Skip to main content
Log in

Pivotal Role of Reduced Glutathione in Oxygen-induced Regulation of the Na+/K+ Pump in Mouse Erythrocyte Membranes

  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Abstract

This study addresses the mechanisms of oxygen-induced regulation of ion transport pathways in mouse erythrocyte, specifically focusing on the role of cellular redox state and ATP levels. Mouse erythrocytes possess Na+/K+ pump, K+-Cl and Na+-K+-2Cl cotransporters that have been shown to be potential targets of oxygen. The activity of neither cotransporter changed in response to hypoxia-reoxygenation. In contrast, the Na+/K+ pump responded to hypoxic treatment with reversible inhibition. Hypoxia-induced inhibition was abolished in Na+-loaded cells, revealing no effect of O2 on the maximal operation rate of the pump. Notably, the inhibitory effect of hypoxia was not followed by changes in cellular ATP levels. Hypoxic exposure did, however, lead to a rapid increase in cellular glutathione (GSH) levels. Decreasing GSH to normoxic levels under hypoxic conditions abolished hypoxia-induced inhibition of the pump. Furthermore, GSH added to the incubation medium was able to mimic hypoxia-induced inhibition. Taken together these data suggest a pivotal role of intracellular GSH in oxygen-induced modulation of the Na+/K+ pump activity.

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.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. N.C. Adragna P.K. Lauf (1997) ArticleTitleOxidative activation of K-Cl cotransport by diamide in erythrocytes from humans with red cell disorders, and from several other mammalian species. J. Membrane Biol. 155 207–217 Occurrence Handle1:CAS:528:DyaK2sXhsFGmtb4%3D

    CAS  Google Scholar 

  2. N.C. Adragna P.K. Lauf (1998) ArticleTitleRole of nitrite, a nitric oxide derivative, in K-Cl cotransport activation of low-potassium sheep red blood cells. J. Membrane Biol. 166 157–167 Occurrence Handle1:CAS:528:DyaK1MXhsFei

    CAS  Google Scholar 

  3. N. Akaike Y.H. Jin S. Koyama (1997) ArticleTitleFunctional interaction between Na+-K+ pump and KATP channels in the CNS neurons under experimental brain ischemia. Jpn. J. Physiol. 47 S50–S51 Occurrence Handle9266329

    PubMed  Google Scholar 

  4. D.J. Benos (1980) ArticleTitleIntracellular analysis of sodium, potassium, and chloride in mouse erythrocytes. J. Cell Physiol. 105 185–187 Occurrence Handle1:CAS:528:DyaL3MXisFai Occurrence Handle7430265

    CAS  PubMed  Google Scholar 

  5. D.J. Benos D.C. Tosteson (1980) ArticleTitleCation movements across mouse red blood cells. Biochim. Biophys. Acta 601 167–179 Occurrence Handle1:CAS:528:DyaL3cXls1Ghsbs%3D Occurrence Handle7407161

    CAS  PubMed  Google Scholar 

  6. M. Berenbrink S. Volkel N. Heisler M. Nikinmaa (2000) ArticleTitleO2-dependent K+ fluxes in trout red blood cells: the nature of O2 sensing revealed by the O2 affinity, cooperativity and pH dependence of transport. J. Physiol. 526 69–80 Occurrence Handle1:CAS:528:DC%2BD3cXlsFalsb0%3D Occurrence Handle10878100

    CAS  PubMed  Google Scholar 

  7. A.Y. Bogdanova M. Nikinmaa (2001) ArticleTitleReactive oxygen species regulate oxygen-sensitive potassium flux in rainbow trout erythrocytes. J. Gen. Physiol. 117 181–190 Occurrence Handle1:CAS:528:DC%2BD3MXht1Wjurw%3D Occurrence Handle11158169

    CAS  PubMed  Google Scholar 

  8. A.A. Boldyrev E.R. Bulygina (1997) ArticleTitleNa/K-ATPase and oxidative stress. Ann. N. Y. Acad. Sci. 834 666–668 Occurrence Handle1:CAS:528:DyaK1cXitl2jtA%3D%3D Occurrence Handle9405887

    CAS  PubMed  Google Scholar 

  9. U. Bruer M.K. Weih N.K. Isaev A. Meisel K. Ruscher A. Bergk G. Trendelenburg F. Wiegand I.V. Victorov U. Dirnagl (1997) ArticleTitleInduction of tolerance in rat cortical neurons: hypoxic preconditioning. FEBS Lett. 414 117–121 Occurrence Handle1:CAS:528:DyaK2sXlslCitL4%3D Occurrence Handle9305743

