November 13, 2001
1 Physiology, University of Southern California Keck School of Medicine, LA, CA, USA
2 Pharmaceutical Scinece, University of Southern California School of Pharmacy, LA, CA, USA; Physiology, University of Southern California Keck School of Medicine, LA, CA, USA
* To whom correspondence should be addressed. E-mail: mcdonoug{at}hsc.usc.edu.
Acute hypertension rapidly decreases proximal tubule (PT) Na+ reabsorption, facilitated by a redistribution of PT Na+/H+ exchangers (NHE3) out of the apical brush border, increasing NaCl at the macula densa, the signal for autoregulation of renal blood flow and GFR. This study aimed to determine whether NHE3 activity per transporter decreases during acute hypertension and the time dependence of the response. Blood pressure was elevated by 50-60 mmHg in male Sprague-Dawley rats for 5 or 30 min by constricting arteries. Renal cortical membranes were fractionated by density gradient centrifugation. NHE3 transport activity was assayed as rate of appearance of acridine orange (AO) from AO loaded vesicles in response to an inwardly directed Na+ gradient. After 5 minutes hypertension 20% of total NHE3 protein, assayed by immunoblot, redistributed from low density apical membranes to mid density membranes enriched in intermicrovillar cleft markers; by 30 min a similar percentage shifted to heavier density membranes containing markers of endosomes. NHE3 activity shifted to higher density membranes along with NHE3 protein, that is, no change in activity/transporter during acute hypertension. Confocal analysis of NHE-3 distribution also verified removal from apical microvilli and appearance in subapical vesicles. We conclude that the decrease in renal PT Na+ transport during acute hypertension is mediated by removal of transport competent NHE3 from the apical brush border to sub-apical and internal reserves.
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