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Am J Physiol Renal Physiol (June 6, 2006). doi:10.1152/ajprenal.00086.2006
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Submitted on March 14, 2006
Accepted on May 31, 2006

Locations, abundances and possible functions of FXYD ion transport regulators in rat renal medulla

Kaarina Pihakaski-Maunsbach1, Henrik Vorum2, Bent Honore2, Shigeki Tokonabe3, Jorgen Frokiaer4, Haim Garty5, Steven J.D. Karlish6, and Arvid B. Maunsbach7*

1 The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark
2 Institute of Medical Biochemistry, University of Aarhus, Aarhus, Denmark
3 The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark; Aarhus, United States
4 The Water and Salt Research Center, University of Aarhus, Denmark
5 Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel
6 Biological Chemistry, The Weizmann Institute of Science, Israel
7 The Water and Salt Research Center, Institute of Anatomy, University of Aarhus, Aarhus, Denmark; , Denmark

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

The {gamma} subunit of Na,K-ATPase (FXYD2) and CHIF (FXYD4) are considered pump regulators in kidney tubules. The aim of this study was to expand the information about their locations in 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 medulla of control and sodium depleted rats were analyzed by immunofluorescence and immunoelectron microscopy, and semiquantitative Western blotting. The results showed that antibodies against {gamma}C-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 {gamma}C-terminus, splice variant {gamma}a, and CHIF. Splice variant {gamma}b was abundant in inner stripe of outer medulla (ISOM) but absent in inner medulla (IM). Double labeling by high resolution immunoelectron microscopy showed close structural association between CHIF and 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 renal medulla and show a new {gamma} variant in inner medulla. Extensive sodium chloride depletion did not induce significant changes in the locations or abundances of {gamma}C-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 sodium chloride depletion.




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