    CAS  PubMed  Google Scholar 

  10. L.T. Buck P.W. Hochachka (1993) ArticleTitleAnoxic suppression of Na+-K+-ATPase and constant membrane potential in hepatocytes: support for channel arrest. Am. J. Physiol. 265 R1020–R1025 Occurrence Handle1:CAS:528:DyaK2cXhtValt7Y%3D Occurrence Handle8238602

    CAS  PubMed  Google Scholar 

  11. E.H. Campbell J.S. Gibson (1998) ArticleTitleOxygen-dependent K+-fluxes in sheep red cells. J. Physiol. 506 679–688 Occurrence Handle1:CAS:528:DyaK1cXhtFSksbw%3D Occurrence Handle9503330

    CAS  PubMed  Google Scholar 

  12. R. Carini M.G. De Cesaris R. Splendore M. Bagnati E. Albano (2000) ArticleTitleIschemic preconditioning reduces Na+ accumulation and cell killing in isolated rat hepatocytes exposed to hypoxia. Hepatology 31 166–172 Occurrence Handle1:CAS:528:DC%2BD3cXks1OhsA%3D%3D Occurrence Handle10613742

    CAS  PubMed  Google Scholar 

  13. N.S. Chandel P.T. Schumacker (2000) ArticleTitleCellular oxygen sensing by mitochondria: old questions, new insight. J. Appl. Physiol. 88 1880–1889 Occurrence Handle1:CAS:528:DC%2BD3cXjvFSks70%3D Occurrence Handle10797153

    CAS  PubMed  Google Scholar 

  14. R.L. Colebrooke I.F. Smith P.J. Kemp C. Peers (2002) ArticleTitleChronic hypoxia remodels voltage-gated Ca2+ entry in a human airway chemoreceptor cell line. Neurosci. Lett. 318 69–72 Occurrence Handle1:CAS:528:DC%2BD38XktFaqtA%3D%3D Occurrence Handle11796188

    CAS  PubMed  Google Scholar 

  15. M. Comporti (2002) ArticleTitleIntroduction-serial review: iron and cellular redox status. Free Radic. Biol. Med. 32 565–567 Occurrence Handle1:CAS:528:DC%2BD38XitFWlsb8%3D Occurrence Handle11909690

    CAS  PubMed  Google Scholar 

  16. M. Cutaia N. Parks (1994) ArticleTitleOxidant stress decreases Na+/H+ antiport activity in bovine pulmonary artery endothelial cells. Am. J. Physiol. 261 L649–L659

    Google Scholar 

  17. D. Dobrota M. Matejovicova E.G. Kurella A.A. Boldyrev (1999) ArticleTitleNa/K-ATPase under oxidative stress: molecular mechanisms of injury. Cell. Mol. Neurobiol. 19 141–149 Occurrence Handle1:CAS:528:DyaK1MXhsFSitLg%3D Occurrence Handle10079973

    CAS  PubMed  Google Scholar 

  18. C. Domenicotti D. Paola A. Vitali M. Nitti C. d'Abramo D. Cottalasso G. Maloberti F. Biasi G. Poli E. Chiarpotto U.M. Marinari M.A. Pronzato (2000) ArticleTitleGlutathione depletion induces apoptosis of rat hepatocytes through activation of protein kinase C novel isoforms and dependent increase in AP-1 nuclear binding. Free Radic. Biol. Med. 29 1280–1290 Occurrence Handle1:CAS:528:DC%2BD3cXoslGnsrY%3D Occurrence Handle11118818

    CAS  PubMed  Google Scholar 

  19. E. Feraille M.L. Carranza B. Buffin-Meyer M. Rousselot A. Doucet H. Favre (1995) ArticleTitleProtein kinase C-dependent stimulation of Na+-K+-ATP epsilon in rat proximal convoluted tubules. Am. J. Physiol. 268 C1277–C1283 Occurrence Handle1:CAS:528:DyaK2MXlvVajtb0%3D Occurrence Handle7762622

    CAS  PubMed  Google Scholar 

  20. R. Ferrari C. Ceconi S. Curello A. Cargnoni O. Alfieri A. Pardini P. Marzollo O. Visioli (1991) ArticleTitleOxygen free radicals and myocardial damage: protective role of thiol-containing agents. Am. J. Med. 91 95S–105S Occurrence Handle1:CAS:528:DyaK38XltVKgsQ%3D%3D

    CAS  Google Scholar 

  21. H. Fujise K. Higa T. Kanemaru M. Fukuda N.C. Adragna P.K. Lauf (2001) ArticleTitleGSH depletion, K-Cl cotransport, and regulatory volume decrease in high-K/high-GSH dog red blood cells. Am. J. Physiol. 281 C2003–C2009 Occurrence Handle1:CAS:528:DC%2BD3MXptlKmsLs%3D

