Hostname: page-component-cd9895bd7-q99xh Total loading time: 0 Render date: 2024-12-27T11:15:28.008Z Has data issue: false hasContentIssue false
  • English
  • Français

Complete developmental cycle of Leishmania mexicana in axenic culture

Published online by Cambridge University Press:  06 April 2009

P. A. Bates
P. A. Bates
Affiliation:
Laboratory for Biochemical Parasitology, Department of Zoology, University of Glasgow, Glasgow G12 8QQ
Get access Rights & Permissions [Opens in a new window]

Summary

A complete developmental sequence of Leishmania mexicana has been produced in axenic culture for the first time. This was achieved by manipulation of media, pH and temperature conditions over a period of 16 days. All experiments were initiated with lesion amastigotes that were transformed to multiplicative promastigotes by culture in HOMEM, 10% foetal calf serum, pH 7·5, at 25 °C. Metacyclogenesis was induced by subpassage in Schneider's Drosophila medium, 20% foetal calf serum, pH 5·5, and the resulting forms transformed to axenically growing amastigotes by subpassage in the same medium and raising the temperature to 32 °C. Parasites from each day were characterized with respect to their general morphology using light microscopy of Giemsa-stained smears, and biochemically by analysis of total protein content, proteinases, nucleases and secretory acid phosphatase. The results demonstrated that the three main stages identified - amastigotes, multiplicative promastigotes and metacyclic promastigotes - each exhibited the expected suite of biochemical properties. Further, the changes in morphology observed as the developmental sequence proceeded from stage to stage were accompanied by appropriate changes in biochemical properties. These results provide both useful biochemical markers and a culture system in which to examine the regulation of differentiation and transformation during the Leishmania life-cycle.

