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. 1997 Jun 1;99(11):2727–2735. doi: 10.1172/JCI119462

Stochastic nature and red cell population distribution of the sickling-induced Ca2+ permeability.

V L Lew 1, O E Ortiz 1, R M Bookchin 1
PMCID: PMC508119  PMID: 9169503

Abstract

To explore basic properties of the sickling-induced cation permeability pathway, the Ca2+ component (Psickle-Ca) was studied in density-fractionated sickle cell anemia (SS) discocytes through its effects on the activity of the cells' Ca2+sensitive K+-channels (KCa). The instant state of KCa channel activation was monitored during continuous or cyclic deoxygenation of the cells using a novel thiocyanate-densecell formation method. Each deoxy pulse caused a reversible, sustained Psickle-Ca, which activated KCa channels in only 10-45% of cells at physiological [Ca2+]o ("activated cells"). After removal of cells activated by each previous deoxy pulse, subsequent pulses generated similar activated cell fractions, indicating a random determination rather than the response of a specific vulnerable subpopulation. The fraction of activated cells rose monotonically with [Ca2+]o along a curve reflecting the cells' distribution of Psickle-Ca, with values high enough in a small cell fraction to trigger near-maximal KCa channels. Consistent with the stochastic nature of Psickle-Ca, repeated deoxygenated-oxygenated pulsing led to progressive dense cell formation, whereas single long pulses caused one early density shift. Thus deoxygenation-induced Ca2+-permeabilization in SS cells is a probabilistic event with large cumulative dehydrating potential. The possible molecular nature of Psickle-Ca is discussed.

