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Contribution of Microscopy for Understanding the Mechanism of Action Against Trypanosomatids

  • Esteban Lozano
  • Renata Spina
  • Patricia Barrera
  • Carlos Tonn
  • Miguel A. Sosa
Chapter

Abstract

Transmission electron microscopy (TEM) has proved to be a useful tool to study the ultrastructural alterations and the target organelles of new antitrypanosomatid drugs. Thus, it has been observed that sesquiterpene lactones induce diverse ultrastructural alterations in both T. cruzi and Leishmania spp., such as cytoplasmic vacuolization, appearance of multilamellar structures, condensation of nuclear DNA, and, in some cases, an important accumulation of lipid vacuoles. This accumulation could be related to apoptotic events. Some of the sesquiterpene lactones (e.g., psilostachyin) have also been demonstrated to cause an intense mitochondrial swelling accompanied by a visible kinetoplast deformation as well as the appearance of multivesicular bodies. This mitochondrial swelling could be related to the generation of oxidative stress and associated to alterations in the ergosterol metabolism. The appearance of multilamellar structures and multiple kinetoplasts and flagella induced by the sesquiterpene lactone psilostachyin C indicates that this compound would act at the parasite cell cycle level, in an intermediate stage between kinetoplast segregation and nuclear division. In turn, the diterpene lactone icetexane has proved to induce the external membrane budding on T. cruzi together with an apparent disorganization of the pericellar cytoskeleton. Thus, ultrastructural TEM studies allow elucidating the possible mechanisms and the subsequent identification of molecular targets for the action of natural compounds on trypanosomatids.

Keywords

Natural compounds Terpenes Bioactive molecules Trypanosomatids Trypanosomatid ultrastructure Neglected tropical diseases 

References

  1. Aldunate J, Morello A (1993) Free radicals in the mode of action of parasitic drugs. In: Aruoma OI (ed) Free radicals in tropical diseases. Harwood Academic Publishers, London, pp 137–165Google Scholar
  2. Andrade DV, Gollob KJ, Dutra WO (2014) Acute chagas disease: new global challenges for an old neglected disease. PLoS Negl Trop Dis 8:e3010. https://doi.org/10.1371/journal.pntd.0003010 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Ariyanayagam MR, Fairlamb AH (2001) Ovothiol and trypanothione as antioxidants in trypanosomatids. Mol Biochem Parasitol 115:189–198CrossRefPubMedGoogle Scholar
  4. Ariyanayagam MR, Oza SL, Mehlert A et al (2003) Bis (glutathionyl) spermine and other novel trypanothione analogues in Trypanosoma cruzi. J Biol Chem 278:27612–27619CrossRefPubMedGoogle Scholar
  5. Avila JL, Avila A (1981) Trypanosoma cruzi: allopurinol in the treatment of mice with experimental acute Chagas disease. Exp Parasitol 5:204–208CrossRefGoogle Scholar
  6. Avila JL, Avila A, Monzón H (1984) Differences in allopurinol and 4- aminopyrazolo(3,4-d) pyrimidine metabolism in drug-sensitive and insensitive strains of Trypanosoma cruzi. Mol Biochem Parasitol 1:51–60CrossRefGoogle Scholar
  7. Barrera P, Jimenez-Ortiz V, Giordano O et al (2008) Natural sesquiterpene lactones are active against Leishmania mexicana possible by multiple effects. J Parasitol 94:1143–1149CrossRefPubMedGoogle Scholar
