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1 The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark; Department of Physiology, School of Medicine, Dongguk University, Kyungju, Korea, Republic of
2 The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark; Institute of Anatomy, University of Aarhus, Aarhus, Denmark
3 Laboratory of Kidney and Electrolyte Metabolism, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA
4 The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark; Institute of Experimental Clinical Research, University of Aarhus, Aarhus, Denmark
* To whom correspondence should be addressed. E-mail: sn{at}ana.au.dk.
Angiotensin II (AngII) is involved in the reabsorption of Na+ and HCO3- in the proximal tubule. The kidney thick ascending limbs (TAL) are also involved in the Na+ and HCO3- reabsorption. In the present study we examined the effect of AngII treatment of 50ng/min/Kg (AngII-50), 100ng/min/Kg (AngII-100), 200ng/min/Kg (AngII-200), s.c., for 7d on the abundance of TAL Na transporters NHE3, BSC-1, 
1-subunit of Na,K-ATPase and electroneutral NBCn1. All rats received a fixed daily food and water intake by ration feeding. Semiquantitative immunoblotting revealed that NHE3 abundance in the ISOM of AngII-treated rats was significantly increased: 179±28% (AngII-50, n=5), 166±23% (AngII-100, n=7) and 167±19% (AngII-200, n=4) of control levels (n=6, P<0.05). In contrast, treatment with lower doses of AngII was not associated with changes in NHE3 abundance in ISOM: 107±25% (AngII-12.5, n=5) and 123±18% (AngII-25, n=5). The abundance of BSC-1 in the ISOM was also increased to 187±28% (AngII-50), 162±23% (AngII-100) and 166±19% (AngII-200) of control levels (P<0.05), but there were no changes in the abundance of Na,K-ATPase and NBCn1. Immunocytochemistry confirmed the increase in NHE3 and BSC-1 labeling in mTAL. In contrast, in the renal cortex and OSOM, NHE3 abundance was unchanged, while immunocytochemistry and revealed an increased NHE3 labeling of the proximal tubule brush border suggesting subcellular redistribution of NHE3 or differential protein-protein interaction in the absence of changes in total proximal tubule abundance. Despite this, AngII-treated rats (50ng/min/Kg for 5d, n=6) had higher urine pH as compared with controls. NH4Cl-loading completely blocked all effects of AngII infusion on NHE3 and BSC-1, pointing to a potential role of pH as a mediator of these effects. In conclusion, increased abundance of NHE3 and BSC-1 in mTAL cells as well as increased NHE3 in the proximal tubule brush border may contribute to an enhanced renal Na+ and HCO3- reabsorption in response to Ang II.
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