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Immunolocalization of Na/SO4-cotransport (NaSi-1) in rat kidney.


Lötscher, M; Custer, M; Quabius, E S; Kaissling, B; Murer, H; Biber, J (1996). Immunolocalization of Na/SO4-cotransport (NaSi-1) in rat kidney. Pflügers Archiv: European Journal of Physiology (Pflugers Archiv), 432(3):373-378.

Abstract

The proximal tubule is the major site for renal reabsorption of sulphate. A sodium-dependent transport system for sulphate (NaSi-1) has recently been identified from a rat kidney cortex cDNA library. Recent work demonstrated that NaSi-1 mRNA is expressed predominantly in proximal tubules. In the present work expression along the nephron of the Na/SO4-cotransporter NaSi-1 was studied by immunofluorescence. A polyclonal antibody was raised in rabbits against a fusion protein containing a 53-amino-acid polypeptide specific for the NaSi-1 sequence. The anti-NaSi-1 polyclonal antibody specifically detected a 68-kDa protein on Western blots and, by immunofluorescence specific staining, was observed in MDCK cells transfected with the NaSi-1 cotransporter. Using rat kidney cortex slices specific NaSi-1-related immunoreactivity was detected in proximal tubules and was restricted to the apical membrane. No immunoreactivity was observed in the other nephron segments. This was confirmed by Western blot analysis using proximal tubular apical and basolateral membranes isolated by free-flow electrophoresis. The results indicate that the Na/SO4-cotransporter NaSi-1 is expressed in the apical membrane of proximal tubular cells and is therefore likely to be involved in proximal reabsorption of sulphate.

The proximal tubule is the major site for renal reabsorption of sulphate. A sodium-dependent transport system for sulphate (NaSi-1) has recently been identified from a rat kidney cortex cDNA library. Recent work demonstrated that NaSi-1 mRNA is expressed predominantly in proximal tubules. In the present work expression along the nephron of the Na/SO4-cotransporter NaSi-1 was studied by immunofluorescence. A polyclonal antibody was raised in rabbits against a fusion protein containing a 53-amino-acid polypeptide specific for the NaSi-1 sequence. The anti-NaSi-1 polyclonal antibody specifically detected a 68-kDa protein on Western blots and, by immunofluorescence specific staining, was observed in MDCK cells transfected with the NaSi-1 cotransporter. Using rat kidney cortex slices specific NaSi-1-related immunoreactivity was detected in proximal tubules and was restricted to the apical membrane. No immunoreactivity was observed in the other nephron segments. This was confirmed by Western blot analysis using proximal tubular apical and basolateral membranes isolated by free-flow electrophoresis. The results indicate that the Na/SO4-cotransporter NaSi-1 is expressed in the apical membrane of proximal tubular cells and is therefore likely to be involved in proximal reabsorption of sulphate.

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Additional indexing

Item Type:Journal Article, refereed
Communities & Collections:04 Faculty of Medicine > Institute of Physiology
07 Faculty of Science > Institute of Physiology
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:1 July 1996
Deposited On:11 Feb 2008 12:22
Last Modified:05 Apr 2016 12:18
Publisher:Springer
ISSN:0031-6768
Publisher DOI:10.1007/s004240050147
Related URLs:http://www.springerlink.com/content/uw6dmq0hybq5nrbm/
PubMed ID:8765995

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