Journal of Comparative Physiology B

, Volume 159, Issue 3, pp 339–347 | Cite as

Presence of a Na+/H+ exchanger in brush border membranes isolated from the kidney of the spiny dogfish,Squalus acanthias

  • C. Bevan
  • R. K. H. Kinne
  • R. E. Shetlar
  • E. Kinne-Saffran
Article

Summary

A membrane fraction, rich in brush border membranes, was prepared from renal proximal tubules of the spiny dogfish,Squalus acanthias, and the sodium-proton exchange mechanism in these membrane vesicles was investigated by both a rapid filtration technique and the fluorescence quenching of acridine organe.22Na+ uptake was stimulated by an outwardly directed H+ gradient, and was inhibited by amiloride at a single inhibitory site with an apparentK i of approximately 1.7×10−5M. In the presence of an H i + >H o + gradient, the\(K_{{\text{m}}_{{\text{Na}}} + } {\text{and}} V_{\max _{{\text{Na}}} + } \) of the Na+/H+ exchanger were 9.7±0.8 mM and 48.0±12.0 nmol·mg protein−1·min−1, respectively. The uptake of Na+ was electroneutral in the presence of a H+ gradient, indicating a stoichiometry of 1. In the fluorescence studies, quenching of acridine orange occurred in the presence of an outwardly directed Na+ gradient which was inhibited by amiloride. Thus, an electroneutral Na+/H+ exchanger with properties similar to those found in the mammalian kidney is also present in the spiny dogfish and may contribute to the urinary acidification of this marine animal.

Key words

Renal acidification Na+/H+ exchange Proximal tubule Amiloride Brush border membrane vesicles Squalus acanthias 

