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Am J Physiol Renal Physiol 291: F1033-F1044, 2006. First published June 6, 2006; doi:10.1152/ajprenal.00086.2006
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Locations, abundances, and possible functions of FXYD ion transport regulators in rat renal medulla

Kaarina Pihakaski-Maunsbach,1,2 Henrik Vorum,3 Bent Honoré,3 Shigeki Tokonabe,1,2 Jørgen Frøkiær,1,4 Haim Garty,5 Steven J. D. Karlish,5 and Arvid B. Maunsbach1,2

1The Water and Salt Research Center, 2Department of Cell Biology, Institute of Anatomy, 3Institute of Medical Biochemistry, and 4Department of Clinical Physiology, University of Aarhus, Aarhus, Denmark; and 5Department of Biological Chemistry, Weizmann Institute of Science, Rehovoth, Israel

Submitted 14 March 2006 ; accepted in final form 31 May 2006

The {gamma}-subunit of Na-K-ATPase (FXYD2) and corticosteroid hormone-induced factor (CHIF; FXYD4) are considered pump regulators in kidney tubules. The aim of this study was to expand the information about their locations in the kidney medulla and to evaluate their importance for electrolyte excretion in an animal model. The cellular and subcellular locations and abundances of {gamma} and CHIF in the medulla of control and sodium-depleted rats were analyzed by immunofluorescence and immunoelectron microscopy and semiquantitative Western blotting. The results showed that antibodies against the {gamma}-subunit COOH terminus and splice variant {gamma}a, but not splice variant {gamma}b, labeled intercalated cells, but not principal cells, in the initial part of the inner medullary collecting duct (IMCD1). In subsequent segments (IMCD2 and IMCD3), all principal cells exhibited distinct basolateral labeling for both the {gamma}-subunit COOH terminus, splice variant {gamma}a, and CHIF. Splice variant {gamma}b was abundant in the inner stripe of the outer medulla but absent in the inner medulla (IM). Double labeling by high-resolution immunoelectron microscopy showed close structural association between CHIF and the Na-K-ATPase {alpha}1-subunit in basolateral membranes. The present observations provide new information about the cellular and subcellular locations of {gamma} and CHIF in the renal medulla and show a new {gamma} variant in the IM. Extensive NaCl depletion did not induce significant changes in the locations or abundances of the {gamma}-subunit COOH terminus and CHIF in different kidney zones. We conclude that the unchanged levels of these two FXYD proteins suggest that they are not primary determinants for urine electrolyte composition during NaCl depletion.

Na-K-ATPase {gamma}-subunit; CHIF; kidney medulla



Address for reprint requests and other correspondence: A. B. Maunsbach, The Water and Salt Research Ctr, Institute of Anatomy Univ. of Aarhus, DK-8000 Aarhus, Denmark (e-mail: Maunsbach{at}ana.au.dk)




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