    CAS  Google Scholar 

  22. J.S. Gibson A.R. Cossins J.C. Ellory (2000) ArticleTitleOxygen-sensitive membrane transporters in vertebrate red cells. J. Exp. Biol. 203 1395–1407 Occurrence Handle1:CAS:528:DC%2BD3cXktF2gurk%3D Occurrence Handle10751155

    CAS  PubMed  Google Scholar 

  23. O.W. Griffith A. Meister (1985) ArticleTitleOrigin and turnover of mitochondrial glutathione. Proc. Natl. Acad. Sci. USA 82 4668–4672 Occurrence Handle1:CAS:528:DyaL2MXltVamsb8%3D Occurrence Handle3860816

    CAS  PubMed  Google Scholar 

  24. P.S. Haddock B. Woodward D.J. Hearse (1995) ArticleTitleCardiac Na+/K+-ATPase activity and its relation to myocardial glutathione status: studies in the rat. J. Mol. Cell Cardiol. 27 1185–1194 Occurrence Handle1:CAS:528:DyaK2MXlvFSitr8%3D Occurrence Handle7473776

    CAS  PubMed  Google Scholar 

  25. P.W. Hochachka P.L. Lutz (2001) ArticleTitleMechanism, origin, and evolution of anoxia tolerance in animals. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 130 435–459 Occurrence Handle1:STN:280:DC%2BD3MnktVGqtg%3D%3D Occurrence Handle11691622

    CAS  PubMed  Google Scholar 

  26. M. Inoue N. Fujishiro I. Imanaga (1999) ArticleTitleNa+ pump inhibition and non-selective cation channel activation by cyanide and anoxia in guinea-pig chromaffin cells. J. Physiol. 519 385–396 Occurrence Handle1:CAS:528:DyaK1MXmtlaqsr0%3D Occurrence Handle10457057

    CAS  PubMed  Google Scholar 

  27. M. Inoue N. Fujishiro I. Imanaga (2000) ArticleTitleRetardation of cation channel deactivation by mitochondrial dysfunction in adrenal medullary cells. Am. J. Physiol. 278 C26–C32 Occurrence Handle1:CAS:528:DC%2BD3cXht1Gjur0%3D

    CAS  Google Scholar 

  28. D. Kaji (1986) ArticleTitleVolume-sensitive K transport in human erythrocytes. J. Gen. Physiol. 88 719–738 Occurrence Handle1:CAS:528:DyaL2sXmsVyqug%3D%3D Occurrence Handle3794638

    CAS  PubMed  Google Scholar 

  29. E. Kurella M. Kukley O. Tyulina D. Dobrota M. Matejovicova V. Mezesova A. Boldyrev (1997) ArticleTitleKinetic parameters of Na/K-ATPase modified by free radicals in vitro and in vivo. Ann. N. Y. Acad. Sci. 834 661–665 Occurrence Handle1:CAS:528:DyaK1cXitl2jtw%3D%3D Occurrence Handle9405886

    CAS  PubMed  Google Scholar 

  30. P.K. Lauf N.C. Adragna N.S. Agar (1995) ArticleTitleGlutathione removal reveals kinases as common targets for K-Cl cotransport stimulation in sheep erythrocytes. Am. J. Physiol. 269 C234–C241 Occurrence Handle1:CAS:528:DyaK2MXnt1Gitb0%3D Occurrence Handle7631750

    CAS  PubMed  Google Scholar 

  31. J. Lopez-Barneo R. Pardal P. Ortega-Saenz (2001) ArticleTitleCellular mechanism of oxygen sensing. Annu. Rev. Physiol. 63 259–287 Occurrence Handle1:CAS:528:DC%2BD3MXjtFKmtLs%3D Occurrence Handle11181957

    CAS  PubMed  Google Scholar 

  32. H. Mairbaurl K. Mayer K.J. Kim Z. Borok P. Bartsch E.D. Crandall (2002) ArticleTitleHypoxia decreases active Na transport across primary rat alveolar epithelial cell monolayers. Am. J. Physiol. 282 L659–L665 Occurrence Handle1:CAS:528:DC%2BD38XivVeltrc%3D

    CAS  Google Scholar 

  33. H. Mairbaurl R. Wodopia S. Eckes S. Schulz P. Bartsch (1997) ArticleTitleImpairment of cation transport in A549 cells and rat alveolar epithelial cells by hypoxia. Am. J. Physiol. 273 L797–L806 Occurrence Handle1:CAS:528:DyaK2sXnt1WitLo%3D Occurrence Handle9357855

    CAS  PubMed  Google Scholar 

  34. R.M. Miller H. Sies E.M. Park J.A. Thomas (1990) ArticleTitlePhosphorylase and creatine kinase modification by thiol-disulfide exchange and by xanthine oxidase-initiated S-thiolation. Arch. Biochem. Biophys. 276 355–363 Occurrence Handle1:CAS:528:DyaK3cXhtlGjt7c%3D Occurrence Handle2106288

