Skip to main content

Advertisement

Log in

Expression of the human sodium/proton exchanger NHE-1 in Xenopus laevis oocytes enhances sodium/proton exchange activity and establishes sodium/lithium countertransport

  • Original Article
  • Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

We investigated whether the human sodium/proton (Na+/H+) exchanger isoform 1 (NHE-1) can mediate sodium/lithium (Na+/Li+) coutertransport. Using the Xenopus laevis oocyte expression system we determined amiloride-sensitive Li+ uptake, a measure of Na+/H+ exchange, in oocytes injected with water or NHE-1 cRNA. Amiloride-sensitive Li+ uptake was three-to tenfold enhanced over control in NHE-1 cRNA-injected cells and was selectively inhibited by 0.01 μM HOE 694 [i.e. (3-methylsulphonyl-4-piperidinobenzoyl) guanidine methanesulphonate]. The endogenously present Na+/H+ exchanger was insensitive to HOE 694. After acidification of oocytes from pH 7.7 to 6.8, amiloride-sensitive Li+ uptake was four-to tenfold higher in NHE-1 cRNA-injected cells than in controls. Li+ efflux from control oocytes was independent of extracellular Na+, indicating that these cells expressed no measurable Na+/Li+ countertransport activity. In NHE-1 cRNA-injected oocytes, Li+ efflux was distinctly enhanced by extracellular Na+ ions. This Na+-dependent Li+ efflux was inhibited by ethylisopropylamiloride, phloretin and by cytosolic acidification. The data show that expression of the NHE-1 in X. laevis oocytes induces the expression of Na+/Li+ countertransport. The data confirm that Na+/H+ exchange and Na+/Li+ countertransport are mediated by the same transport system.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Aronson PS (1985) Kinetic properties of the plasma membrane Na+/H+ exchanger. Annu Rev Physiol 47:545–560

    Google Scholar 

  2. Borgese F, Sardet C, Cappadoro M, Pouysségur J, Motais R (1992) Cloning and expression of a cAMP-activated Na+/H+ exchanger:evidence that the cytoplasmic domaine mediates hormonal regulation. Proc Natl Acad Sci USA 89:6765–6769

    Google Scholar 

  3. Burckhardt B-C, Frömter E (1992) Pathways of NH3/ +4 permeation across Xenopus laevis oocyte cell membrane. Pflügers Arch 420:83–86

    Google Scholar 

  4. Burckhardt B-C, Kroll B, Frömter E (1992) Proton transport mechanism in the cell membrane of Xenopus laevis oocytes. Pflügers Arch 420:78–82

    Google Scholar 

  5. Canessa M (1989) Kinetic properties of Na+/H+ exchange and Li+/Na+, Na+/Na+, and Na+/Li+ exchanges of human red cells. Methods Enzymol 173:176–191

    Google Scholar 

  6. Canessa M, Morgan K, Semplicini A (1988) Genetic differences in lithiumsodium exchange and regulation of the sodium-hydrogen exchanger in essential hypertension. J Cardiovasc Pharmacol 12 [Suppl 3]:S92-S98

    Google Scholar 

  7. Canessa M, Morgan K, Goldszer R, Moore TJ, Spalvins A (1991) Kinetic abnormalities of the red blood cell sodium-proton exchange in hypertensive patients. Hypertension 17:340–348

    Google Scholar 

  8. Cicirelli MF, Robinson KR, Smith LD (1983) Internal pH of Xenopus oocytes: a study on the mechanism and role of pH changes during meiotic maturation. Dev Biol 100:133–146

    Google Scholar 

  9. Corry DB, Tuck ML, Nicholas S, Weinman EJ (1993) Increased Na/H antiport activity and abundance in uremie red blood cells. Kidney Int 44:574–578

    Google Scholar 

  10. Counillon L, Scholz W, Lang HJ, Pouysségur J (1993) Pharmacological characterization of stably transfected Na+/H+ an-tiporter isoforms using amiloride analogs and a new inhibitor exhibiting anti-ischemic properties. Mol Pharmacol 44:1041–1045

    Google Scholar 

  11. Duhm J, Becker BF (1979) Studies on lithium transport across the red cell membrane. V. On the nature of the Na+-dependent Li+ countertransport system of mammalian erythrocytes. J Membr Biol 51:263–286

    Google Scholar 

  12. Escobales N, Canessa M (1986) Amiloride-sensitive Na+ transport in human red cells: evidence for a Na+/H+ exchange system. J Membr Biol 90:21–28

    Google Scholar 

  13. Escobales N, Figueroa J (1991) Na+/Na+ exchange and Na+/H+ antiport in rabbit erythrocytes: two distinct transport systems. J Membr Biol 120:41–49

    Google Scholar 

  14. Fliegel L, Fröhlich O (1993) The Na+/H+ exchanger: an update on structure, regulation and cardiac physiology. Biochem J 296:273–285

