Laser capture microdissection of hepatic stages of the human parasite Plasmodium falciparum for molecular analysis.
Despite the sequencing of parasite genomes and development of DNA microarray technology, gene profiling of parasites remains a difficult task. For example, transcriptome analysis cannot currently be applied to the hepatic stages of the malaria parasite Plasmodium falciparum due to difficulties in obtaining sufficient amounts of parasite material that lies among the large excess of host cell RNA. Here, we describe the isolation of P. falciparum-infected human hepatocytes by a laser capture microdissection approach. Reverse transcriptase polymerase chain reaction amplification of several P. falciparum transcripts demonstrates the high quality of the RNA recovered after microdissection. This approach should enable analysis of P. falciparum transcriptome during its hepatic development, a major step toward the identification of new therapeutic and vaccine targets.
Semblat
Jean-Philippe
JP
Immunobiologie Cellulaire et Moléculaire des Infections Parasitaires, Centre Hospitalier-Universitaire Pitié-Salpêtriere, Université Pierre et Marie Curie, Paris, France.
Silvie
Olivier
O
Franetich
Jean-François
JF
Mazier
Dominique
D
D000818:Animals; D002478:Cells, Cultured; D022781:Hepatocytes / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism / Q000469:parasitology*; D006801:Humans; D007814:Larva / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000254:growth & development* / Q000378:metabolism