  8. Bergstrom JD, Dufresne C, Bills GF et al (1995) Discovery, synthesis and mechanism of action of the zaragozic acids: potent inhibitors of squalene synthase. Annu Rev Microbiol 49:607–639CrossRefPubMedGoogle Scholar
  9. Bontempi E, Cazzulo J (1990) Digestion of human immunoglobulin G by the major cysteine proteinase (cruzipain) from Trypanosoma cruzi. FEMS Microbiol Lett 58:337–341PubMedGoogle Scholar
  10. Brack C (1968) Electron microscopic studies on the life cycle of Trypanosoma cruzi with special reference to developmental forms in the vector Rhodnius prolixus. Acta Trop 25:289–356PubMedGoogle Scholar
  11. Brengio SD, Belmonte S, Guerreiro E et al (2000) The sesquiterpene lactone dehydroleucodine (DHL) affects the growth of cultured epimastigotes of Trypanosoma cruzi. J Parasitol 86:407–412CrossRefPubMedGoogle Scholar
  12. Campbell DA, Westenberger SJ, Sturm NR (2004) The determinants of Chagas disease: connecting parasite and host genetics. Curr Mol Med 4:549–562CrossRefPubMedGoogle Scholar
  13. Campetella O, Henriksson J, Aslund L et al (1992) The major cysteine proteinase (cruzipain) from Trypanosoma cruzi is encoded by multiple polymorphic tandemly organized genes located on different chromosomes. Mol Biochem Parasitol 50:225–234CrossRefPubMedGoogle Scholar
  14. Costantino VV, Lobos-Gonzalez L, Ibañez J et al (2016) Dehydroleucodine inhibits tumor growth in a preclinical melanoma model by inducing cell cycle arrest, senescence and apoptosis. Cancer Lett 372(1):10–23CrossRefPubMedGoogle Scholar
  15. Da Silva CF, Batista DGJ, De Araújo JS et al (2013) Activities of psilostachyin A and cynaropicrin against Trypanosoma cruzi in vitro and in vivo. Antimicrob Agents Chemother 57(11):5307–5314CrossRefPubMedPubMedCentralGoogle Scholar
  16. De Souza W (2002) Basic cell biology of Trypanosoma cruzi. Curr Pharm Des 8:269–285CrossRefPubMedGoogle Scholar
  17. De Souza W (2008) Electron microscopy of trypanosomes – a historical view. Mem Inst Oswaldo Cruz 103(4):313–325CrossRefPubMedGoogle Scholar
  18. De Souza W (2009) Structural organization of Trypanosoma cruzi. Mem Inst Oswaldo Cruz 104:89–100CrossRefPubMedGoogle Scholar
  19. De Souza W, Cavalcanti DP (2008) DNA-containing organelles in pathogenic protozoa: a review. Trends Cell Mol Biol 2:89–104Google Scholar
  20. De Souza W, Meyer HJ (1975) An electron microscopic and cytochemical study of the cell coat of Trypanosoma cruzi in tissue cultures. Z Parasitenkd 46(3):179–187CrossRefPubMedGoogle Scholar
  21. De Souza W, Martinez-Palomo A, Gonzales-Robbles A (1978) The cell surface of Trypanosoma cruzi: cytochemistry and freeze-fracture. J Cell Sci 33:285–299PubMedGoogle Scholar
  22. Dias JCP (1984) Acute Chagas’ disease. Mem Inst Oswaldo Cruz 79(suppl):85–91CrossRefGoogle Scholar
  23. Dias JCP (1995) Natural history of Chagas’ disease. Arq Bras Cardiol 65:359–366Google Scholar
  24. Docampo R, De Souza W, Miranda K et al (2005) Acidocalcisomes – conserved from bacteria to man. Nat Rev Microbiol 3:251–261CrossRefPubMedGoogle Scholar
  25. Dos Reis F, Judice W, Juliano M et al (2006) The substrate specificity of cruzipain 2, a cysteine protease isoform from Trypanosoma cruzi. FEMS Microbiol Lett 259:215–220CrossRefPubMedGoogle Scholar
  26. Eakin AE, Mills AA, Harth G et al (1992) The sequence, organization, and expression of the major cysteine protease (cruzain) from Trypanosoma cruzi. J Biol Chem 267:7411–7420PubMedGoogle Scholar
