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Am J Physiol Regul Integr Comp Physiol 290: R479-R492, 2006. First published September 22, 2005; doi:10.1152/ajpregu.00512.2005
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WATER AND ELECTROLYTE HOMEOSTASIS

Ultrastructural localization of UT-A and UT-B in rat kidneys with different hydration status

Sun-Woo Lim,1,3 Ki-Hwan Han,4 Ju-Young Jung,1,3 Wan-Young Kim,1,3 Chul-Woo Yang,2,3 Jeff M. Sands,5 Mark A. Knepper,6 Kirsten M. Madsen,7 and Jin Kim1,3

1Department of Anatomy, 2Internal Medicine, and 3MRC for Cell Death Disease Research Center, The Catholic University of Korea, Seoul, Korea; 4Department of Anatomy, College of Medicine, Ewha Womans University, Seoul, Korea; 5Renal Division, Department of Medicine, Emory University School of Medicine, Atlanta, Georgia; 6Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland; and 7 Department of Medicine, University of Florida College of Medicine, Gainesville, Florida

Submitted 13 July 2005 ; accepted in final form 20 September 2005

Urea transport in the kidney is mediated by a family of transporter proteins, including renal urea transporters (UT-A) and erythrocyte urea transporters (UT-B). We aimed to determine whether hydration status affects the subcellular distribution of urea transporters. Male Sprague-Dawley rats were divided into three groups: dehydrated rats (WD) given minimum water, hydrated rats (WL) given 3% sucrose in water for 3 days before death, and control rats given free access to water. We labeled kidney sections with antibodies against UT-A1 and UT-A2 (L194), UT-A3 (Q2), and UT-B using preembedding immunoperoxidase and immunogold methods. In control animals, UT-A1 and UT-A3 immunoreactivities were observed throughout the cytoplasm in inner medullary collecting duct (IMCD) cells, and weak labeling was observed on the basolateral plasma membrane. UT-A2 immunoreactivity in the descending thin limbs (DTL) was observed mainly on the apical and basolateral membranes of type I epithelium, and very faint labeling was observed in the long-loop DTL at the border between the outer and inner medulla. UT-A1 immunoreactivity intensity was markedly lower, and UT-A3 immunoreactivity was higher in IMCD of WD vs. controls. UT-A2 immunoreactivity intensities in the plasma membrane and cytoplasm of type I, II, and III epithelia of DTL were greater in WD vs. controls. In contrast, UT-A1 expression was greater and UT-A2 and UT-A3 expressions were lower in WL vs. controls. The subcellular distribution of UT-A in DTL or IMCD did not differ between control and experimental animals. UT-B was expressed in the plasma membrane of the descending vasa recta of both control and experimental animals. UT-B intensity was higher in WD and lower in WL vs. controls. These data indicate that changes in hydration status over 3 days affected urea transporter protein expression without changing its subcellular distribution.

urea transporter A; urea transporter B; hydration status; subcellular localization



Address for reprint requests and other correspondence: J. Kim, Dept. of Anatomy, College of Medicine, The Catholic Univ. of Korea, 505 Banpo-Dong, Socho-Gu, Seoul 137-701, Korea (e-mail: jinkim{at}catholic.ac.kr)




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