1887

Abstract

This study presents morphological and biochemical evidence of programmed cell death (PCD) in induced by exposure of trophozoites to the aminoglycoside antibiotic G418. Morphological characteristics of PCD, including cell shrinkage, reduced cellular volume, nuclear condensation, DNA fragmentation and vacuolization were observed, with preservation of trophozoite membrane integrity. PCD is orchestrated biochemically by alterations in intracellular ion fluxes. In G418-treated trophozoites, overproduction of reactive oxygen species (ROS), decreased intracellular K, increased cytosolic calcium, and decreased intracellular pH levels were observed. However, externalization of phosphatidylserine was not detected. These results suggest that amoebae can undergo PCD under stress conditions, and that this PCD shares several properties with PCD reported in mammals and in a variety of unicellular organisms.

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2007-11-01
2019-10-17
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References

  1. Al-Olayan, E. M., Williams, G. T. & Hurd, H. ( 2002; ). Apoptosis in the malaria protozoan, Plasmodium berghei: a possible mechanism for limiting intensity of infection in the mosquito. Int J Parasitol 32, 1133–1143.[CrossRef]
    [Google Scholar]
  2. Aley, S. B., Scott, W. A. & Cohn, Z. A. ( 1980; ). Plasma membrane of Entamoeba histolytica. J Exp Med 152, 391–404.[CrossRef]
    [Google Scholar]
  3. Ameisen, J. C., Estaquier, J. & Idziorek, T. ( 1994; ). From AIDS to parasite infection: pathogen-mediated subversion of programmed cell death as a mechanism for immune dysregulation. Immunol Rev 142, 9–51.[CrossRef]
    [Google Scholar]
  4. Barbieri, D., Troiano, L., Grassilli, E., Agnesini, C., Cristofalo, E. A., Monti, D., Capri, M., Cossarizza, A. & Franceschi, C. ( 1992; ). Inhibition of apoptosis by zinc: a reappraisal. Biochem Biophys Res Commun 187, 1256–1261.[CrossRef]
    [Google Scholar]
  5. Berninghausen, O. & Leippe, M. ( 1997; ). Necrosis versus apoptosis as the mechanism of target cell death induced by Entamoeba histolytica. Infect Immun 65, 3615–3621.
    [Google Scholar]
  6. Billaut-Mulot, O., Fernandez-Gomez, R., Loyens, M. & Ouaissis, A. ( 1996; ). Trypanosoma cruzi elongation factor 1-α: nuclear localization in parasites undergoing apoptosis. Gene 174, 19–26.[CrossRef]
    [Google Scholar]
  7. Bortner, C. D., Hughes, F. M., Jr & Cidlowski, J. A. ( 1997; ). A primary role for K+ and Na+ efflux in the activation of apoptosis. J Biol Chem 272, 32436–32442.[CrossRef]
    [Google Scholar]
  8. Bruchhaus, I., Loftus, B. J., Hall, N. & Tannich, E. ( 2003; ). The intestinal protozoan parasite Entamoeba histolytica contains 20 cysteine protease genes, of which only a small subset is expressed during in vitro cultivation. Eukaryot Cell 2, 501–509.[CrossRef]
    [Google Scholar]
  9. Chen, G., Branton, P. E. & Shore, G. C. ( 1995; ). Induction of p53-independent apoptosis by hygromycin B: suppression by Bcl-2 and adenovirus E1B 19-kDa protein. Exp Cell Res 221, 55–59.[CrossRef]
    [Google Scholar]
  10. Christensen, S. T., Wheatley, D. N., Rasmussen, M. I. & Rasmussen, L. ( 1995; ). Mechanisms controlling death, survival and proliferation in a model unicellular eukaryote Tetrahymena thermophila. Cell Death Differ 2, 301–308.
    [Google Scholar]
  11. Clarke, P. G. H. ( 1990; ). Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol (Berl) 181, 195–213.
    [Google Scholar]
  12. Collins, R. J., Harmon, B. V., Gobe, G. C. & Kerr, J. F. R. ( 1992; ). Internucleosomal DNA cleavage should not be the sole criterion for identifying apoptosis. Int J Radiat Biol 61, 451–453.[CrossRef]
    [Google Scholar]
  13. Cornillon, S., Foa, C., Davoust, J., Buonavista, N. & Gross, J. D. ( 1994; ). Programmed cell death in Dictyostelium. J Cell Sci 107, 2691–2704.
    [Google Scholar]
  14. Dale, C., Welburn, S. C., Maudlin, I. & Milligan, P. J. M. ( 1995; ). The kinetics of maturation of trypanosome infections in tsetse. Parasitology 111, 187–191.[CrossRef]
    [Google Scholar]
  15. Demaurex, N., Frieden, M. & Arnaudeau, S. ( 2003; ). ER calcium and ER chaperones: new players in apoptosis? In Calreticulin, 2nd edn, pp. 134–142. Edited by P. Eggleton & M. Michalak. Austin, TX: Eurekah.