  27. El Sayed NM, Myler PJ, Blandin G et al (2005) Comparative genomics of trypanosomatid parasitic Protozoa. Science 309:404–409CrossRefPubMedGoogle Scholar
  28. Elias MCQB, Marques-Porto R, Freymuller E et al (2001) Transcription rate modulation through the Trypanosoma cruzi life cycle occurs in parallel with changes in nuclear organization. Mol Biochem Parasitol 112:79–90CrossRefPubMedGoogle Scholar
  29. Elias MC, Da Cunha JPC, De Faria FP et al (2007) Morphological events during the Trypanosoma cruzi cell cycle. Protist 158:147–157CrossRefPubMedGoogle Scholar
  30. Engel JC, Garcia CT, Hsieh I et al (2000) Upregulation of the secretory pathway in cysteine protease inhibitor-resistant Trypanosoma cruzi. J Cell Sci 113:1345–1354PubMedGoogle Scholar
  31. Garcia ES, Azambuja P (1991) Development and interactions of Trypanosoma cruzi within the insect vector. Parasitol Today 7:240–244CrossRefPubMedGoogle Scholar
  32. Giordano OS, Guerreiro E, Pestchanker MJ et al (1990) The gastric cytoprotective effect of several sesquiterpene lactones. J Nat Prod 53:803–809. https://doi.org/10.1021/np50070a004 CrossRefPubMedGoogle Scholar
  33. Giordano OS, Pestchanker MJ, Guerreiro E et al (1992) Structure-activity relationship in the gastric cytoprotective effect of several sesquiterpene lactones. J Med Chem 35:2452–2458CrossRefPubMedGoogle Scholar
  34. Gonzalez-Pacanowska D, Arison B, Havel CM et al (1988) Isopentenoid synthesis in isolated embryonic Drosophila cells. Farnesol catabolism and v-oxidation. J Biol Chem 263:1301–1306PubMedGoogle Scholar
  35. Gutteridge WE, Davies MJ (1981) Enzymes of purine salvage in Trypanosoma cruzi. FEBS Lett 127:211–214CrossRefPubMedGoogle Scholar
  36. Hammond DJ, Gutteridge WE, Opperdoes FR (1981) A novel location for two enzymes of de novo pyrimidine biosynthesis in trypanosomes and Leishmania. FEBS Lett 128:27–29CrossRefPubMedGoogle Scholar
  37. Jimenez-Ortiz V, Brengio SD, Giordano O et al (2005) The trypanocidal effect of sesquiterpene lactones helenalin and mexicanin on cultured epimastigotes. J Parasitol 91:170–174CrossRefPubMedGoogle Scholar
  38. Johnston DA, Blaxter ML, Degrave WM (1999) Genomics and the biology of parasites. BioEssays 21:131–147CrossRefPubMedGoogle Scholar
  39. de Toledo J, Ambrósio S, Borges C, et al (2014) In Vitro Leishmanicidal Activities of Sesquiterpene Lactones from Tithonia diversifolia against Leishmania braziliensis Promastigotes and Amastigotes. Molecules 19 (5):6070-6079.CrossRefPubMedGoogle Scholar
  40. Kolien AH, Schaub GA (2000) The development of Trypanosoma cruzi in triatomine. Parasitol Today 16:381–387CrossRefGoogle Scholar
  41. Laranja FS, Dias E, Nobrega G et al (1956) Chagas’ disease; a clinical, epidemiologic, and pathologic study. Circulation 14:1035–1060CrossRefPubMedGoogle Scholar
  42. Lazardi K, Urbina JA, De Souza W (1990) Ultrastructural alterations induced by two ergosterol biosynthesis inhibitors, ketoconazole and terbinafine, on epimastigotes and amastigotes of Trypanosoma (Schizotrypanun) cruzi. Antimicrob Agents Chemother 34:2097–2105CrossRefPubMedPubMedCentralGoogle Scholar
  43. Lee KH, Hall IH, Mar EC et al (1977) Sesquiterpene antitumor agents: inhibitors of cellular metabolism. Science 196:533–535CrossRefPubMedGoogle Scholar
  44. Linder JC, Staehelin LA (1977) Plasma membrane specialization in a trypanosomatid flagellate. J Ultrastruct Res 60:246–262CrossRefPubMedGoogle Scholar
