Advertisement

Synchronization of Pathogenic Protozoans

  • Staffan SvärdEmail author
  • Karin Troell
Part of the Methods in Molecular Biology book series (MIMB, volume 761)

Abstract

Protozoans are single-cell eukaryotes and many of the best studied protozoans are parasitic to humans (e.g., Plasmodium falciparum causing malaria and Trypanosoma brucei causing sleeping sickness). These organisms are distantly related to humans but with retained eukaryotic type of cellular processes, making them good model systems for studies of the evolution of basic processes like the cell cycle. Giardia intestinalis causes 250 million cases of diarrhea yearly and is one of the earliest diverging protozoans. It has recently been possible to synchronize its cell cycle using compounds that inhibit different steps of the cell cycle and the detailed protocol is described here.

Key words

Cell cycle malaria parasite aphidicolin Giardia 

References

  1. 1.
    Morrison, H. G., McArthur, A. G., Gillin, F. D., Aley, S. B., Adam, R. D., Olsen, G. J., et al. (2007) Genomic minimalism in the early diverging intestinal parasite Giardia lamblia. Science 317, 1921–1926.PubMedCrossRefGoogle Scholar
  2. 2.
    Gardner, M. J., Hall, N., Fung, E., White, O., Berriman, M., Hyman, R. W., et al. (2002) Genome sequence of the human malaria parasite Plasmodium falciparum. Nature 419, 498–511.PubMedCrossRefGoogle Scholar
  3. 3.
    Reilly, H. B., Wang, H., Steuter, J. A., Marx, A. M., and Ferdig, M. T. (2007) Quantitative dissection of clone-specific growth rates in cultured malaria parasites. Int. J. Parasitol. 37, 1599–1607.PubMedCrossRefGoogle Scholar
  4. 4.
    Inselburg, J., and Banyal, H. S. (1984) Synthesis of DNA during the asexual cycle of Plasmodium falciparum in culture. Mol. Biochem. Parasitol. 10, 79–87.PubMedCrossRefGoogle Scholar
  5. 5.
    Kutner, S., Breuer, W. V., Ginsburg, H., Aley, S. B., and Cabantchik, Z. I. (1985) Characterization of permeation pathways in the plasma membrane of human erythrocytes infected with early stages of Plasmodium falciparum: association with parasite development. J. Cell Physiol. 125, 521–527.PubMedCrossRefGoogle Scholar
  6. 6.
    Rojas, M. O., and Wasserman, M. (1993) Effect of low temperature on the in vitro growth of Plasmodium falciparum. J. Eukaryot. Microbiol. 40, 149–152.PubMedCrossRefGoogle Scholar
  7. 7.
    Lambros, C., and Vanderberg, J. P. (1979) Synchronization of Plasmodium falciparum erythrocytic stages in culture. J. Parasitol. 65, 418–420.PubMedCrossRefGoogle Scholar
  8. 8.
    Bhakdi, S. C., Ottinger, A., Somsri, S., Sratongno, P., Pannadaporn, P., Chimma, P., et al. (2010) Optimized high gradient magnetic separation for isolation of Plasmodium-infected red blood cells. Malar. J. 9, 38.PubMedCrossRefGoogle Scholar
  9. 9.
    Naughton, J. A., and Bell, A. (2007) Studies on cell-cycle synchronization in the asexual erythrocytic stages of Plasmodium falciparum. Parasitology 134, 331–337.PubMedCrossRefGoogle Scholar
  10. 10.
    Radfar, A., Mendez, D., Moneriz, C., Linares, M., Marín-García, P., Puyet, A., et al. (2009) Synchronous culture of Plasmodium falciparum at high parasitemia levels. Nat. Protoc. 4, 1828–1844.CrossRefGoogle Scholar
  11. 11.
