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. 1996 Oct 1;15(19):5177–5183.

A slow sustained increase in cytosolic Ca2+ levels mediates stalk gene induction by differentiation inducing factor in Dictyostelium.

P Schaap 1, T Nebl 1, P R Fisher 1
PMCID: PMC452261  PMID: 8895562

Abstract

During Dictyostelium stalk cell differentiation, cells vacuolate, synthesize a cellulose cell wall and die. This process of programmed cell death is accompanied by expression of the prestalk gene ecmB and induced by the differentiation inducing factor DIF. Using cell lines expressing the recombinant Ca2+-sensitive photoprotein apoaequorin, we found that 100 nM DIF increases cytosolic Ca2+ ([Ca2+]i) levels from approximately 50 to 150 nM over a period of 8 h. The Ca2+-ATPase inhibitor 2,5-di(tert-butyl)-1,4-hydroquinone (BHQ) induced a similar increase in [Ca2+]i levels and induced expression of the prestalk gene ecmB to the same level as DIF. The [Ca2+]i increases induced by DIF and BHQ showed similar kinetics and preceded ecmB gene expression by approximately 1-2 h. The Ca2+ chelator 1,2-bis(o-aminophenoxy)-ethane-N,N,N'N'-tetra-acetic acid (BAPTA) efficiently inhibited the BHQ-induced [Ca2+]i increase and blocked DIF-induced expression of the ecmB gene. These data indicate that the effects of DIF on stalk gene expression are mediated by a sustained increase in [Ca2-]i. Sustained [Ca2+]i elevation mediates many forms of programmed cell death in vertebrates. The Dictyostelium system may be the earliest example of how this mechanism developed during early eukaryote evolution.