  45. Lozano E, Barrera P, Tonn C et al (2012a) The effect of the diterpene 5-epi-icetexone on the cell cycle of Trypanosoma cruzi. Parasitol Int 61:275–279CrossRefPubMedGoogle Scholar
  46. Lozano E, Barrera P, Salinas R et al (2012b) Sesquiterpene lactones and the diterpene 5-epi-icetexone affect the intracellular and extracellular stages of Trypanosoma cruzi. Parasitol Int 61:628–633CrossRefPubMedGoogle Scholar
  47. Lozano ES, Spina RM, Tonn CE et al (2015) An abietane diterpene from Salvia cuspidata and some new derivatives are active against Trypanosoma cruzi. Bioorg Med Chem Lett 25:5481–5484CrossRefPubMedGoogle Scholar
  48. Lozano E, Strauss M, Spina R et al (2016) The in vivo trypanocidal effect of the diterpene 5-epi-icetexone obtained from Salvia gilliesii. Parasitol Int 65:23–26CrossRefPubMedGoogle Scholar
  49. Lukes J, Guilbride DL, Votýpka J et al (2002) Kinetoplast DNA network: evolution of an improbable structure. Eukaryot Cell 1:495–502CrossRefPubMedPubMedCentralGoogle Scholar
  50. Majumder HK (2008) Drug targets in kinetoplastid parasites (Advances in experimental medicine and biology). Vol 625 ISBN: 978-0-387-77569-2 (Print) 978-0-387-77570-8 (Online), Springer Verlag, New YorkGoogle Scholar
  51. Marr JJ, Berens RL, Nelson DJ (1978) Antitrypanosomal effect of allopurinol: conversion in vivo to aminopyrazolopyrimidine nucleotides by Trypanosoma cruzi. Science 201:1018–1020CrossRefPubMedGoogle Scholar
  52. Martinez-Palomo A, De Souza W, Gonzales-Robles AJ (1976) Topographical differences in the distribution of surface coat components and intramembranous particles. J Cell Biol 69:507–513CrossRefPubMedGoogle Scholar
  53. Maya JD, Repetto Y, Agosin M et al (1997) Effects of nifurtimox and benznidazole upon glutathione and trypanothione content in epimastigote, trypomastigote and amastigote forms of Trypanosoma cruzi. Mol Biochem Parasitol 86:101–106PubMedGoogle Scholar
  54. McKerrow J, Caffrey C, Kelly B et al (2006) Proteases in parasitic diseases. Annu Rev Pathol 1:497–536CrossRefPubMedGoogle Scholar
  55. Meirelles M, Juliano L, Carmona E et al (1992) Inhibitors of the major cysteinyl proteinase (GP57/51) impair host cell invasion and arrest the intracellular development of Trypanosoma cruzi in vitro. Mol Biochem Parasitol 52:175–184CrossRefPubMedGoogle Scholar
  56. Meyer H, Porter KR (1954) A study of Trypanosoma cruzi with the electron microscope. Parasitology 44:1–2CrossRefGoogle Scholar
  57. Meyer H, Oliveira Musacchio M, Andrade Mendonça I (1958) Electron microscopy study of Trypanosoma cruzi in thin sections of infected tissue cultures and blood agar forms. Parasitology 48:1–8CrossRefPubMedGoogle Scholar
  58. Miranda K, Benchimol M, Docampo R et al (2000) The fine structure of acidocalcisomes in Trypanosoma cruzi. Parasitol Res 86:373–384CrossRefPubMedGoogle Scholar
  59. Miranda K, Docampo R, Grillo O et al (2004) Acidocalcisomes of trypanosomatids have species-specific elemental composition. Protist 155:395–340CrossRefPubMedGoogle Scholar
  60. Montalvetti A, Rohloff P, Docampo R (2004) A functional aquaporin co-localizes with the vacuolar proton pyrophosphatase to acidocalcisomes and the contractile vacuole complex of Trypanosoma cruzi. J Biol Chem 279:38673–38682CrossRefPubMedGoogle Scholar
  61. Mottram J, Brooks D, Coombs G (1998) Roles of cysteine proteinases of trypanosomes and Leishmania in host-parasite interactions. Curr Opin Microbiol 1:455–460CrossRefPubMedGoogle Scholar
