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

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 NaSi1-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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References

1. Biber J, Stieger B, Haase W, Murer H (1981) A high yield preparation of rat kidney brush border membranes. Different behaviour of lysosomal markers. Biochim Biophys Acta 647:169–176
   Article PubMed CAS Google Scholar
2. Busch AE, Waldegger S, Herzer T, Biber J, Markovich D, Murer H, Lang F (1994) Electrogenic cotransport of Na and sulfate in Xenopus laevis oocytes expressing the cloned Na/SO4 transport protein NaSi-1. J Biol Chem 269:12407–12409
   PubMed CAS Google Scholar
3. Custer M, Lötscher M, Biber J, Murer H, Kaissling B (1994) Expression of Na/Pi-cotransport (NaPi-2) in rat kidney: localization by RT-PCR and immunohistochemistry. Am J Physiol 266:F676-F774
   Google Scholar
4. Custer M, Murer H, Biber J (1994) Nephron localization of Na/SO4-cotransport related mRNA and protein. Pflügers Arch 429:165–168
   Article PubMed CAS Google Scholar
5. Dawson TP, Gandhi R, Le Hir M, Kaissling B (1989) Ecto-5′nucleotidase: localization by light microscopic histochemistry and immunohistochemistry methods in the rat kidney. J Histochem Cytochem 37:39–47
   PubMed CAS Google Scholar
6. Heidrich HG, Kinne R, Kinne-Saffran E, Hanig K (1972) The polarity of the proximal tubule cell in rat kidney. J Cell Biol 54:232–245
   Article PubMed CAS Google Scholar
7. Hierholzer K, Cade R, Gurd R, Kessler R, Pitts R (1960) Stop flow analysis of renal reabsorption and excretion of sulfate in the dog. Am J Physiol 198:833–837
   PubMed CAS Google Scholar
8. Hirt RPO, Poulain-Godefroy O, Billote J, Kraehenbuhl J-P, Fasel N (1992) Highly inducible synthesis of heterologous proteins in epithelial cells carrying glucocorticoid responsive vector. Gene 111:199–206
   Article PubMed CAS Google Scholar
9. Markovich D, Forgo J, Stange G, Biber J, Murer H (1993) Expression cloning of rat Na/SO4 cotransport. Proc Natl Acad Sci USA 90:8073–8077
   Article PubMed CAS Google Scholar
10. Murer H, Manganel M, Roch-Ramel F (1992) Tubular transport of monocarboxylates, Krebs-cycle intermediates and inorganic sulphate. In: Windhager E (ed) Handbook of physiology, vol 8. Oxford University Press, New York, pp 165–218
    Google Scholar
11. Pajor AM (1995) Expression cloning of a renal sodium/dicarboxylate cotransporter. J Biol Chem 270:5779–5785
    PubMed CAS Google Scholar
12. Quabius ES, Murer H, Biber J (1996) Expression of proximal tubular Na-Pi and Na-SO4 cotransporters in MDCK and LLC-PK1 cells by transfection. Am J Physiol 39: F220-F228
    Google Scholar
13. Ullrich KJ, Murer H (1982) Sulphate and phosphate transport in renal proximal tubule. Philos Trans R Soc Lond Biol 299:549–558
    Article PubMed CAS Google Scholar

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Author notes

1. J. Biber
   Present address: Institute of Physiology, University Zürich-Irchel, Winterthurerstrasse 190, CH-8057, Zürich, Switzerland

Authors and Affiliations

1. Institute of Physiology, University Zürich-Irchel, Winterthurerstrasse 190, CH-8057, Zürich, Switzerland
   M. Custer, E. S. Quabius & H. Murer
2. Institute of Anatomy, University of Zürich, Zürich, Switzerland
   M. Lötscher & B. Kaissling

Authors

1. J. Biber
2. M. Custer
3. E. S. Quabius
4. H. Murer
5. M. Lötscher
6. B. Kaissling

Additional information

M. Lötscher and M. Custer contributed equally to this work.

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Biber, J., Custer, M., Quabius, E.S. et al. Immunolocalization of Na/SO4-cotransport (NaSi-1) in rat kidney.Pflügers Arch — Eur J Physiol 432, 373–378 (1996). https://doi.org/10.1007/s004240050147

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• Received: 31 January 1996
• Accepted: 21 March 1996
• Issue date: July 1996
• DOI: https://doi.org/10.1007/s004240050147

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