  16. Diamond, L. S., Harlow, D. R. & Cunnick, C. C. ( 1978; ). A new medium for axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 72, 431–432.[CrossRef]
    [Google Scholar]
  17. Didenko, V. V., Ngo, H. & Baskin, D. S. ( 2003; ). Early necrotic DNA degradation: presence of blunt-ended DNA breaks, 3′ and 5′ overhangs in apoptosis, but only 5′ overhangs in early necrosis. Am J Pathol 162, 1571–1578.[CrossRef]
    [Google Scholar]
  18. Espinosa-Cantellano, M. & Martínez-Palomo, A. ( 2000; ). Pathogenesis of intestinal amebiasis: from molecules to disease. Clin Microbiol Rev 13, 318–331.[CrossRef]
    [Google Scholar]
  19. Facompre, M., Goossens, J. F. & Bailly, C. ( 2001; ). Apoptotic response of HL-60 human leukemia cells to the antitumor drug NB-506, a glycosylated indolocarbazole inhibitor of topoisomerase 1. Biochem Pharmacol 61, 299–310.[CrossRef]
    [Google Scholar]
  20. Falcieri, E., Martelli, A. M., Bareggi, R., Cataldi, A. & Cocco, L. ( 1993; ). The protein kinase inhibitor staurosporine induces morphological changes typical of apoptosis in MOLT-4 cells without concomitant DNA fragmentation. Biochem Biophys Res Commun 193, 19–25.[CrossRef]
    [Google Scholar]
  21. Freire-de-Lima, C. G., Nascimento, D. O., Soares, M. B. P., Bozza, P. T., Castro-Faria-Neto, H. C., de Mello, F. G., DosReisand, G. A. & Lopes, M. F. ( 2000; ). Uptake of apoptotic cells drives the growth of a pathogenic trypanosome in macrophages. Nature 403, 199–203.[CrossRef]
    [Google Scholar]
  22. Gatti, R., Belletti, S., Orlandini, G., Bussolati, O., Dall'Asta, V. & Gazzola, G. C. ( 1998; ). Comparison of Annexin V and Calcein-AM as early vital markers of apoptosis in adherent cells by confocal laser microscopy. J Histochem Cytochem 46, 895–900.[CrossRef]
    [Google Scholar]
  23. Gómez, C., Pérez, D. G., López-Bayghens, E. & Orozco, E. ( 1998; ). Transcriptional analysis of the EhPgp1 promoter of Entamoeba histolytica multidrug-resistant mutant. J Biol Chem 273, 7277–7284.[CrossRef]
    [Google Scholar]
  24. Grynkiewicz, G., Poenie, M. & Tsien, R. Y. ( 1985; ). A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260, 3440–3450.
    [Google Scholar]
  25. Halliwell, B. & Gutteridge, J. M. C. ( 1989; ). Free Radicals in Biology and Medicine, 2nd edn. Oxford: Clarendon Press.
  26. Hawley, T. S. & Hawley, R. G. ( 2004; ). Methods in Molecular Biology: Flow Cytometry Protocols, 2nd edn. Totowa, NJ: Humana Press.
  27. Hesse, F., Selzer, P. M., Muhlstadt, K. & Duszenko, M. ( 1995; ). A novel cultivation technique for long-term maintenance of bloodstream form trypanosomes in vitro. Mol Biochem Parasitol 70, 157–166.[CrossRef]
    [Google Scholar]
  28. Hirata, H., Hibasami, H., Yoshida, T., Morita, A., Ohkaya, S., Matsumoto, M., Sasaki, H. & Uchida, A. ( 1998; ). Differentiation and apoptosis without DNA fragmentation in cultured Schwann cells derived from wallerian-degenerated nerve. Apoptosis 3, 353–360.[CrossRef]
    [Google Scholar]
  29. Howell, D. M. & Martz, E. ( 1987; ). The degree of CTL-induced DNA solubilization is not determined by the human vs mouse origin of the target cell. J Immunol 138, 3695–3698.