  62. Muschietti LV, Sülsen VP, Martino V (2008) Trypanocidal and leishmanicidal activities of south American medicinal plants. In: Martino VS, Muschietti LV (eds) South American medicinal plants as a potential source of bioactive compounds. Transworld Research, Kerala, pp 149–180Google Scholar
  63. Nieto M, Garcia EE, Giordano OS et al (2000) Icetexane and abietane diterpenoids from Salvia gilliessi. Phytochemistry 53:911–915CrossRefPubMedGoogle Scholar
  64. Ogbadoiyi EO, Robinson DR, Gull K (2003) A high-order trans-membrane structural linkage is responsible for mitochondrial genome positioning and segregation by flagellar basal bodies in trypanosomes. Mol Biol Cell 14:1769–1779CrossRefGoogle Scholar
  65. Opperdoes FR (1987) Compartmentalization of carbohydrate metabolism in trypanosomes. Annu Rev Microbiol 41:127–151CrossRefPubMedGoogle Scholar
  66. Opperdoes FR, Borst P (1977) Localization of nine glycolytic enzymes in a microbody-like organelle in Trypanosoma brucei. FEBS Lett 80:360–364CrossRefPubMedGoogle Scholar
  67. Opperdoes FR, Cotton D (1982) Involvement of the glycosome of Trypanosoma brucei in carbon dioxide fixation. FEBS Lett 143:60–64CrossRefPubMedGoogle Scholar
  68. Penissi AB, Rudolph MI, Piezzi RS (2003) Role of mast cells in gastrointestinal mucosal defense. Biocell 27(2):163–172PubMedGoogle Scholar
  69. Pimenta PF, De Souza WJ (1983) Leishmania mexicana amazonensis: surface charge of amastigote and promastigote forms. Exp Parasitol 56(2):194–206CrossRefPubMedGoogle Scholar
  70. Pinto AY, Ferreira AG Jr, Valente Vda C et al (2009) Urban outbreak of acute Chagas disease in Amazon region of Brazil: four-year follow-up after treatment with benznidazole. Rev Panam Salud Pública 25:77–83CrossRefPubMedGoogle Scholar
  71. Rassi A, Luquetti AO (1992) Therapy of Chagas disease. In: Wendel S, Brener Z, Camargo ME, Rassi A (eds) Chagas disease (American Trypanosomiasis): its impact on transfusion and clinical medicine. ISBT, Sao Paulo, pp 237–247Google Scholar
  72. Sánchez AM, Jiménez-Ortiz V, Sartor T et al (2006) A novel icetexane diterpene, 5-epi-icetexone from Salvia gilliessi is active against Trypanosoma cruzi. Acta Trop 98:118–124CrossRefPubMedGoogle Scholar
  73. Sepúlveda-Boza S, Cassels BK (1996) Plant metabolites active against Trypanosoma cruzi. Planta Med 62:98–105CrossRefPubMedGoogle Scholar
  74. Shapiro TA, Englund PT (1995) The structure and replication of kinetoplast DNA. Annu Rev Microbiol 49:117–143CrossRefPubMedGoogle Scholar
  75. Simpson L (1972) The kinetoplast DNA of the hemoflagellate protozoa. Int Rev Cytol 32:139–207CrossRefGoogle Scholar
  76. Solari AJ (1980) The 3-dimensional fine structure of the mitotic spindle in Trypanosoma cruzi. Chromosoma 78:239–255CrossRefPubMedGoogle Scholar
  77. Souto-Padron T, De Souza W (1978) Ultrastructural localization of basic proteins in Trypanosoma cruzi. J Histochem Cytochem 26:349–356CrossRefPubMedGoogle Scholar
  78. Souto-Padron T, De Souza W (1979) Cytochemical analysis at the fine-structural level of trypanosomatids stained with phosphotungstic acid. J Protozool 26:551–557CrossRefPubMedGoogle Scholar
  79. Souto-Padron T, De Souza W, Heuser JE (1984) Quick-freeze, deep-etch rotary replication of Trypanosoma cruzi and Herpetomonas megaseliae. J Cell Sci 69:167–168PubMedGoogle Scholar
