Skip to main content
The Journal of General Physiology logoLink to The Journal of General Physiology
. 1987 Feb 1;89(2):185–213. doi: 10.1085/jgp.89.2.185

Cytoplasmic [Ca2+] and intracellular pH in lymphocytes. Role of membrane potential and volume-activated Na+/H+ exchange

PMCID: PMC2215895  PMID: 3559512

Abstract

The effect of elevating cytoplasmic Ca2+ [( Ca2+]i) on the intracellular pH (pHi) of thymic lymphocytes was investigated. In Na+- containing media, treatment of the cells with ionomycin, a divalent cation ionophore, induced a moderate cytoplasmic alkalinization. In the presence of amiloride or in Na+-free media, an acidification was observed. This acidification is at least partly due to H+ (equivalent) uptake in response to membrane hyperpolarization since: it was enhanced by pretreatment with conductive protonophores, it could be mimicked by valinomycin, and it was decreased by depolarization with K+ or gramicidin. In addition, activation of metabolic H+ production also contributes to the acidification. The alkalinization is due to Na+/H+ exchange inasmuch as it is Na+ dependent, amiloride sensitive, and accompanied by H+ efflux and net Na+ gain. A shift in the pHi dependence underlies the activation of the antiport. The effect of [Ca2+]i on Na+/H+ exchange was not associated with redistribution of protein kinase C and was also observed in cells previously depleted of this enzyme. Treatment with ionomycin induced significant cell shrinking. Prevention of shrinking largely eliminated the activation of the antiport. Moreover, a comparable shrinking produced by hypertonic media also activated the antiport. It is concluded that stimulation of Na+/H+ exchange by elevation of [Ca2+]i is due, at least in part, to cell shrinking and does not require stimulation of protein kinase C.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aronson P. S. Kinetic properties of the plasma membrane Na+-H+ exchanger. Annu Rev Physiol. 1985;47:545–560. doi: 10.1146/annurev.ph.47.030185.002553. [DOI] [PubMed] [Google Scholar]
  2. Aronson P. S. Mechanisms of active H+ secretion in the proximal tubule. Am J Physiol. 1983 Dec;245(6):F647–F659. doi: 10.1152/ajprenal.1983.245.6.F647. [DOI] [PubMed] [Google Scholar]
  3. Aronson P. S., Nee J., Suhm M. A. Modifier role of internal H+ in activating the Na+-H+ exchanger in renal microvillus membrane vesicles. Nature. 1982 Sep 9;299(5879):161–163. doi: 10.1038/299161a0. [DOI] [PubMed] [Google Scholar]
  4. Burns C. P., Rozengurt E. Serum, platelet-derived growth factor, vasopressin and phorbol esters increase intracellular pH in Swiss 3T3 cells. Biochem Biophys Res Commun. 1983 Nov 15;116(3):931–938. doi: 10.1016/s0006-291x(83)80231-9. [DOI] [PubMed] [Google Scholar]
  5. Busa W. B., Nuccitelli R. Metabolic regulation via intracellular pH. Am J Physiol. 1984 Apr;246(4 Pt 2):R409–R438. doi: 10.1152/ajpregu.1984.246.4.R409. [DOI] [PubMed] [Google Scholar]
  6. Cala P. M. Volume regulation by Amphiuma red blood cells: strategies for identifying alkali metal/H+ transport. Fed Proc. 1985 Jun;44(9):2500–2507. [PubMed] [Google Scholar]
  7. Collins M. K., Rozengurt E. Homologous and heterologous mitogenic desensitization of Swiss 3T3 cells to phorbol esters and vasopressin: role of receptor and postreceptor steps. J Cell Physiol. 1984 Feb;118(2):133–142. doi: 10.1002/jcp.1041180205. [DOI] [PubMed] [Google Scholar]
  8. Frelin C., Vigne P., Lazdunski M. The amiloride-sensitive Na+/H+ antiport in 3T3 fibroblasts. J Biol Chem. 1983 May 25;258(10):6272–6276. [PubMed] [Google Scholar]
