Monitoring the Uptake of Glycosphingolipids In Plasmodium falciparum-Infected Erythrocytes Using Both Fluorescence Microscopy and Capillary Electrophoresis with Laser-Induced Fluorescence Detection

摘要

The metabolism of glycosphingolipids by the malariacausingnparasite Plasmodium falciparum plays an importantnrole in the progression of the disease. We report annew and highly sensitive method to monitor the uptakenof glycosphingolipids in infected red blood cells (iRBCs).nA tetramethylrhodamine-labeled glycosphingolipid (GM1-nTMR) was used as a substrate. Uptake was demonstratednby fluorescence microscopy. The iRBCs were lysed withna 15% solution of saponin and washed with phosphatenbuffered saline to release intact parasites. The parasitesnwere further lysed and the resulting homogenates werenanalyzed by capillary electrophoresis with laser-inducednfluorescence detection. The lysate from erythrocytes infectednat 1% parasitemia generated a signal 20 standardndeviations larger than uninfected erythrocytes, whichnsuggests that relatively low infection levels can be studiednwith this technique.