Abstract
We describe a technology for imaging the sequestration of infected red blood cells (iRBC) of the rodent malaria parasite Plasmodium berghei both in the bodies of live mice and in dissected organs, using a transgenic parasite that expresses luciferase. Real-time imaging of sequestered iRBC is performed by measuring bioluminescence produced by the enzymatic reaction in parasites between the luciferase enzyme and its substrate luciferin injected into the mice several minutes prior to imaging. The bioluminescence signal is detected by a sensitive I-CCD photon-counting video camera. Using a reporter parasite that expresses luciferase under the control of a schizont-specific promoter (i.e., the ama-1 promoter), the schizont stage is made visible when detecting bioluminescence signals. Schizont sequestration is imaged during short-term infections with parasites that are synchronized in development or during ongoing infections. Real-time in vivo imaging of iRBC will provide increased insights into the dynamics of sequestration and its role in pathology, and can be used to evaluate strategies that prevent sequestration.
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Acknowledgments
This work was supported by The Netherlands Organization for Scientific Research (ZonMw TOP grant number 9120_6135) and the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 201222.
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Braks, J., Aime, E., Spaccapelo, R., Klop, O., Janse, C.J., Franke-Fayard, B. (2012). Bioluminescence Imaging of P. berghei Schizont Sequestration in Rodents. 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_25
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DOI: https://doi.org/10.1007/978-1-62703-026-7_25
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