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Am J Physiol Renal Physiol (January 6, 2004). doi:10.1152/ajprenal.00277.2003
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Submitted on August 6, 2003
Accepted on December 31, 2003

Increased expression and apical targeting of renal ENaC subunits in puromycin aminonucleoside-induced nephrotic syndrome in rats

Soo Wan Kim1, Weidong Wang1, Jakob Nielsen1, Jeppe Praetorius1, Tae-Hwan Kwon2, Mark A. Knepper3, Jorgen Frokiaer1, and Soren Nielsen1*

1 The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark
2 Department of Physiology, Dongguk University School of Medicine, Kyungju, Korea, Republic of
3 Laboratory and Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

* To whom correspondence should be addressed. E-mail: sn{at}ana.au.dk.

Nephrotic syndrome is often accompanied by sodium retention and generalized edema. However, the molecular basis for the decreased renal sodium excretion remains undefined. We hypothesized that ENaC subunit dysregulation may be responsible for the increased sodium retention. An experimental group of rats was treated with puromycin aminonucleoside (PAN, 180 mg/kg, i.v.), while the control group received only vehicle. After 7 days, PAN treatment induced significant proteinuria, hypoalbuminemia, decreased urinary sodium excretion and extensive ascites. The protein abundance of {alpha}ENaC and {beta}ENaC was increased in the inner stripe of the outer medulla (ISOM) and in the inner medulla (IM), but was not altered in cortex. {gamma}ENaC abundance was increased in cortex, ISOM and IM. Immunoperoxidase brightfield- and laser scanning confocal fluorescence-microscopy demonstrated increased targeting of {alpha}ENaC, {beta}ENaC and {gamma}ENaC subunits to the apical plasma membrane in the distal convoluted tubule (DCT2), connecting tubule, and cortical and medullary collecting duct segments. Immunoelectron microscopy further revealed an increased labeling of {alpha}ENaC in the apical plasma membrane of cortical collecting duct principal cells of PAN-treated rats, indicating enhanced apical targeting of {alpha}ENaC subunits. In contrast, the protein abundances of NHE3, BSC-1 and TSC were decreased. Moreover, the abundance of the {alpha}-1 subunit of the Na,K-ATPase was decreased in the cortex and ISOM, but remained unchanged in the IM. In conclusion, the increased or sustained expression of ENaC subunits combined with increased apical targeting in the DCT2, connecting tubule and collecting duct is likely to play a role in the sodium retention associated with PAN-induced nephrotic syndrome. The decreased abundance of NHE3, BSC-1, TSC and Na,K-ATPase may play a compensatory role to promote sodium excretion.




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