title: In vivo imaging of malaria parasites in the murine liver.
abstract: The form of the malaria parasite inoculated by the mosquito, called the sporozoite, transforms inside the host liver into thousands of a new form of the parasite, called the merozoite, which infects erythrocytes. We present here a protocol to visualize in vivo the behavior of Plasmodium berghei parasites in the hepatic tissue of the murine host. The use of GFP-expressing parasites and a high-speed spinning disk confocal microscope allows for the acquisition of four-dimensional images, which provide a time lapse view of parasite displacement and development in tissue volumes. These data can be analyzed to give information on the early events of sporozoite penetration of the hepatic tissue, that is, sporozoite gliding in the liver sinusoids, crossing the sinusoidal barrier, gliding in the parenchyma and traversal of hepatocytes, and invasion of a final hepatocyte, as well as the terminal events of merosome and merozoite release from infected hepatocytes. Combined with the use of mice expressing fluorescent cell types or cell markers, the system will provide useful information not only on the primary infection process, but also on parasite interactions with the host immune cells in the liver.
authors.affiliation: Unité de Biologie et Génétique du Paludisme, Institut Pasteur, 25-28 rue du Dr Roux, 75724 Paris cedex 15, France.
authors.forename: Sabine
authors.initials: S
authors.lastname: Thiberge
authors.forename: Samantha
authors.initials: S
authors.lastname: Blazquez
authors.forename: Patricia
authors.initials: P
authors.lastname: Baldacci
authors.forename: Olivier
authors.initials: O
authors.lastname: Renaud
authors.forename: Spencer
authors.initials: S
authors.lastname: Shorte
authors.forename: Robert
authors.initials: R
authors.lastname: Ménard
authors.forename: Rogerio
authors.initials: R
authors.lastname: Amino
mesh_terms: D000818:Animals; D000852:Anopheles / Q000469:parasitology; D005819:Genetic Markers; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D022781:Hepatocytes / Q000469:parasitology; D008099:Liver / Q000469:parasitology*; D008288:Malaria / Q000469:parasitology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods; D010962:Plasmodium berghei / Q000166:cytology / Q000302:isolation & purification* / Q000472:pathogenicity; D012469:Salivary Glands / Q000166:cytology / Q000469:parasitology
chemical_list: D005819:Genetic Markers; D049452:Green Fluorescent Proteins