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Selected References

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  1. Berks M., Kay R. R. Combinatorial control of cell differentiation by cAMP and DIF-1 during development of Dictyostelium discoideum. Development. 1990 Nov;110(3):977–984. doi: 10.1242/dev.110.3.977. [DOI] [PubMed] [Google Scholar]
  2. Ceccarelli A., Mahbubani H., Williams J. G. Positively and negatively acting signals regulating stalk cell and anterior-like cell differentiation in Dictyostelium. Cell. 1991 Jun 14;65(6):983–989. doi: 10.1016/0092-8674(91)90550-i. [DOI] [PubMed] [Google Scholar]
  3. Chow S. C., Kass G. E., McCabe M. J., Jr, Orrenius S. Tributyltin increases cytosolic free Ca2+ concentration in thymocytes by mobilizing intracellular Ca2+, activating a Ca2+ entry pathway, and inhibiting Ca2+ efflux. Arch Biochem Biophys. 1992 Oct;298(1):143–149. doi: 10.1016/0003-9861(92)90105-6. [DOI] [PubMed] [Google Scholar]
  4. Clapham D. E. Calcium signaling. Cell. 1995 Jan 27;80(2):259–268. doi: 10.1016/0092-8674(95)90408-5. [DOI] [PubMed] [Google Scholar]
  5. Cobbold P. H., Rink T. J. Fluorescence and bioluminescence measurement of cytoplasmic free calcium. Biochem J. 1987 Dec 1;248(2):313–328. doi: 10.1042/bj2480313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cornillon S., Foa C., Davoust J., Buonavista N., Gross J. D., Golstein P. Programmed cell death in Dictyostelium. J Cell Sci. 1994 Oct;107(Pt 10):2691–2704. doi: 10.1242/jcs.107.10.2691. [DOI] [PubMed] [Google Scholar]
  7. Coukell M. B., Cameron A. M. Effects of suboptimal levels of extracellular calcium on the regulation of the cyclic AMP phosphodiesterase-inhibitor system and membrane differentiation in Dictyostelium discoideum. J Cell Sci. 1988 Aug;90(Pt 4):691–700. doi: 10.1242/jcs.90.4.691. [DOI] [PubMed] [Google Scholar]
  8. Cubitt A. B., Firtel R. A., Fischer G., Jaffe L. F., Miller A. L. Patterns of free calcium in multicellular stages of Dictyostelium expressing jellyfish apoaequorin. Development. 1995 Aug;121(8):2291–2301. doi: 10.1242/dev.121.8.2291. [DOI] [PubMed] [Google Scholar]
  9. Devreotes P. N. G protein-linked signaling pathways control the developmental program of Dictyostelium. Neuron. 1994 Feb;12(2):235–241. doi: 10.1016/0896-6273(94)90267-4. [DOI] [PubMed] [Google Scholar]
  10. Dingermann T., Reindl N., Werner H., Hildebrandt M., Nellen W., Harwood A., Williams J., Nerke K. Optimization and in situ detection of Escherichia coli beta-galactosidase gene expression in Dictyostelium discoideum. Gene. 1989 Dec 28;85(2):353–362. doi: 10.1016/0378-1119(89)90428-9. [DOI] [PubMed] [Google Scholar]
  11. Dowd D. R. Calcium regulation of apoptosis. Adv Second Messenger Phosphoprotein Res. 1995;30:255–280. doi: 10.1016/s1040-7952(05)80010-2. [DOI] [PubMed] [Google Scholar]
  12. Europe-Finner G. N., Newell P. C. Inositol 1,4,5-triphosphate induces calcium release from a non- mitochondrial pool in amoebae of Dictyostelium. Biochim Biophys Acta. 1986 Aug 1;887(3):335–340. doi: 10.1016/0167-4889(86)90163-1. [DOI] [PubMed] [Google Scholar]
  13. Firtel R. A. Integration of signaling information in controlling cell-fate decisions in Dictyostelium. Genes Dev. 1995 Jun 15;9(12):1427–1444. doi: 10.1101/gad.9.12.1427. [DOI] [PubMed] [Google Scholar]
  14. Ghosh T. K., Bian J. H., Short A. D., Rybak S. L., Gill D. L. Persistent intracellular calcium pool depletion by thapsigargin and its influence on cell growth. J Biol Chem. 1991 Dec 25;266(36):24690–24697. [PubMed] [Google Scholar]
  15. Heuser J., Zhu Q., Clarke M. Proton pumps populate the contractile vacuoles of Dictyostelium amoebae. J Cell Biol. 1993 Jun;121(6):1311–1327. doi: 10.1083/jcb.121.6.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hopper N. A., Anjard C., Reymond C. D., Williams J. G. Induction of terminal differentiation of Dictyostelium by cAMP-dependent protein kinase and opposing effects of intracellulr and extracellular cAMP on stalk cell differentiation. Development. 1993 Sep;119(1):147–154. doi: 10.1242/dev.119.1.147. [DOI] [PubMed] [Google Scholar]
  17. Inesi G., Sagara Y. Thapsigargin, a high affinity and global inhibitor of intracellular Ca2+ transport ATPases. Arch Biochem Biophys. 1992 Nov 1;298(2):313–317. doi: 10.1016/0003-9861(92)90416-t. [DOI] [PubMed] [Google Scholar]
  18. Insall R., Kay R. R. A specific DIF binding protein in Dictyostelium. EMBO J. 1990 Oct;9(10):3323–3328. doi: 10.1002/j.1460-2075.1990.tb07532.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Insall R., Nayler O., Kay R. R. DIF-1 induces its own breakdown in Dictyostelium. EMBO J. 1992 Aug;11(8):2849–2854. doi: 10.1002/j.1460-2075.1992.tb05352.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jackson T. R., Patterson S. I., Thastrup O., Hanley M. R. A novel tumour promoter, thapsigargin, transiently increases cytoplasmic free Ca2+ without generation of inositol phosphates in NG115-401L neuronal cells. Biochem J. 1988 Jul 1;253(1):81–86. doi: 10.1042/bj2530081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kaiser N., Edelman I. S. Calcium dependence of glucocorticoid-induced lymphocytolysis. Proc Natl Acad Sci U S A. 1977 Feb;74(2):638–642. doi: 10.1073/pnas.74.2.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kay R. R., Jermyn K. A. A possible morphogen controlling differentiation in Dictyostelium. Nature. 1983 May 19;303(5914):242–244. doi: 10.1038/303242a0. [DOI] [PubMed] [Google Scholar]
