Skip to main content
SpringerLink
Log in

The use of ionophores of rapid loading of human red cells with radioactive cations for cation-pump studies

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

Summary

Techniques are described for the rapid loading of intact human red cells with radioactive isotopes of alkali cations or Ca2+ by using ionophorous compounds (nigericin, gramicidin D and A 23187). Loading was rapid and efficient if the membrane potential of the cells was rendered more negative inside. After cation loading the ionophores could be bound to albumin and removed by repeated washings. The ATP and 2,3-DPG contents of the cells were practically unaltered by this treatment. Passive membrane permeability to Na+ and Ca2+ returned to normal. Loaded erythrocytes pumped out Na+ in a ouabain-sensitive and Ca2+ in a lanthanum-sensitive way. Ca2+-loaded red cells were microspherocytes and exhibited a rapid K+-efflux. Parallel with the extrusion of Ca2+ cells regained their biconcave shape and normal passive permeability to K+.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bodemann, H., Passow, H. 1972. Factors controlling the resealing of the membrane of human erythrocyte ghosts after hypotonic hemolysis.J. Membrane Biol. 8:1

    Google Scholar 

  • Cass, A., Dalmark, M. 1973. Equilibrium dialysis of ions in nystatin-treated red cells.Nature, New Biol. 244:47

    Google Scholar 

  • Dunn, M.J. 1974. Red blood cell calcium and magnesium: Effects upon sodium and potassium transport and cellular morphology.Biochim. Biophys. Acta 352:97

    PubMed  Google Scholar 

  • Ekman, A., Manninen, V., Salminen, S. 1969. Ion movements in red cells treated with propranolol.Acta Physiol. scand. 75:333

    PubMed  Google Scholar 

  • Gárdos, G. 1972. Ion transport across the erythrocyte membrane.Haematologia 6:237

    PubMed  Google Scholar 

  • Gárdos, G., Sarkadi, B., Szász, I. 1975. Effect of the Ca-ionophore A 23187 on the K-transport of human red cells.Abstr. Vth Int. Biophys. Congr. p. 100

  • Gárdos, G., Szász, I., Árky, I. 1966. Structure and function of erythrocytes. I. Relation between the energy metabolism and the maintenance of biconcave shape of human erythrocytes.Acta Biochim. Biophys. Acad. Sci. Hung. 1:253

    Google Scholar 

  • Garrahan, P.J., Glynn, I.M. 1967. The behaviour of the sodium pump in red cells in the absence of external potassium.J. Physiol. 192:159

    PubMed  Google Scholar 

  • Garrahan, P.J., Rega, A.F. 1967. Cation loading of red blood cells.J. Physiol. 193:459

    PubMed  Google Scholar 

  • Glynn, I.M. 1956. Sodium and potassium movements in human red cells.J. Physiol. 134:278

    PubMed  Google Scholar 

  • Glynn, I.M., Warner, A.E. 1972. Nature of the calcium dependent potassium leak induced by (+)-propranolol, and its possible relevance to the drug's antiarrhythmic effect.Brit. J. Pharmacol. 44:271

    Google Scholar 

  • Gunn, R.B., Tosteson, D.C. 1971. The effect of 2,4,6-trinitro-m-cresol on cation and anion transport in sheep red blood cells.J. Gen. Physiol. 57:593

    PubMed  Google Scholar 

  • Harris, E.J., Prankerd, T.A.J. 1953. The rate of sodium extrusion from human erythrocytes.J. Physiol. 121:470

    PubMed  Google Scholar 

  • Harrison, D.G., Long, C. 1968. The calcium content of human erythrocytes.J. Physiol. 199:367

    PubMed  Google Scholar 

  • Lee, K.S., Shin, B.Ch. 1969. Studies on the active transport of calcium in human red cells.J. Gen. Physiol. 54:713

