Abstract
Plasmodium sporozoites are the infectious, highly motile forms of the malaria parasite transmitted by Anopheles mosquitoes. Sporozoite motility can be assessed following the dissection of Anopheles salivary glands and isolation of sporozoites in vitro.
Keywords: Plasmodium, Plasmodium berghei, Sporozoites, Salivary gland isolation, Dissection, Gliding motility, Malaria, Mosquitoes, Anopheles
Background
Sporozoites of the phylum Plasmodium, the causative agent of malaria, are transmitted into the skin of their vertebrate host through the bite of an infectious mosquito. Sporozoite motility is a key prerequisite for parasite transmission and successful infection of the vertebrate host. Motility constitutes the first parasite mechanism that can be inhibited and is thus of interest for intervention strategies. Genetic modifications affecting gliding motility or motility modulating compounds can be readily investigated using 2D in vitro assays.
Part I: Isolation of salivary gland sporozoites
Materials and Reagents
Equipment
Procedure
Part II: Motility assay
Software
1. Fiji (download at http://imagej.net/Fiji)
Note: If using sporozoites from 5 or less well-infected female mosquitoes gliding assays can be performed directly. Accudenz purification of sporozoites (Kennedy et al., 2012) is needed when sporozoites derived from more than 5 salivary glands are used due to impurity with mosquito debris.
Data analysis
Notes
Recipes
Acknowledgments
Funding: collaborative research center SFB 1129 of the Deutsche Forschungsgemeinschaft.
References
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