    Google Scholar 

  15. Franchi A, Perrucca-Lostanlen D, Pouysségur J (1986) Functional expression of a human Na+/H+ antiporter gene transfected into antiporter-deficient mouse L cells. Proc Natl Acad Sci USA 83:9388–9392

    Google Scholar 

  16. Goldin AL (1992) Maintenance of Xenopus laevis and oocyte injection. Methods Enzymol 207:266–279

    Google Scholar 

  17. Guizouarn H, Borgese F, Pellissier B, Garcia-Romeu F, Motais R (1993) Role of protein phophorylation and dephosphorylation in activation and desensitization of the cAMP-dependent Na+/H+ antiport. J Biol Chem 268:8632–8639

    Google Scholar 

  18. Huot SJ, Aronson PS (1991) Na+-H+ exchanger and its role in essential hypertension and diabetes mellitus. Diabetes Care 14:521–535

    Google Scholar 

  19. Kahn AM, Allen JC, Cragoe EJ Jr, Shelat H (1989) Sodium-lithium exchange and sodium-proton exchange are mediated by the same transport system in sarcolemmal vesicles from bovine superior mesenteric artery. Circ Res 65:818–828

    Google Scholar 

  20. Krolewski AS, Canessa M, Warram JH, Laffel LMB, Christlieb AR, Knowler WC, Rand LI (1988) Predisposition to hypertension and susceptibility to renal disease in insulin-dependent diabetes mellitus. N Engl J Med 318:140–145

    Google Scholar 

  21. Lee SC, Steinhardt RA (1981) pH changes associated with meiotic maturation in oocytes of Xenopus laevis. Dev Biol 85:358–369

    Google Scholar 

  22. Levine SA, Montrose MH, Tse CM, Donowitz M (1993) Kinetics and regulation of three cloned mammalian Na+/H+ exchangers stably expressed in a fibroblast cell line. J Biol Chem 268:25527–25535

    Google Scholar 

  23. Mangili R, Bending JJ, Scott G, Li LK, Gupta A, Viberti G (1988) Increased sodium-lithium countertransport activity in red cells of patients with insulindependent diabetes and nephropathy. N Engl J Med 318:145–150

    Google Scholar 

  24. Merritt JE, Armstrong JE, Benham CD, Hallam TJ, Jacob R, Jaxa-Chamiec A, Leigh BK, McCarthy SA, Moores KE, Rink TJ (1990) SK&F 96365, a novel inhibitor of receptor-mediated calcium entry. Biochem J 271:515–522

    Google Scholar 

  25. Morgan K, Canessa M (1990) Interactions of external and internal H+ and Na+ with Na+/Na+ and Na+/H+ exchange of rabbit red cells:evidence for a common pathway. J Membr Biol 118:193–214

    Google Scholar 

  26. Orlowski J (1993) Heterologous expression and functional properties of amiloride high affinity (NHE-1) and low affinity (NHE-3) isoforms of the rat Na/H exchanger. J Biol Chem 268:16369–16377

    Google Scholar 

  27. Pouysségur J, Sardet C, Franchi A, L'Allemain G, Paris S (1984) A specific mutation abolishing Na+/H+ antiport activity in hamster fibroblasts precludes growth at neutral and acidic pH. Proc Natl Acad Sci USA 81:4833–4837

    Google Scholar 

  28. Rosskopf D, Düsing R, Siffert W (1993) Membrane sodium-proton exchange and primary hypertension. Hypertension 21:607–617

    Google Scholar 

  29. Rosskopf D, Frömter E, Siffert W (1993) Hypertensive sodium-proton exchanger phenotype persists in immortalized lymphoblasts from essential hypertensive patients — a cell culture model for human hypertension. J Clin Invest 92:2553–2559

    Google Scholar 

  30. Sardet C, Franchi A, Pouysségur J (1989) Molecular cloning, primary structure, and expression of the human growth factor-activalable Na+/H+ antiporter. Cel 156:271–280

    Google Scholar 

  31. Sasaki S, Ishibashi K, Nagai T, Marumo F (1992) Regulation mechanisms of intracellular pH of Xenopus laevis oocyte. Biochim Biophys Acta 1137:45–51

    Google Scholar 

  32. Towle DW, Baksinski A, Richard NE, Kordylewski M (1991) Characterization of an endogenous Na+/H+ antiporter in Xenopus laevis oocytes. J Exp Biol 159:359–369

    Google Scholar 

  33. Tse M, Levine S, Yun C, Brant S, Counillon LT, Pouyssegur J, Donowitz M (1993) Structure/function studies of the epithelial isoforms of the mammalian Na+/H+ exchanger gene family. J Membr Biol 135:93–108

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Busch, S., Burckhardt, B.C. & Siffert, W. Expression of the human sodium/proton exchanger NHE-1 in Xenopus laevis oocytes enhances sodium/proton exchange activity and establishes sodium/lithium countertransport. Pflügers Arch. 429, 859–869 (1995). https://doi.org/10.1007/BF00374811

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00374811

Key words

Navigation