Skip to main content

Employing Transgenic Parasite Strains to Study the Ca2+ Dynamics in the Human Malaria Parasite Plasmodium falciparum

  • Protocol
  • First Online:
Calcium Signalling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1925))

Abstract

Studying Ca2+ dynamics in protozoan parasites is not an easy task. Loading of parasites with commonly used Ca2+ fluorescent dyes (such as Fuo4-AM) remains as the major protocol to measure the Ca2+ oscillations inside the cell. In this chapter, we describe an alternative method to study Ca2+ signaling in Plasmodium falciparum parasite. This method employs the construction of transgenic parasites (through standard molecular biology techniques), selection of the transfected population, and use of those parasites in spectrofluorometric Ca2+ assays.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

eBook
USD 44.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 59.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Berridge MJ, Lipp P et al (2000) The versatility and universality of calcium signalling. Nat Rev Mol Cell Biol 1:11–21

    Article  CAS  Google Scholar 

  2. Plattner H, Sehring IM et al (2012) Calcium signaling in closely related protozoan groups (Alveolata): non-parasitic ciliates (Paramecium, Tetrahymena) vs. parasitic Apicomplexa (Plasmodium, Toxoplasma). Cell Calcium 51:351–382

    Article  CAS  Google Scholar 

  3. Moreno SN, Docampo R (2003) Calcium regulation in protozoan parasites. Curr Opin Microbiol 6:359–364

    Article  CAS  Google Scholar 

  4. Budu A, Garcia CR (2012) Generation of second messengers in Plasmodium. Microbes Infect 14:787–795

    Article  CAS  Google Scholar 

  5. Grynkiewicz G, Poenie M et al (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    CAS  PubMed  Google Scholar 

  6. Borges-Pereira L, Budu A et al (2015) Calcium signaling throughout the toxoplasma gondii lytic cycle: a study using genetically encoded calcium indicators. J Biol Chem 290:26914–26926

    Article  CAS  Google Scholar 

  7. Borges-Pereira L, Campos BR et al (2014) The GCaMP3—a GFP-based calcium sensor for imaging calcium dynamics in the human malaria parasite Plasmodium falciparum. Methods 1:151–154

    Google Scholar 

  8. Tian L, Hires SA et al (2012) Imaging neuronal activity with genetically encoded calcium indicators. Cold Spring Harb Protoc 2012:647–656

    Article  Google Scholar 

  9. Mank M, Griesbeck O (2008) Genetically encoded calcium indicators. Chem Rev 108:1550–1564

    Article  CAS  Google Scholar 

  10. Fidock DA, Nomura T et al (2000) Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance. Mol Cell 6:861–871

    Article  CAS  Google Scholar 

  11. Trager W, Jensen JB (1976) Human malaria parasites in continuous culture. Science 193:673–675

    Article  CAS  Google Scholar 

  12. Lambros C, Vanderberg JP (1979) Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol 65:418–420

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), process 2011/51295-5 to C.R.S.G and 2016/14411-0 to L.B.P.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Célia R. S. Garcia .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Borges-Pereira, L., Garcia, C.R.S. (2019). Employing Transgenic Parasite Strains to Study the Ca2+ Dynamics in the Human Malaria Parasite Plasmodium falciparum . In: Raffaello, A., Vecellio Reane, D. (eds) Calcium Signalling. Methods in Molecular Biology, vol 1925. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9018-4_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9018-4_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9017-7

  • Online ISBN: 978-1-4939-9018-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics