Abstract
An increased level of cytosolic free calcium (Ca2+) is an essential second messenger for apical organelle discharge in Plasmodium falciparum merozoites. Here, we describe a method for isolation of viable and invasive P. falciparum merozoites. We also describe methods to measure cytosolic Ca2+ levels in merozoites using fluorescent intracellular calcium indicators such as Fluo-4AM by flow cytometry. Further, we also describe a flow cytometry-based method to detect translocation of apical organelle proteins to the surface of merozoites. Using these methods, we have advanced our understanding of signaling pathways involved in apical organelle discharge during erythrocyte invasion by P. falciparum merozoites.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Aikawa M et al (1978) Erythrocyte entry by malarial parasites. A moving junction between erythrocyte and parasite. J Cell Biol 77:72–82
Dvorak JA et al (1975) Invasion of erythrocytes by malaria merozoites. Science 187:748–750
Miller LH et al (1979) Interaction between cytochalasin-treated malarial parasites and erythrocytes. J Exp Med 49:172–184
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
Cowman AF, Crabb BS (2006) Invasion of red blood cells by malaria parasites. Cell 124:755–766
Singh S et al (2010) Distinct external signals trigger sequential release of apical organelles during erythrocyte invasion by malaria parasites. PLoS Pathog 6:e1000746
Haugland RP (1996) Handbook of fluorescent probes and research biochemicals. Molecular Probes Inc., Eugene, OR, USA
Thomas D et al (2000) A comparison of fluorescent Ca2+ indicator properties and their use in measuring elementary and global Ca2+ signals. Cell Calcium 28:213–223
Di Virgilio F et al (1990) Inhibition of Fura-2 sequestration and secretion with organic anion transport blockers. Cell Calcium 11:57–62
Lupu-Meiri M et al (1993) Calcium entry in Xenopus oocytes: effects of inositol trisphosphate, thapsigargin and DMSO. Cell Calcium 14:101–110
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Singh, S., Chitnis, C.E. (2012). Flow Cytometry-Based Methods for Measurement of Cytosolic Calcium and Surface Protein Expression in Plasmodium falciparum Merozoites. 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_19
Download citation
DOI: https://doi.org/10.1007/978-1-62703-026-7_19
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-025-0
Online ISBN: 978-1-62703-026-7
eBook Packages: Springer Protocols