  80. Souto-Padron T, Campetella O, Cazzulo J et al (1990) Cysteine proteinase in Trypanosoma cruzi: immunocytochemical localization and involvement in parasite-host cell interaction. J Cell Sci 96:485–490PubMedGoogle Scholar
  81. De Souza W (1999) A short review on the morphology of Trypanosoma cruzi: from 1909 to 1999. Mem Inst Oswaldo Cruz 94(Suppl) I:17–36CrossRefPubMedGoogle Scholar
  82. Sülsen VP, Güida C, Coussio J et al (2006) In vitro evaluation of trypanocidal activity in plants used in argentine traditional medicine. Parasitol Res 98:370–374CrossRefPubMedGoogle Scholar
  83. Sülsen V, Barrera P, Muschietti L et al (2010) Antiproliferative effect and Ultrastructural alterations induced by Psilostachyin on Trypanosoma cruzi. Molecules 15:545–553. https://doi.org/10.3390/molecules15010545 CrossRefPubMedGoogle Scholar
  84. Sülsen VP, Frank FM, Cazorla SI et al (2011) Psilostachyin C: a natural compound with trypanocidal activity. Int J Antimicrob Agents 37:536–543CrossRefPubMedGoogle Scholar
  85. Tiuman TS, Ueda-Nakamura T, Garcia Cortez DA et al (2005) Antileishmanial activity of parthenolide, a sesquiterpene lactone isolated from Tanacetum parthenium. Antimicrob Agents Chemother 49(1):176–182CrossRefPubMedPubMedCentralGoogle Scholar
  86. Turrens JF (2004) Oxidative stress and antioxidant defenses: a target for the treatment of diseases caused by parasitic protozoa. Mol Asp Med 25:211–220CrossRefGoogle Scholar
  87. Urbina JA (1997) Lipid biosynthesis pathways as chemotherapeutic targets in kinetoplastid parasites. Parasitology 117:S91–S99Google Scholar
  88. Urbina JA (2000) Sterol biosynthesis inhibitors for Chagas’ disease. Curr Opin Anti-Infect Inv Drugs 2:40–46Google Scholar
  89. Urbina JA (2001) Specific treatment of Chagas disease: current status and new developments. Curr Opin Infect Dis 14:733–741CrossRefPubMedGoogle Scholar
  90. Urbina JA (2002) Chemotherapy of Chagas disease. Curr Pharm Des 8:287–295CrossRefPubMedGoogle Scholar
  91. Webster P, Russel DG (1993) The flagellar pocket of trypanosomatids. Parasitol Today 9:201–206CrossRefPubMedGoogle Scholar
  92. Wilkinson SR, Kelly JM (2003) The role of glutathione peroxidases in trypanosomatids. Biol Chem 384:517–525CrossRefPubMedGoogle Scholar
  93. World Health Organization (WHO) (2002) Control of Chagas’ disease. Tech Rep Ser 905:i–vi. World Health Organization, GenevaGoogle Scholar
  94. World Health Organization (WHO) (2017) Chagas disease (American trypanosomiasis). Fact sheetUpdated March 2017. http://www.who.int/mediacentre/factsheets/fs340/en/. Accessed 2 Nov 2017
  95. Zeledón R, Alvarenga NJ, Schosinsky K (1977) Ecology of Trypanosoma cruzi in the insect vector. In: Pan American Health Organization (ed) Chagas’ Disease. Scientific Publication No. 347, Pan American Health Organization, Washington, pp 59–70Google Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Esteban Lozano
    • 1
  • Renata Spina
    • 2
  • Patricia Barrera
    • 2
  • Carlos Tonn
    • 3
  • Miguel A. Sosa
    • 2
  1. 1.Laboratorio de Inmunología y Desarrollo de Vacunas, Instituto de Medicina y Biología Experimental de Cuyo (IMBECU, CCT-CONICET)MendozaArgentina
  2. 2.Laboratorio de Biología y Fisiología Celular Dr. Francisco Bertini, Instituto de Histología y Embriología (IHEM-CONICET)MendozaArgentina
  3. 3.Instituto de Investigación en Tecnología Química (INTEQUI), Facultad de Química Bioquímica y Farmacia, Universidad Nacional de San LuisSan LuisArgentina

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