    [Google Scholar]
  30. Huppertz, B., Frank, H. G. & Kaufmann, P. ( 1999; ). The apoptosis cascade – morphological and inmunohistochemical methods for its visualization. Anat Embryol (Berl) 200, 1–18.[CrossRef]
    [Google Scholar]
  31. Huston, C. D., Boettner, D. R., Miller-Sims, V. & Petri, W. A. J. ( 2003; ). Apoptotic killing and phagocytosis of host cells by the parasite Entamoeba histolytica. Infect Immun 71, 964–972.[CrossRef]
    [Google Scholar]
  32. Jacobson, M. D., Weil, M. & Raff, M. C. ( 1997; ). Programmed cell death in animal development. Cell 88, 347–354.[CrossRef]
    [Google Scholar]
  33. Jiang, S., Chow, S. C., Nicotera, P. & Orrenius, S. ( 1994; ). Intracellular Ca2+ signals activate apoptosis in thymocytes: studies using the Ca2+-ATPase inhibitor thapsigargin. Exp Cell Res 212, 84–92.[CrossRef]
    [Google Scholar]
  34. Jin, Q. H., Zhao, B. & Zhang, X. J. ( 2004; ). Cytochrome c release and endoplasmic reticulum stress are involved in caspase-dependent apoptosis induced by G418. Cell Mol Life Sci 61, 1816–1825.
    [Google Scholar]
  35. Kroemer, G. & Reed, J. C. ( 2000; ). Mitochondrial control of cell death. Nat Med 6, 513–519.[CrossRef]
    [Google Scholar]
  36. Lee, N., Bertholet, S., Debrabant, A., Muller, J., Duncan, R. & Nakhasi, H. L. ( 2002; ). Programmed cell death in the unicellular protozoan parasite Leishmania. Cell Death Differ 9, 53–64.[CrossRef]
    [Google Scholar]
  37. Lewis, K. ( 2000; ). Programmed death in bacteria. Microbiol Mol Biol Rev 64, 503–514.[CrossRef]
    [Google Scholar]
  38. Madeo, F., Frohlich, E., Ligr, M., Grey, M., Sigrist, S. J., Wolf, D. H. & Frohlich, K. U. ( 1999; ). Oxygen stress: a regulator of apoptosis in yeast. J Cell Biol 145, 757–767.[CrossRef]
    [Google Scholar]
  39. Martin, J. B., Bakker-Grunwald, T. & Klein, G. ( 1993; ). 31P-NMR analysis of Entamoeba histolytica. Occurrence of high amounts of two inositol phosphates. Eur J Biochem 214, 711–718.[CrossRef]
    [Google Scholar]
  40. Matsui, J. I., Gale, J. E. & Warchol, M. E. ( 2004; ). Critical signaling events during the aminoglycoside-induced death of sensory hair cells in vitro. J Neurobiol 61, 250–266.[CrossRef]
    [Google Scholar]
  41. Matsuyama, S., Llopis, J., Deveraux, Q. L., Tsien, R. & Reed, J. C. ( 2000; ). Changes in intramitochondrial and cytosolic pH: early events that modulate caspase activation during apoptosis. Nat Cell Biol 2, 318–325.[CrossRef]
    [Google Scholar]
  42. Mesner, P. W., Winters, T. R. & Green, S. H. ( 1992; ). Nerve growth factor withdrawal-induced cell death in neuronal PC12 cells resembles that in sympathetic neurons. J Cell Biol 119, 1669–1680.[CrossRef]
    [Google Scholar]
  43. Nasirudeen, A. M. A., Hian, Y. E., Singh, M. & Tan, K. S. W. ( 2004; ). Metronidazole induces programmed cell death in the protozoan parasite Blastocystis hominis. Microbiology 150, 33–43.[CrossRef]
    [Google Scholar]
  44. Nguewa, P. A., Fuentes, M. A., Valladares, B., Alonso, C. & Pérez, J. M. ( 2004; ). Programmed cell death in trypanosomatids: a way to maximize their biological fitness?. Trends Parasitol 20, 375–380.[CrossRef]
    [Google Scholar]
  45. Ragland, B. D., Ashley, L. S., Vaux, D. L. & Petri, W. A., Jr ( 1994; ). Entamoeba histolytica: target cells killed by trophozoites undergo DNA fragmentation which is not blocked by Bcl-2. Exp Parasitol 79, 460–467.[CrossRef]
    [Google Scholar]
  46. Ramos, E., Olivos-García, A., Nequiz, M., Saavedra, E., Tello, E., Saralegui, A., Montfort, I. & Pérez Tamayo, R. ( 2007; ). Entamoeba histolytica: apoptosis induced in vitro by nitric oxide species. Exp Parasitol 116, 257––265.[CrossRef]
    [Google Scholar]
  47. Robertson, A. M. G. & Thomson, J. N. ( 1982; ). Morphology of programmed cell death in the ventral nerve cord of Caenorhabditis elegans larvae. J Embryol Exp Morphol 67, 89–100.
