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Static and Dynamic Imaging of Erythrocyte Invasion and Early Intra-erythrocytic Development in Plasmodium falciparum

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Malaria

Part of the book series: Methods in Molecular Biology ((MIMB,volume 923))

Abstract

Cellular imaging has reemerged in recent years as a powerful approach to provide researchers with a direct measure of essential molecular events in a cell’s life, ranging in scale from broad morphological observations of whole cells to intricate single molecule imaging. When combined with quantitative image analysis, the available imaging techniques can act as a critical means to confirm hypotheses, drive the formation of new theories or provide accurate determination of protein localization at subcellular and nanometer scales. Here, we describe two methodological approaches for imaging the transient step of malaria parasite invasion of the human erythrocyte. When applied to image the most virulent human malaria parasite, Plasmodium falciparum, the first approach, using live time-lapse wide-field microscopy, allows the capture of transient events during invasion and postinvasion intra-erythrocytic development, while the second, using immunofluorescence assay (IFA) of fixed samples, allows high-definition exploration of parasite architecture on multiple platforms.

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References

  1. Trager W (1956) The intracellular position of malarial parasites. Trans R Soc Trop Med Hyg 50:419–420

    Article  PubMed  CAS  Google Scholar 

  2. Huff CG et al (1960) The morphology and behavior of living exoerythrocytic stages of Plasmodium gallinaceum and P. fallax and their host cells. J Biophys Biochem Cytol 7:93–102

    Article  PubMed  CAS  Google Scholar 

  3. Ladda R et al (1969) Penetration of erythrocytes by merozoites of mammalian and avian malarial parasites. J Parasitol 55:633–644

    Article  PubMed  CAS  Google Scholar 

  4. Dvorak JA et al (1975) Invasion of erythrocytes by malaria merozoites. Science 187:748–750

    Article  PubMed  CAS  Google Scholar 

  5. Gilson PR, Crabb BS (2009) Morphology and kinetics of the three distinct phases of red blood cell invasion by Plasmodium falciparum merozoites. Int J Parasitol 39:91–96

    Article  PubMed  CAS  Google Scholar 

  6. Glushakova S et al (2009) Irreversible effect of cysteine protease inhibitors on the release of malaria parasites from infected erythrocytes. Cell Microbiol 11:95–105

    Article  PubMed  CAS  Google Scholar 

  7. Glushakova S et al (2005) Membrane transformation during malaria parasite release from human red blood cells. Curr Biol 15:1645–1650

    Article  PubMed  CAS  Google Scholar 

  8. Richard D et al (2009) Identification of rhoptry trafficking determinants and evidence for a novel sorting mechanism in the malaria parasite. PLoS Pathog 5:e1000328

    Article  PubMed  Google Scholar 

  9. Treeck M et al (2009) Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process. PLoS Pathog 5:e1000322

    Article  PubMed  Google Scholar 

  10. Boyle MJ et al (2010) Interactions with heparin-like molecules during erythrocyte invasion by Plasmodium falciparum merozoites. Blood 115:4559–4568

    Article  PubMed  CAS  Google Scholar 

  11. Langreth SG et al (1978) Plasmodium falciparum: merozoite invasion in vitro in the ­presence of chloroquine. Exp Parasitol 46:235–238

    Article  PubMed  CAS  Google Scholar 

  12. Bannister LH et al (1975) Structure and invasive behaviour of Plasmodium knowlesi merozoites in vitro. Parasitology 71:483–491

    Article  PubMed  CAS  Google Scholar 

  13. Aikawa M et al (1978) Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite. J Cell Biol 77:72–82

    Article  PubMed  CAS  Google Scholar 

  14. Boyle MJ et al (2010) Isolation of viable Plasmodium falciparum merozoites to define erythrocyte invasion events and advance vaccine and drug development. Proc Natl Acad Sci USA 107:14378–14383

    Article  PubMed  CAS  Google Scholar 

  15. Riglar DT et al (2011) Super-resolution dissection of coordinated events during malaria parasite invasion of the human erythrocyte. Cell Host Microbe 9:9–20

    Article  PubMed  CAS  Google Scholar 

  16. Schermelleh L et al (2010) A guide to super-resolution fluorescence microscopy. J Cell Biol 190:165–175

    Article  PubMed  CAS  Google Scholar 

  17. Hanssen E et al (2010) Whole cell imaging reveals novel modular features of the exomembrane system of the malaria parasite, Plasmodium falciparum. Int J Parasitol 40:123–134

    Article  PubMed  CAS  Google Scholar 

  18. Yeoman JA et al (2011) Tracking glideosome-associated protein 50 reveals the development and organization of the inner membrane complex of Plasmodium falciparum. Eukaryot Cell 10:556–564

    Article  PubMed  CAS  Google Scholar 

  19. Bannister LH et al (2000) A brief illustrated guide to the ultrastructure of Plasmodium falciparum asexual blood stages. Parasitol Today 16:427–433

    Article  PubMed  CAS  Google Scholar 

  20. Abkarian M et al (2011) A novel mechanism for egress of malarial parasites from red blood cells. Blood 117:4118–4124

    Article  PubMed  CAS  Google Scholar 

  21. Pachlatko E et al (2010) MAHRP2, an exported protein of Plasmodium falciparum, is an essential component of Maurer’s cleft tethers. Mol Microbiol 77:1136–1152

    Article  PubMed  CAS  Google Scholar 

  22. Richard D et al (2010) Interaction between Plasmodium falciparum apical membrane antigen 1 and the rhoptry neck protein complex defines a key step in the erythrocyte invasion process of malaria parasites. J Biol Chem 285:14815–14822

    Article  PubMed  CAS  Google Scholar 

  23. Grüring C et al (2011) Development and host cell modifications of Plasmodium falciparum blood stages in four dimensions. Nat Commun 2:165

    Article  PubMed  Google Scholar 

  24. Tilley L et al (2007) Illuminating Plasmodium falciparum-infected red blood cells. Trends Parasitol 23:268–277

    Article  PubMed  Google Scholar 

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Acknowledgments

We apologize to many researchers in this field whose work we have not been able to cite directly because of the limits of space. We thank James Beeson and Michelle Boyle for their helpful comments. This work was supported by the National Health and Medical Research Council of Australia (637340). DTR is supported through a Pratt Foundation postgraduate scholarship from the University of Melbourne. JB is supported by a Future Fellowship (FT100100112) from the Australian Research Council.

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Correspondence to Jake Baum .

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Riglar, D.T., Baum, J. (2012). Static and Dynamic Imaging of Erythrocyte Invasion and Early Intra-erythrocytic Development in Plasmodium falciparum . In: Ménard, R. (eds) Malaria. Methods in Molecular Biology, vol 923. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-026-7_18

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  • DOI: https://doi.org/10.1007/978-1-62703-026-7_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-025-0

  • Online ISBN: 978-1-62703-026-7

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