Keywords

Leishmaniaaxenic cultureamastigotepromastigotemetacyclicproteinasenucleaseacid phosphatase
Type
Research Article
Information
Parasitology , Volume 108 , Issue 1 , January 1994 , pp. 1 - 9
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Antoine, J. C., Jouanne, C., Ryter, A. & Zilberfarb, V. (1987). Leishmania mexicana: a cytochemical and quantitative study of lysosomal enzymes in infected rat bone marrow-derived macrophages. Experimental Parasitology 64, 485–98.Google Scholar
Bates, P. A. (1993 a). Axenic culture of Leishmania amastigotes. Parasitology Today 9, 143–6.CrossRefGoogle ScholarPubMed
Bates, P. A. (1993 b). Characterization of developmentally-regulated nucleases in promastigotes and amastigotes of Leishmania mexicana. FEMS Microbiology Letters 107, 53–8.CrossRefGoogle ScholarPubMed
Bates, P. A., Hermes, I. & Dwyer, D. M. (1989). Leishmania donovani: immunochemical localization and secretory mechanism of soluble acid phosphatase. Experimental Parasitology 68, 335–46.Google Scholar
Bates, P. A., Robertson, C. D., Tetley, L. & Coombs, G. H. (1992). Axenic cultivation and characterization of Leishmania mexicana amastigote-like forms. Parasitology 105, 193–202.Google Scholar
Bates, P. A. & Tetley, L. (1993). Leishmania mexicana: induction of metacyclogenesis by cultivation of promastigotes at acidic pH. Experimental Parasitology 76, 412–23.Google Scholar
Berens, R. L., Brun, R. & Krassner, S. M. (1976). A simple monophasic medium for axenic culture of hemoflagellates. Journal of Parasitology 62, 360–5.CrossRefGoogle ScholarPubMed
Da Silva, R. & Sacks, D. L. (1987). Metacyclogenesis is a major determinant of Leishmania promastigote virulence and attenuation. Infection and Immunity 55, 2802–6.CrossRefGoogle Scholar
Evans, D. A. (1987). Leishmania. In In Vitro Methods for Parasite Cultivation (ed. Taylor, A. E. R. & Baker, J. R.), pp. 52–75. London: Academic Press.Google Scholar
Gottlieb, M. & Dwyer, D. M. (1982). Identification and partial characterization of an extracellular acid phosphatase activity of Leishmania donovani promastigotes. Molecular and Cellular Biology 2, 76–81.Google Scholar
Hart, D. T., Vickerman, K. & Coombs, G. H. (1981). Transformation in vitro of Leishmania mexicana amastigotes to promastigotes: nutritional requirements and the effect of drugs. Parasitology 83, 529–41.Google Scholar
Ilg, T., Stierhof, Y. -D., Etges, R., Adrian, M., Harbecke, D. & Overath, P. (1991). Secreted acid phosphatase of Leishmania mexicana: a filamentous phosphoglycoprotein polymer. Proceedings of the National Academy of Sciences, USA 88, 8774–8.CrossRefGoogle ScholarPubMed
Killick-Kendrick, R. (1990). The life-cycle of Leishmania in the sandfly with special reference to the form infective to the vertebrate host. Annales de Parasitologic Humaine et Comparée 65 (Suppl. 1), 37–2.Google Scholar
Lawyer, P. G., Young, D. G., Butler, J. F. & Akin, D. E. (1987). Development of Leishmania mexicana in Lutzomyia diabolica and Lutzomyia shannoni (Diptera: Psychodidae). Journal of Medical Entomology 24, 347–55.CrossRefGoogle ScholarPubMed
Lawyer, P. G., Ngumbi, P. M., Anjili, C. O., Odongo, S. O., Mebrahtu, Y. B., Githure, J. I., Koech, D. K. & Roberts, C. R. (1990). Development of Leishmania major in Phlebotomus duboscqi and Sergentomyia schwetzi (Diptera: Psychodidae). American Journal of Tropical Medicine and Hygiene 43, 31–43.CrossRefGoogle ScholarPubMed
Lockwood, B. C., North, M. J., Mallinson, D. J. & Coombs, G. H. (1987). Analysis of Leishmania proteinases reveals developmental changes in species-specific forms and a common 68 kDa activity. FEMS Microbiology Letters 48, 345–50.Google Scholar
Mallinson, D. J. & Coombs, G. H. (1989). Biochemical characteristics of the metacyclic forms of Leishmania major and L. mexicana mexicana. Parasitology 98, 7–15.CrossRefGoogle ScholarPubMed
Maresca, B. & Carratù, L., (1992). The biology of the heat shock response in parasites. Parasitology Today 8, 260–6.CrossRefGoogle ScholarPubMed
Rainey, P. M. & Mackenzie, N. E. (1991). A carbon-13 nuclear magnetic resonance analysis of the products of glucose metabolism in Leishmania pifanoi amastigotes and promastigotes. Molecular and Biochemical Parasitology 45, 307–16.CrossRefGoogle Scholar
Robertson, C. D. & Coombs, G. H. (1990). Characterisation of three groups of cysteine proteinases in the amastigotes of Leishmania mexicana mexicana. Molecular and Biochemical Parasitology 42, 269–76.Google Scholar
Robertson, C. D. & Coombs, G. H. (1992). Stage-specific proteinases of Leishmania mexicana mexicana promastigotes. FEMS Microbiology Letters 94, 127–32.Google Scholar
Russell, D. G., Xu, S. & Chakraborty, P. (1992). Intracellular trafficking and the parasitophorous vacuole of Leishmania mexicana-infected macrophages. Journal of Cell Science 103, 1193–210.Google Scholar
Sacks, D. L. (1989). Metacyclogenesis in Leishmania promastigotes. Experimental Parasitology 69, 100–3.CrossRefGoogle ScholarPubMed
Schneider, P., Rosat, J. -P., Bouvier, J., Louis, J. & Bordier, C. (1992). Leishmania major: differential regulation of the surface metalloprotease in amastigote and promastigote stages. Experimental Parasitology 75, 196–206.CrossRefGoogle ScholarPubMed
Stierhof, Y. -D., Schwarz, H., Menz, B., Russell, D. G., Quinten, M. & Overath, P. (1991). Monoclonal antibodies to Leishmania mexicana promastigote antigens II. Cellular localization of antigens in promastigotes and infected macrophages. Journal of Cell Science 99, 181–6.Google Scholar
Walters, L. L., Modi, G. B., Tesh, R. B. & Burrage, T. (1987). Host-parasite relationship of Leishmania mexicana and Lutzomyia abonnenci (Diptera: Psychodidae). American Journal of Tropical Medicine and Hygiene 36, 294–314.Google Scholar