  23. Knight M. R., Campbell A. K., Smith S. M., Trewavas A. J. Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. Nature. 1991 Aug 8;352(6335):524–526. doi: 10.1038/352524a0. [DOI] [PubMed] [Google Scholar]
  24. Kubohara Y., Okamoto K. Cytoplasmic Ca2+ and H+ concentrations determine cell fate in Dictyostelium discoideum. FASEB J. 1994 Aug;8(11):869–874. doi: 10.1096/fasebj.8.11.8070636. [DOI] [PubMed] [Google Scholar]
  25. Kubohara Y., Saito Y., Tatemoto K. Differentiation-inducing factor of D. discoideum raises intracellular calcium concentration and suppresses cell growth in rat pancreatic AR42J cells. FEBS Lett. 1995 Feb 13;359(2-3):119–122. doi: 10.1016/0014-5793(95)00022-2. [DOI] [PubMed] [Google Scholar]
  26. Lam M., Dubyak G., Distelhorst C. W. Effect of glucocorticosteroid treatment on intracellular calcium homeostasis in mouse lymphoma cells. Mol Endocrinol. 1993 May;7(5):686–693. doi: 10.1210/mend.7.5.8316252. [DOI] [PubMed] [Google Scholar]
  27. Maeda Y., Maeda M. The calcium content of the cellular slime mold, Dictyostelium discoideum, during development and differentiation. Exp Cell Res. 1973 Nov;82(1):125–130. doi: 10.1016/0014-4827(73)90253-x. [DOI] [PubMed] [Google Scholar]
  28. McConkey D. J., Nicotera P., Hartzell P., Bellomo G., Wyllie A. H., Orrenius S. Glucocorticoids activate a suicide process in thymocytes through an elevation of cytosolic Ca2+ concentration. Arch Biochem Biophys. 1989 Feb 15;269(1):365–370. doi: 10.1016/0003-9861(89)90119-7. [DOI] [PubMed] [Google Scholar]
  29. Milne J. L., Coukell M. B. Identification of a high-affinity Ca2+ pump associated with endocytotic vesicles in Dictyostelium discoideum. Exp Cell Res. 1989 Nov;185(1):21–32. doi: 10.1016/0014-4827(89)90033-5. [DOI] [PubMed] [Google Scholar]
  30. Nellen W., Datta S., Reymond C., Sivertsen A., Mann S., Crowley T., Firtel R. A. Molecular biology in Dictyostelium: tools and applications. Methods Cell Biol. 1987;28:67–100. doi: 10.1016/s0091-679x(08)61637-4. [DOI] [PubMed] [Google Scholar]
  31. Nolta K. V., Steck T. L. Isolation and initial characterization of the bipartite contractile vacuole complex from Dictyostelium discoideum. J Biol Chem. 1994 Jan 21;269(3):2225–2233. [PubMed] [Google Scholar]
  32. Oldershaw K. A., Taylor C. W. 2,5-Di-(tert-butyl)-1,4-benzohydroquinone mobilizes inositol 1,4,5-trisphosphate-sensitive and -insensitive Ca2+ stores. FEBS Lett. 1990 Nov 12;274(1-2):214–216. doi: 10.1016/0014-5793(90)81366-v. [DOI] [PubMed] [Google Scholar]
  33. Rooney E. K., Gross J. D. ATP-driven Ca2+/H+ antiport in acid vesicles from Dictyostelium. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8025–8029. doi: 10.1073/pnas.89.17.8025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rooney E. K., Gross J. D., Satre M. Characterisation of an intracellular Ca2+ pump in Dictyostelium. Cell Calcium. 1994 Dec;16(6):509–522. doi: 10.1016/0143-4160(94)90081-7. [DOI] [PubMed] [Google Scholar]
  35. Saran S., Nakao H., Tasaka M., Iida H., Tsuji F. I., Nanjundiah V., Takeuchi I. Intracellular free calcium level and its response to cAMP stimulation in developing Dictyostelium cells transformed with jellyfish apoaequorin cDNA. FEBS Lett. 1994 Jan 3;337(1):43–47. doi: 10.1016/0014-5793(94)80626-8. [DOI] [PubMed] [Google Scholar]
  36. Shimomura O., Musicki B., Kishi Y. Semi-synthetic aequorins with improved sensitivity to Ca2+ ions. Biochem J. 1989 Aug 1;261(3):913–920. doi: 10.1042/bj2610913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tirlapur U. K., Gross J., Nanjundiah V. Spatial variation of sequestered calcium in the multicellular stage of Dictyostelium discoideum as assayed by chlortetracycline fluorescence. Differentiation. 1991 Dec;48(3):137–146. doi: 10.1111/j.1432-0436.1991.tb00252.x. [DOI] [PubMed] [Google Scholar]
  38. Trump B. F., Berezesky I. K. The role of cytosolic Ca2+ in cell injury, necrosis and apoptosis. Curr Opin Cell Biol. 1992 Apr;4(2):227–232. doi: 10.1016/0955-0674(92)90037-d. [DOI] [PubMed] [Google Scholar]
  39. Wang M., Roelfsema J. H., Williams J. G., Schaap P. Cytoplasmic acidification facilitates but does not mediate DIF-induced prestalk gene expression in Dictyostelium discoideum. Dev Biol. 1990 Jul;140(1):182–188. doi: 10.1016/0012-1606(90)90065-q. [DOI] [PubMed] [Google Scholar]
  40. Williams J. G., Ceccarelli A., McRobbie S., Mahbubani H., Kay R. R., Early A., Berks M., Jermyn K. A. Direct induction of Dictyostelium prestalk gene expression by DIF provides evidence that DIF is a morphogen. Cell. 1987 Apr 24;49(2):185–192. doi: 10.1016/0092-8674(87)90559-9. [DOI] [PubMed] [Google Scholar]
  41. Witke W., Nellen W., Noegel A. Homologous recombination in the Dictyostelium alpha-actinin gene leads to an altered mRNA and lack of the protein. EMBO J. 1987 Dec 20;6(13):4143–4148. doi: 10.1002/j.1460-2075.1987.tb02760.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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