    CAS  PubMed  Google Scholar 

  35. M. Nikinmaa (2002) ArticleTitleOxygen-dependent cellular functions—why fishes and their aquatic environment are a prime choice of study. Comp. Biochem. Physiol. A Mol. Integr. Physiol. 133 1 Occurrence Handle12160868

    PubMed  Google Scholar 

  36. M. Nikinmaa A.Y. Bogdanova T. Lecklin (2003) ArticleTitleOxygen dependency of the adrenergic Na/H exchange in rainbow trout erythrocytes is diminished by a hydroxyl radical scavenger. Acta. Physiol. Scand. 178 1–6

    Google Scholar 

  37. H.L. Reeve E. Michelakis D.P. Nelson E.K. Weir S.L. Archer (2001) ArticleTitleAlterations in a redox oxygen sensing mechanism in chronic hypoxia. J. Appl. Physiol. 90 2249–2256 Occurrence Handle1:CAS:528:DC%2BD3MXksFansL8%3D Occurrence Handle11356790

    CAS  PubMed  Google Scholar 

  38. H. Sies A.L. Dafre Y. Ji T.P. Akerboom (1998) ArticleTitleProtein S-thiolation and redox regulation of membrane-bound glutathione transferase. Chem. Biol. Interact. 111-112 177–185 Occurrence Handle1:CAS:528:DyaK1cXitlSksbw%3D Occurrence Handle9679553

    CAS  PubMed  Google Scholar 

  39. I. Silver M. Erecinska (1998) ArticleTitleOxygen and ion concentrations in normoxic and hypoxic brain cells. Adv. Exp. Med. Biol. 454 7–16 Occurrence Handle1:CAS:528:DC%2BD3cXitVOns74%3D Occurrence Handle9889871

    CAS  PubMed  Google Scholar 

  40. B.Z. Simkhovich S. Abdishoo C. Poizat S.L. Hale L.H. Kedes R.A. Kloner (2002) ArticleTitleGene activity changes in ischemically preconditioned rabbit heart gene: discovery array study. Heart Dis. 4 63–69 Occurrence Handle1:CAS:528:DC%2BD38Xjs1GqsLY%3D Occurrence Handle11975836

    CAS  PubMed  Google Scholar 

  41. B.F. Sukhomlinov G.L. Antoniak B.O. Ikpu (1990) ArticleTitle[The effect of hypoxic hypoxia on the activity of glycolysis enzymes in rat erythrocytes] Ukr. Biokhim. Zh. 62 90–93 Occurrence Handle1:CAS:528:DyaK3cXlt12ru78%3D

    CAS  Google Scholar 

  42. K.J. Sweadner (1989) ArticleTitleIsozymes of the Na+/K+-ATPase. Biochim. Biophys. Acta 988 185–220 Occurrence Handle2541792

    PubMed  Google Scholar 

  43. E. Tanaka S. Yamamoto Y. Kudo S. Mihara H. Higashi (1997) ArticleTitleMechanisms underlying the rapid depolarization produced by deprivation of oxygen and glucose in rat hippocampal CA1 neurons in vitro. J. Neurophysiol. 78 891–902 Occurrence Handle9307122

    PubMed  Google Scholar 

  44. F. Tietze (1969) ArticleTitleEnzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal. Biochem. 27 502–522 Occurrence Handle1:CAS:528:DyaF1MXktVSlt7k%3D Occurrence Handle4388022

    CAS  PubMed  Google Scholar 

  45. A. Ziegelhoffer K. Kjeldsen H. Bundgaard A. Breier N. Vrbjar A. Dzurba (2000) ArticleTitleNa,K-ATPase in the myocardium: molecular principles, functional and clinical aspects. Gen. Physiol. Biophys. 19 9–47 Occurrence Handle1:CAS:528:DC%2BD3cXltFSiu7s%3D Occurrence Handle10930137

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by Swiss National Science Foundation (No 31-56743.99) for M.Gassmann, Olga Mayenfisch scholarship for A.Bogdanova and EMDO-scholarship for O.Ogunshola. We also thank Thomas Hofer for useful comments on the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to A. Y. Bogdanova.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bogdanova, A., Ogunshola, O., Bauer, C. et al. Pivotal Role of Reduced Glutathione in Oxygen-induced Regulation of the Na+/K+ Pump in Mouse Erythrocyte Membranes . J. Membrane Biol. 195, 33–42 (2003). https://doi.org/10.1007/s00232-003-2042-8

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00232-003-2042-8

Keywords

Navigation