    [Google Scholar]
  48. Sat, B., Hazan, R., Fisher, T., Khaner, H., Glaser, G. & Engelberg-Kulka, H. ( 2001; ). Programmed cell death in Escherichia coli: some antibiotics can trigger mazEF lethality. J Bacteriol 183, 2041–2045.[CrossRef]
    [Google Scholar]
  49. Scheuerlein, R., Treml, S., Thar, B., Tirlapur, U. K. & Hader, D. P. ( 1995; ). Evidence for UV-B-induced DNA degradation in Euglena gracilis mediated by activation of metal-dependent nucleases. J Photochem Photobiol B 31, 113–123.[CrossRef]
    [Google Scholar]
  50. Sen, N., Das, B. B., Ganguly, A., Mukherjee, T., Bandyopadhyay, S. & Majumder, H. K. ( 2004a; ). Camptothecin-induced imbalance in intracellular cation homeostasis regulates programmed cell death in unicellular hemoflagellate Leishmania donovani. J Biol Chem 279, 52366–52375.[CrossRef]
    [Google Scholar]
  51. Sen, N., Das, B. B., Ganguly, A., Mukherjee, T., Tripathi, G., Bandyopadhyay, S., Rakshit, S., Sen, T. & Majumder, H. K. ( 2004b; ). Camptothecin induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani. Cell Death Differ 11, 924–936.[CrossRef]
    [Google Scholar]
  52. Seydel, K. B. & Stanley, S. L., Jr ( 1998; ). Entamoeba histolytica induces host cell death in amebic liver abscess by a non-Fas-dependent, non-tumor necrosis factor alpha-dependent pathway of apoptosis. Infect Immun 66, 2980–2983.
    [Google Scholar]
  53. Tagliarino, C., Pink, J. J., Dubyak, G. R., Nieminen, A. L. & Boothman, D. A. ( 2001; ). Calcium is a key signaling molecule in β-lapachone-mediated cell death. J Biol Chem 276, 19150–19159.[CrossRef]
    [Google Scholar]
  54. Tandogan, B. & Ulusu, N. N. ( 2005; ). Importance of calcium. Tr J Med Sci 35, 197–201.
    [Google Scholar]
  55. Tillack, M., Biller, L., Irmer, H., Freitas, M., Gomes, M., Tannich, E. & Bruchhaus, I. ( 2007; ). The Entamoeba histolytica genome: primary structure and expression of proteolytic enzymes. BMC Genomics 8, 170 [CrossRef]
    [Google Scholar]
  56. Tomei, L. D., Shapiro, J. P. & Cope, F. O. ( 1993; ). Apoptosis in C3H/10T1/2 mouse embryonic cells: evidence for internucleosomal DNA modification in the absence of double-strand cleavage. Proc Natl Acad Sci U S A 90, 853–857.[CrossRef]
    [Google Scholar]
  57. Torres-Guerrero, H., Peattie, D. A. & Meza, I. ( 1991; ). Chromatin organization in Entamoeba histolytica. Mol Biochem Parasitol 45, 121–130.[CrossRef]
    [Google Scholar]
  58. Vardi, A., Berman-Frank, I., Rozenberg, T., Hadas, O., Kaplan, A. & Levine, A. ( 1999; ). Programmed cell death of the dinoflagellate Peridinium gatunense is mediated by CO2 limitation and oxidative stress. Curr Biol 9, 1061–1064.[CrossRef]
    [Google Scholar]
  59. Vaux, D. L., Haecker, G. & Strasser, A. ( 1994; ). An evolutionary perspective on apoptosis. Cell 76, 777–779.[CrossRef]
    [Google Scholar]
  60. Verma, N. K. & Dey, Ch. S. ( 2004; ). Possible mechanism of miltefosine-mediated death of Leishmania donovani. Antimicrob Agents Chemother 48, 3010–3015.[CrossRef]
    [Google Scholar]
  61. Walker, P. R., Smith, C., Youdale, T., Leblanc, J., Whitfield, J. F. & Sikorska, M. ( 1991; ). Topoisomerase II reactive chemotherapeutic drugs induce apoptosis in thymocytes. Cancer Res 51, 1078–1085.
    [Google Scholar]
  62. Wanderley, J. L., Benjamin, A., Real, F., Bonomo, A., Moreira, M. E. & Barcinski, M. A. ( 2005; ). Apoptotic mimicry: an altruistic behavior in host/Leishmania interplay. Braz J Med Biol Res 38, 807–812.[CrossRef]
    [Google Scholar]
  63. Welburn, S. C., Barcinski, M. A. & Williams, G. T. ( 1997; ). Programmed cell death in trypanosomatids. Parasitol Today 13, 22–26.[CrossRef]
    [Google Scholar]
  64. Williams, G. T. ( 1994; ). Programmed cell death – a fundamental protective response to pathogens. Trends Microbiol 2, 463–464.[CrossRef]
    [Google Scholar]
  65. Wyllie, A. H. ( 1980; ). Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284, 555–556.[CrossRef]
    [Google Scholar]
  66. Wyllie, A. H., Kerr, J. F. R. & Currie, A. R. ( 1980; ). Cell death: the significance of apoptosis. Int Rev Cytol 68, 251–306.
    [Google Scholar]
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