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References

  1. Aronson PS (1983) Mechanisms of active H+ secretion in the proximal tubule. Am J Physiol 245:F647-F659Google Scholar
  2. Aronson PS, Suhm MA, Nee J (1983) Interaction of external H+ with the Na+/H+ exchanger in renal microvillus membrane vesicles. J Biol Chem 258:6767–6771Google Scholar
  3. Booth AG, Kenny AJ (1974) A rapid method for the preparation of microvilli from rabbit kidney. Biochem J 142:575–581Google Scholar
  4. Borgers M, Thoné F (1975) The inhibition of alkaline phosphatase by L-p-bromotetramisole. Histochemistry 44:277–280Google Scholar
  5. Burnham C, Munzesheimer C, Rabon E, Sachs G (1982) Ion pathways in renal brush border membranes. Biochim Biophys Acta 685:260–272Google Scholar
  6. Cohn DE, Hruska KA, Klahr S, Hammerman MR (1982) Increased Na+−H+ exchange in brush border vesicles from dogs with renal failure. Am J Physiol 243:F293-F299Google Scholar
  7. Eveloff J, Kinne R, Kinter WB (1979) p-Aminohippuric acid transport into brush border vesicles isolated from flounder kidney. Am J Physiol 237:F291-F298Google Scholar
  8. Eveloff J, Field M, Kinne R, Murer R (1980) Sodium-cotransport systems in intestine and kidney of the winter flounder. J Comp Physiol 135:175–182Google Scholar
  9. Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorus. J Biol Chem 66:375–388Google Scholar
  10. Green DE, Ziegler DM (1963) Electron transport particles. In: Colowick SP, Kaplan NO (eds) Methods of enzymology, vol VI. Academic Press, New York, pp 416–424Google Scholar
  11. Greger R, Schlatter E, Wang F, Forrest JN, Jr (1984) Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). III. Effects of stimulation by cyclic AMP. Pflügers Arch 402:376–384Google Scholar
  12. Hentschel H (1987) Renal architecture of the dogfish (Scyliorhinus caniculus (Chondrichthyes, Elasmobranchii). Zoomorphology 107:115–125Google Scholar
  13. Hodler J, Heinemann HO, Fishman AP, Smith HW (1955) Urine pH and carbonic anhydrase activity in the marine dogfish. Am J Physiol 183:155–162Google Scholar
  14. Hoelzl Wallach DF, Kamat VB (1966) Preparation of plasma membrane fragments from mouse ascites tumor cells. In: Colowick SP, Kaplan NO (eds) Methods of enzymology, vol VIII. Academic Press, New York, pp 164–172Google Scholar
  15. Hopfer U, Nelson K, Perrotto J, Isselbacher KJ (1973) Glucose transport in isolated brush border membranes from rat small intestine. J Biol Chem 248:25–32Google Scholar
  16. Hugon J, Borgers M (1966) A direct lead method for the electron microscopic visualization of alkaline phosphatase activity. J Histochem Cytochem 14:429Google Scholar
  17. Kempton RT (1940) The site of acidification of urine within the renal tubule of the dogfish. Bull Mt Desert Isl Biol Lab, pp 34–36Google Scholar
  18. Kinne R (1985) Transport function of renal cell membranes: sodium cotransport systems. In: Kinne RKH (ed) Renal biochemistry. Cells, membranes, molecules. Elsevier, Amsterdam, pp 99–141Google Scholar
  19. Kinne R, Schmitz JE, Kinne-Saffran E (1971) The localization of the Na−K-ATPase in the cells of rat kidney cortex. A study on isolated plasma membranes. Pflügers Arch 329:191–206Google Scholar
  20. Kinsella JL, Aronson PS (1980) Properties of the Na+−H+ exchanger in renal microvillus membrane vesicles. Am J Physiol 238:F461-F469Google Scholar
  21. Kinsella JL, Aronson PS (1981) Amiloride inhibition of the Na+−H+ exchanger in renal microvillus membrane vesicles. Am J Physiol 241:F374-F379Google Scholar
  22. Kinsella JL, Aronson PS (1982) Determination of the coupling ratio for Na+−H+ exchange in renal microvillus vesicles. Biochim Biophys Acta 689:161–164Google Scholar
  23. Lansing AI, Belkhode ML, Lynch WE, Lieberman I (1967) Enzymes of plasma membranes of liver. J Biol Chem 242:1772–1775Google Scholar
  24. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  25. Maren TH (1988) Renal acidification in marine fish fifty years after Homer Smith. Proc Symp Renal, Fluid, Electrolyte Physiol, pp 28–35Google Scholar
  26. Murer H, Hopfer U, Kinne R (1976) Sodium/proton antiport in brush-border-membrane vesicles isolated from rat small intestine and kidney. Biochem J 154:597–604Google Scholar
  27. Nord EP, Hafezi A, Wright EM, Fine LG (1984) Mechanisms of Na+ uptake into renal brush border membrane vesicles. Am J Physiol 247:F548-F554Google Scholar
  28. Sabolić I, Burckhardt G (1983) Proton pathways in rat renal brush-border and basolateral membranes. Biochim Biophys Acta 734:210–220Google Scholar
  29. Seifter J, Kinsella JL, Aronson PS (1981) Mechanism of Cl transport inNecturus renal microvillus membrane vesicles. Kidney Int 19:257Google Scholar
  30. Smith WW (1939) The excretion of phosphate in the dogfish,Squalus acanthias. J Cell Comp Physiol 14:95–102Google Scholar
  31. Swenson ER, Maren TH (1986) Dissociation of CO2 hydration and renal acid secretion in the dogfish,Squalus acanthias. Am J Physiol 250:F288-F293Google Scholar
  32. Warnock DG, Reenstra WW, Yee VJ (1982) Na+/H+ antiporter of brush border vesicles: studies with acridine orange uptake. Am J Physiol 242:F733-F739Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • C. Bevan
    • 1
    • 2
  • R. K. H. Kinne
    • 1
    • 2
  • R. E. Shetlar
    • 1
    • 2
  • E. Kinne-Saffran
    • 1
    • 2
  1. 1.Max-Planck-Institut für SystemphysiologieDortmund 1Federal Republic of Germany
  2. 2.Mount Desert Island Biological LaboratorySalsbury CoveUSA

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