Department of Epidemiology, University of North Carolina at Chapel Hill, Campus Box 7435, Chapel Hill, NC 27599-7435, USA.
Malar J. 2012 Oct 22;11:350. doi: 10.1186/1475-2875-11-350.
The naturally fluorescent compound quinine has long been used to treat malaria infections. Although some evidence suggests that quinine acts in the parasite food vacuole, the mechanism of action of quinine has not yet been resolved. The Plasmodium falciparum multidrug resistance (pfmdr1) gene encodes a food vacuolar membrane transporter and has been linked with parasite resistance to quinine. The effect of multiple pfmdr1 copies on the subcellular localization of quinine was explored.
Fluorescence microscopy was used to evaluate the subcellular localization of quinine in parasites containing different pfmdr1 copy numbers to determine if copy number of the gene affects drug localization. The acidotropic dye LysoTracker Red was used to label the parasite food vacuole. Time-lapse images were taken to determine quinine localization over time following quinine exposure.
Regardless of pfmdr1 copy number, quinine overlapped with haemozoin but did not colocalize with LysoTracker Red, which labeled the acidic parasite food vacuole.
Quinine localizes to a non-acidic compartment within the food vacuole possibly haemozoin. Pfmdr1 copy number does not affect quinine subcellular localization.
天然荧光化合物奎宁长期以来一直被用于治疗疟疾感染。尽管有一些证据表明奎宁在寄生虫的食物泡中起作用,但奎宁的作用机制尚未解决。恶性疟原虫多药耐药基因 1(pfmdr1)基因编码一种食物泡膜转运蛋白,与寄生虫对奎宁的耐药性有关。本研究探索了多个 pfmdr1 拷贝对奎宁亚细胞定位的影响。
荧光显微镜用于评估含有不同 pfmdr1 拷贝数的寄生虫中奎宁的亚细胞定位,以确定该基因的拷贝数是否影响药物定位。使用酸敏性染料 LysoTracker Red 标记寄生虫的食物泡。拍摄延时图像以确定奎宁暴露后随时间的奎宁定位。
无论 pfmdr1 拷贝数如何,奎宁与疟原虫血红素重叠,但与标记酸性寄生虫食物泡的 LysoTracker Red 不共定位。
奎宁定位于食物泡内的非酸性隔室,可能是疟原虫血红素。pfmdr1 拷贝数不影响奎宁的亚细胞定位。