  9. Grinstein S., Dupre A., Rothstein A. Volume regulation by human lymphocytes. Role of calcium. J Gen Physiol. 1982 May;79(5):849–868. doi: 10.1085/jgp.79.5.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grinstein S., Goetz-Smith J. D., Stewart D., Beresford B. J., Mellors A. Protein phosphorylation during activation of Na+/H+ exchange by phorbol esters and by osmotic shrinking. Possible relation to cell pH and volume regulation. J Biol Chem. 1986 Jun 15;261(17):8009–8016. [PubMed] [Google Scholar]
  11. Grinstein S., Mack E., Mills G. B. Osmotic activation of the Na+/H+ antiport in protein kinase C-depleted lymphocytes. Biochem Biophys Res Commun. 1986 Jan 14;134(1):8–13. doi: 10.1016/0006-291x(86)90519-x. [DOI] [PubMed] [Google Scholar]
  12. Grinstein S., Rothstein A. Mechanisms of regulation of the Na+/H+ exchanger. J Membr Biol. 1986;90(1):1–12. doi: 10.1007/BF01869680. [DOI] [PubMed] [Google Scholar]
  13. Guy G. R., Gordon J., Walker L., Michell R. H., Brown G. Redistribution of protein kinase C during mitogenesis of human B lymphocytes. Biochem Biophys Res Commun. 1986 Feb 26;135(1):146–153. doi: 10.1016/0006-291x(86)90954-x. [DOI] [PubMed] [Google Scholar]
  14. Hesketh T. R., Moore J. P., Morris J. D., Taylor M. V., Rogers J., Smith G. A., Metcalfe J. C. A common sequence of calcium and pH signals in the mitogenic stimulation of eukaryotic cells. Nature. 1985 Feb 7;313(6002):481–484. doi: 10.1038/313481a0. [DOI] [PubMed] [Google Scholar]
  15. Hoffmann E. K. Role of separate K+ and Cl- channels and of Na+/Cl- cotransport in volume regulation in Ehrlich cells. Fed Proc. 1985 Jun;44(9):2513–2519. [PubMed] [Google Scholar]
  16. Hoffmann E. K., Simonsen L. O., Lambert I. H. Volume-induced increase of K+ and Cl- permeabilities in Ehrlich ascites tumor cells. Role of internal Ca2+. J Membr Biol. 1984;78(3):211–222. doi: 10.1007/BF01925969. [DOI] [PubMed] [Google Scholar]
  17. Kauffman R. F., Taylor R. W., Pfeiffer D. R. Cation transport and specificity of ionomycin. Comparison with ionophore A23187 in rat liver mitochondria. J Biol Chem. 1980 Apr 10;255(7):2735–2739. [PubMed] [Google Scholar]
  18. Kraft A. S., Anderson W. B. Phorbol esters increase the amount of Ca2+, phospholipid-dependent protein kinase associated with plasma membrane. Nature. 1983 Feb 17;301(5901):621–623. doi: 10.1038/301621a0. [DOI] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Mahnensmith R. L., Aronson P. S. The plasma membrane sodium-hydrogen exchanger and its role in physiological and pathophysiological processes. Circ Res. 1985 Jun;56(6):773–788. doi: 10.1161/01.res.56.6.773. [DOI] [PubMed] [Google Scholar]
  21. Melloni E., Pontremoli S., Michetti M., Sacco O., Sparatore B., Horecker B. L. The involvement of calpain in the activation of protein kinase C in neutrophils stimulated by phorbol myristic acid. J Biol Chem. 1986 Mar 25;261(9):4101–4105. [PubMed] [Google Scholar]
  22. Molski T. F., Naccache P. H., Volpi M., Wolpert L. M., Sha'afi R. I. Specific modulation of the intracellular pH of rabbit neutrophils by chemotactic factors. Biochem Biophys Res Commun. 1980 May 30;94(2):508–514. doi: 10.1016/0006-291x(80)91260-7. [DOI] [PubMed] [Google Scholar]
  23. Moolenaar W. H., Mummery C. L., van der Saag P. T., de Laat S. W. Rapid ionic events and the initiation of growth in serum-stimulated neuroblastoma cells. Cell. 1981 Mar;23(3):789–798. doi: 10.1016/0092-8674(81)90443-8. [DOI] [PubMed] [Google Scholar]
  24. Moolenaar W. H., Tertoolen L. G., de Laat S. W. Phorbol ester and diacylglycerol mimic growth factors in raising cytoplasmic pH. Nature. 1984 Nov 22;312(5992):371–374. doi: 10.1038/312371a0. [DOI] [PubMed] [Google Scholar]