    PubMed  Google Scholar 

  • Lew, V.L. 1974. On the mechanism of the Ca-induced increase in K permeability observed in human red cell membranes.In: Comparative Biochemistry and Physiology of Transport. L. Bolis, K. Bloch, S.E. Luria, and F. Lynen, editors. p. 310. North-Holland Publishing Co., Amsterdam-London

    Google Scholar 

  • Lichtman, M.A., Weed, R.J. 1973. Divalent cation content of normal and ATP-depleted erythrocytes and erythrocyte membranes.In: Red Cell Shape. M. Bessis, R.I. Weed, and P.F. Leblond, editors. p. 79. Springer-Verlag, New York

    Google Scholar 

  • Lubowitz, H., Whittam, R. 1969. Ion movements in human red cells independent of the sodium pump.J. Physiol. 202:111

    PubMed  Google Scholar 

  • Olson, E.J., Cazort, R.J. 1969. Active calcium and strontium transport in human erythrocyte ghosts.J. Gen. Physiol. 53:311

    PubMed  Google Scholar 

  • Quist, E.E., Roufogalis, B.D. 1975. Determination of the stoichiometry of the calcium pump in human erythrocytes using lanthanum as a selective inhibitor.FEBS Lett. 50:135

    PubMed  Google Scholar 

  • Reed, P.W. 1973. Calcium-dependent potassium efflux from rat erythrocytes incubated with antibiotic A 23187.Fed. Proc. 32:635 (Abstr.)

    Google Scholar 

  • Reed, P.W., Lardy, H.A. 1972. A23187: A divalent cation ionophore.J. Biol. Chem. 247:6970

    PubMed  Google Scholar 

  • Schatzmann, H.J. 1966. ATP-dependent Ca++ extrusion from human red cells.Experientia 22:364

    PubMed  Google Scholar 

  • Schatzmann, H.J. 1973. Dependence on calcium concentration and stoichiometry of the calcium pump in human red cells.J. Physiol. 235:551

    PubMed  Google Scholar 

  • Schatzmann, H.J., Tschabold, M. 1971. The lanthanides Ho3+ and Pr3+ as inhibitors of calcium transport in human red cells.Experientia 27:59

    PubMed  Google Scholar 

  • Schatzmann, H.J., Vincenzi, F.F. 1969. Calcium movements across the membrane of human red cells.J. Physiol. 201:369

    PubMed  Google Scholar 

  • Schwoch, G., Passow, H. 1973. Preparation and properties of human erythrocyte ghosts.Mol. Cell. Biochem. 2:197

    PubMed  Google Scholar 

  • Szász, I., Gárdos, G., Árky, I. 1971. Study of ATP-consuming functions of erythrocytes during blood preservation.Bibl. Haematol. (Basel) 38:196

    Google Scholar 

  • Weiss, G.B. 1974. Cellular pharmacology of lanthanum.Annu. Rev. Pharmacol. 14:343

    Google Scholar 

  • White, J.G. 1974. Effects of an ionophore, A23187, on the surface morphology of normal erythrocytes.Am. J. Pathol. 77:507

    PubMed  Google Scholar 

  • Whittam, R., Ager, M.E. 1965. The connexion between active cation transport and metabolism in erythrocytes.Biochem. J. 97:214

    Google Scholar 

  • Wolf, H.U. 1972. Studies on a Ca2+-dependent ATPase of human erythrocyte membranes. Effects of Ca2+ and H+.Biochim. Biophys. Acta 266:361

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell Metabolism, National Institute of Haematology and Blood Transfusion, Budapest, Hungary

    B. Sarkadi, Ilma Szász & G. Gárdos

Authors
  1. B. Sarkadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ilma Szász
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Gárdos
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sarkadi, B., Szász, I. & Gárdos, G. The use of ionophores of rapid loading of human red cells with radioactive cations for cation-pump studies. J. Membrain Biol. 26, 357–370 (1976). https://doi.org/10.1007/BF01868883

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01868883

Keywords

Search

Navigation