Plasmodium berghei: quantitation of in vitro effects of antimalarial drugs on exoerythrocytic development by a ribosomal RNA probe.
A stage-specific ribosomal RNA probe has been used to quantitate exoerythrocytic development of Plasmodium berghei in primary cultures of mouse hepatocytes. Parasite rRNA could be detected as soon as 6 hr after sporozoite invasion and was increased during schizogony to a maximum at 48 hr, when mature schizonts were identified by microscopy. As few as 10 exoerythrocytic schizonts could be detected by filter blot hybridization, followed by autoradiography and liquid scintillation counting. By hybridizing the culture rRNA samples with either parasite-specific or universal rRNA probes, the in vitro tissue schizonticidal activity and hepatotoxicity of primaquine, two of its analogues, and pyrimethamine, could be assessed. After a 48-hr exposure of the culture to serial dilutions of each drug, a quantitative relationship was demonstrated between the decrease of the parasite rRNA and the increase of the drug concentrations. No significant parasite-specific rRNA could be detected at the concentration achieving complete inhibition of schizont formation but causing no cytotoxic effects on host hepatocytes. In contrast to microscopic-based assays, this molecular approach provides an objective and quantitative in vitro method for rapid screening and evaluation of tissue schizonticidal antimalarials.
Li
J
J
Second Military Medical University, Shanghai, People's Republic of China.
Zhu
J D
JD
Appiah
A
A
McCutchan
T F
TF
Long
G W
GW
Milhous
W K
WK
Hollingdale
M R
MR
D000634:Aminoquinolines / Q000494:pharmacology; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D001483:Base Sequence; D002478:Cells, Cultured; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000469:parasitology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010962:Plasmodium berghei / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D011319:Primaquine / Q000031:analogs & derivatives / Q000494:pharmacology; D011739:Pyrimethamine / Q000494:pharmacology; D015347:RNA Probes; D016053:RNA, Protozoan / Q000032:analysis*; D012335:RNA, Ribosomal / Q000032:analysis*
D000634:Aminoquinolines; D000962:Antimalarials; D015347:RNA Probes; D016053:RNA, Protozoan; D012335:RNA, Ribosomal; C055852:tafenoquine; C068820:WR 242511; D011319:Primaquine; D011739:Pyrimethamine