    Mead, P. S., Slutsker, L., Griffin, P. M., and Tauxe, R. V. (1999) Food-related illness and death in the United States reply to Dr. Hedberg. Emerg. Infect. Dis. 5, 841–842.PubMedCrossRefGoogle Scholar
  12. 12.
    Taylor, S., Barragan, A., Su, C., Fux, B., Fentress, S. J., Tang, K., et al. (2006) A secreted serine-threonine kinase determines virulence in the eukaryotic pathogen Toxoplasma gondii. Science 314, 1776–1780.PubMedCrossRefGoogle Scholar
  13. 13.
    Radke, J. R., and White, M. W. (1998) A cell cycle model for the tachyzoite of Toxoplasma gondii using the Herpes simplex virus thymidine kinase. Mol. Biochem. Parasitol. 94, 237–247.PubMedCrossRefGoogle Scholar
  14. 14.
    Conde de Felipe, M. M., Lehmann, M. M., Jerome, M. E., and White, M. W. (2008) Inhibition of Toxoplasma gondii growth by pyrrolidine dithiocarbamate is cell cycle specific and leads to population synchronization. Mol. Biochem. Parasitol. 157, 22–31.PubMedCrossRefGoogle Scholar
  15. 15.
    Simpson, L., and Braly, P. (1970) Synchronization of Leishmania tarentolae by hydroxyurea. J. Protozool. 17, 511–517.PubMedGoogle Scholar
  16. 16.
    Galanti, N., Dvorak, J. A., Grenet, J., and McDaniel, J. P. (1994) Hydroxyurea-induced synchrony of DNA replication in the Kinetoplastida. Exp. Cell Res. 214, 225–230.PubMedCrossRefGoogle Scholar
  17. 17.
    Gale, M., Jr., Carter, V., and Parsons, M. (1994) Cell cycle-specific induction of an 89 kDa serine/threonine protein kinase activity in Trypanosoma brucei. J. Cell Sci. 107, 1825–1832.PubMedGoogle Scholar
  18. 18.
    Jantzen, H., Schulze, I., and Stohr, M. (1988) Relationship between the timing of DNA replication and the developmental competence in Acanthamoeba castellanii. J. Cell Sci. 91, 389–399.PubMedGoogle Scholar
  19. 19.
    Svard, S. G., Hagblom, P., and Palm, J. E. (2003) Giardia lamblia – a model organism for eukaryotic cell differentiation. FEMS Microbiol. Lett. 218, 3–7.PubMedCrossRefGoogle Scholar
  20. 20.
    Poxleitner, M. K., Carpenter, M. L., Mancuso, J. J., Wang, C. J., Dawson, S. C., and Cande, W. Z. (2008) Evidence for karyogamy and exchange of genetic material in the binucleate intestinal parasite Giardia intestinalis. Science 319, 1530–1533.PubMedCrossRefGoogle Scholar
  21. 21.
    Roxstrom-Lindquist, K., Palm, D., Reiner, D., Ringqvist, E., and Svard, S. G. (2006) Giardia immunity – an update. Trends Parasitol. 22, 26–31.PubMedCrossRefGoogle Scholar
  22. 22.
    Reiner, D. S., Ankarklev, J., Troell, K., Palm, D., Bernander, R., Gillin, F. D., et al. (2008) Synchronisation of Giardia lamblia: identification of cell cycle stage-specific genes and a differentiation restriction point. Int. J. Parasitol. 38, 935–944.PubMedCrossRefGoogle Scholar
  23. 23.
    Poxleitner, M. K., Dawson, S. C., and Cande, W. Z. (2008) Cell cycle synchrony in Giardia intestinalis cultures achieved by using nocodazole and aphidicolin. Eukaryot. Cell 7, 569–574.PubMedCrossRefGoogle Scholar
  24. 24.
    Cooper, S., Iyer, G., Tarquini, M., and Bissett, P. (2006) Nocodazole does not synchronize cells: implications for cell-cycle control and whole-culture synchronization. Cell Tissue Res. 324, 237–242.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Cell and Molecular BiologyUppsala UniversityUppsalaSweden
  2. 2.National Veterinary InstituteUppsalaSweden

Personalised recommendations