  25. Moolenaar W. H., Tsien R. Y., van der Saag P. T., de Laat S. W. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. doi: 10.1038/304645a0. [DOI] [PubMed] [Google Scholar]
  26. Moore R. D. Stimulation of Na:H exchange by insulin. Biophys J. 1981 Feb;33(2):203–210. doi: 10.1016/S0006-3495(81)84881-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Muldoon L. L., Dinerstein R. J., Villereal M. L. Intracellular pH in human fibroblasts: effect of mitogens, A23187, and phospholipase activation. Am J Physiol. 1985 Jul;249(1 Pt 1):C140–C148. doi: 10.1152/ajpcell.1985.249.1.C140. [DOI] [PubMed] [Google Scholar]
  28. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  29. Owen N. E., Villereal M. L. Effect of the intracellular Ca+2 antagonist TMB-8 on serum-stimulated Na+ influx in human fibroblasts. Biochem Biophys Res Commun. 1982 Dec 15;109(3):762–768. doi: 10.1016/0006-291x(82)92005-8. [DOI] [PubMed] [Google Scholar]
  30. Rink R. J., Sanchez A., Grinstein S., Rothstein A. Volume restoration in osmotically swollen lymphocytes does not involve changes in free Ca2+ concentration. Biochim Biophys Acta. 1983 Jul 14;762(4):593–596. doi: 10.1016/0167-4889(83)90064-2. [DOI] [PubMed] [Google Scholar]
  31. Rink T. J., Montecucco C., Hesketh T. R., Tsien R. Y. Lymphocyte membrane potential assessed with fluorescent probes. Biochim Biophys Acta. 1980;595(1):15–30. doi: 10.1016/0005-2736(80)90243-6. [DOI] [PubMed] [Google Scholar]
  32. Rink T. J., Tsien R. Y., Pozzan T. Cytoplasmic pH and free Mg2+ in lymphocytes. J Cell Biol. 1982 Oct;95(1):189–196. doi: 10.1083/jcb.95.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rodriguez-Pena A., Rozengurt E. Disappearance of Ca2+-sensitive, phospholipid-dependent protein kinase activity in phorbol ester-treated 3T3 cells. Biochem Biophys Res Commun. 1984 May 16;120(3):1053–1059. doi: 10.1016/s0006-291x(84)80213-2. [DOI] [PubMed] [Google Scholar]
  34. Rothenberg P., Glaser L., Schlesinger P., Cassel D. Activation of Na+/H+ exchange by epidermal growth factor elevates intracellular pH in A431 cells. J Biol Chem. 1983 Oct 25;258(20):12644–12653. [PubMed] [Google Scholar]
  35. Schuldiner S., Rozengurt E. Na+/H+ antiport in Swiss 3T3 cells: mitogenic stimulation leads to cytoplasmic alkalinization. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7778–7782. doi: 10.1073/pnas.79.24.7778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Siffert W., Fox G., Mückenhoff K., Scheid P. Thrombin stimulates Na+-H+ exchange across the human platelet plasma membrane. FEBS Lett. 1984 Jul 9;172(2):272–274. doi: 10.1016/0014-5793(84)81139-4. [DOI] [PubMed] [Google Scholar]
  37. Thomas J. A., Buchsbaum R. N., Zimniak A., Racker E. Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. Biochemistry. 1979 May 29;18(11):2210–2218. doi: 10.1021/bi00578a012. [DOI] [PubMed] [Google Scholar]
  38. Thomas R. C. Experimental displacement of intracellular pH and the mechanism of its subsequent recovery. J Physiol. 1984 Sep;354:3P–22P. doi: 10.1113/jphysiol.1984.sp015397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vara F., Rozengurt E. Stimulation of Na+/H+ antiport activity by epidermal growth factor and insulin occurs without activation of protein kinase C. Biochem Biophys Res Commun. 1985 Jul 31;130(2):646–653. doi: 10.1016/0006-291x(85)90466-8. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of General Physiology are provided here courtesy of The Rockefeller University Press

RESOURCES