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Selected References

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  1. Allan D., Limbrick A. R., Thomas P., Westerman M. P. Release of spectrin-free spicules on reoxygenation of sickled erythrocytes. Nature. 1982 Feb 18;295(5850):612–613. doi: 10.1038/295612a0. [DOI] [PubMed] [Google Scholar]
  2. Bookchin R. M., Balazs T., Landau L. C. Determinants of red cell sickling. Effects of varying pH and of increasing intracellular hemoglobin concentration by osmotic shrinkage. J Lab Clin Med. 1976 Apr;87(4):597–616. [PubMed] [Google Scholar]
  3. Bookchin R. M., Ortiz O. E., Lew V. L. Evidence for a direct reticulocyte origin of dense red cells in sickle cell anemia. J Clin Invest. 1991 Jan;87(1):113–124. doi: 10.1172/JCI114959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bookchin R. M., Ortiz O. E., Somlyo A. V., Somlyo A. P., Sepulveda M. I., Hockaday A., Lew V. L. Calcium-accumulating inside-out vesicles in sickle cell anemia red cells. Trans Assoc Am Physicians. 1985;98:10–20. [PubMed] [Google Scholar]
  5. Cao Z., Ferrone F. A. A 50th order reaction predicted and observed for sickle hemoglobin nucleation. J Mol Biol. 1996 Feb 23;256(2):219–222. doi: 10.1006/jmbi.1996.0079. [DOI] [PubMed] [Google Scholar]
  6. Clark M. R., Guatelli J. C., Mohandas N., Shohet S. B. Influence of red cell water content on the morphology of sickling. Blood. 1980 May;55(5):823–830. [PubMed] [Google Scholar]
  7. Eaton J. W., Skelton T. D., Swofford H. S., Kolpin C. E., Jacob H. S. Elevated erythrocyte calcium in sickle cell disease. Nature. 1973 Nov 9;246(5428):105–106. doi: 10.1038/246105a0. [DOI] [PubMed] [Google Scholar]
  8. Etzion Z., Tiffert T., Bookchin R. M., Lew V. L. Effects of deoxygenation on active and passive Ca2+ transport and on the cytoplasmic Ca2+ levels of sickle cell anemia red cells. J Clin Invest. 1993 Nov;92(5):2489–2498. doi: 10.1172/JCI116857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ferreira H. G., Lew V. L. Use of ionophore A23187 to measure cytoplasmic Ca buffering and activation of the Ca pump by internal Ca. Nature. 1976 Jan 1;259(5538):47–49. doi: 10.1038/259047a0. [DOI] [PubMed] [Google Scholar]
  10. Fiévet B., Gabillat N., Borgese F., Motais R. Expression of band 3 anion exchanger induces chloride current and taurine transport: structure-function analysis. EMBO J. 1995 Nov 1;14(21):5158–5169. doi: 10.1002/j.1460-2075.1995.tb00200.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. García-Sancho J., Lew V. L. Detection and separation of human red cells with different calcium contents following uniform calcium permeabilization. J Physiol. 1988 Dec;407:505–522. doi: 10.1113/jphysiol.1988.sp017428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glynn I. M., Warner A. E. Nature of the calcium dependent potassium leak induced by (+)-propranolol, and its possible relevance to the drug's antiarrhythmic effect. Br J Pharmacol. 1972 Feb;44(2):271–278. doi: 10.1111/j.1476-5381.1972.tb07263.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Joiner C. H. Deoxygenation-induced cation fluxes in sickle cells: II. Inhibition by stilbene disulfonates. Blood. 1990 Jul 1;76(1):212–220. [PubMed] [Google Scholar]
  14. Lew V. L., Hockaday A., Sepulveda M. I., Somlyo A. P., Somlyo A. V., Ortiz O. E., Bookchin R. M. Compartmentalization of sickle-cell calcium in endocytic inside-out vesicles. Nature. 1985 Jun 13;315(6020):586–589. doi: 10.1038/315586a0. [DOI] [PubMed] [Google Scholar]
  15. Lew V. L., Tsien R. Y., Miner C., Bookchin R. M. Physiological [Ca2+]i level and pump-leak turnover in intact red cells measured using an incorporated Ca chelator. Nature. 1982 Jul 29;298(5873):478–481. doi: 10.1038/298478a0. [DOI] [PubMed] [Google Scholar]
  16. Mohandas N., Rossi M. E., Clark M. R. Association between morphologic distortion of sickle cells and deoxygenation-induced cation permeability increase. Blood. 1986 Aug;68(2):450–454. [PubMed] [Google Scholar]
  17. Ortiz O. E., Lew V. L., Bookchin R. M. Calcium accumulated by sickle cell anemia red cells does not affect their potassium (86Rb+) flux components. Blood. 1986 Mar;67(3):710–715. [PubMed] [Google Scholar]
  18. Ortiz O. E., Lew V. L., Bookchin R. M. Deoxygenation permeabilizes sickle cell anaemia red cells to magnesium and reverses its gradient in the dense cells. J Physiol. 1990 Aug;427:211–226. doi: 10.1113/jphysiol.1990.sp018168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Palek J., Thomae M., Ozog D. Red cell calcium content and transmembrane calcium movements in sickle cell anemia. J Lab Clin Med. 1977 Jun;89(6):1365–1374. [PubMed] [Google Scholar]
  20. Payne J. A., Lytle C., McManus T. J. Foreign anion substitution for chloride in human red blood cells: effect on ionic and osmotic equilibria. Am J Physiol. 1990 Nov;259(5 Pt 1):C819–C827. doi: 10.1152/ajpcell.1990.259.5.C819. [DOI] [PubMed] [Google Scholar]
  21. Raftos J. E., Bookchin R. M., Lew V. L. Distribution of chloride permeabilities in normal human red cells. J Physiol. 1996 Mar 15;491(Pt 3):773–777. doi: 10.1113/jphysiol.1996.sp021256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. TOSTESON D. C., CARLSEN E., DUNHAM E. T. The effects of sickling on ion transport. I. Effect of sickling on potassium transport. J Gen Physiol. 1955 Sep 20;39(1):31–53. doi: 10.1085/jgp.39.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TOSTESON D. C. The effects of sickling on ion transport. II. The effect of sickling on sodium and cesium transport. J Gen Physiol. 1955 Sep 20;39(1):55–67. doi: 10.1085/jgp.39.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tiffert T., Etzion Z., Bookchin R. M., Lew V. L. Effects of deoxygenation on active and passive Ca2+ transport and cytoplasmic Ca2+ buffering in normal human red cells. J Physiol. 1993 May;464:529–544. doi: 10.1113/jphysiol.1993.sp019649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tiffert T., Spivak J. L., Lew V. L. Magnitude of calcium influx required to induce dehydration of normal human red cells. Biochim Biophys Acta. 1988 Aug 18;943(2):157–165. doi: 10.1016/0005-2736(88)90547-0. [DOI] [PubMed] [Google Scholar]
  26. Walder J. A., Chatterjee R., Steck T. L., Low P. S., Musso G. F., Kaiser E. T., Rogers P. H., Arnone A. The interaction of hemoglobin with the cytoplasmic domain of band 3 of the human erythrocyte membrane. J Biol Chem. 1984 Aug 25;259(16):10238–10246. [PubMed] [Google Scholar]
  27. Wieth J. O. Effects of bicarbonate and thiocyanate on fluxes of Na and K, and on glucose metabolism of actively transporting human red cells. Acta Physiol Scand. 1969 Mar;75(3):313–329. doi: 10.1111/j.1748-1716.1969.tb04384.x. [DOI] [PubMed] [Google Scholar]

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