观察土拉弗朗西斯菌LVS借助脂筏以肌动蛋白为动力被鼠巨噬细胞摄入的过程。细胞胆固醇用菲律平Ⅲ染色,结合神经节苷酯GM1的霍乱毒素B亚基用键合了Alexa 594的兔抗霍乱毒素B亚基二抗显色;肌动蛋白用键合了Alexa 594的鬼笔环肽显色。免疫荧光显微镜观察到脂筏成分中的胆固醇、神经节苷酯GM1均可与细菌共定位;胆固醇可与肌动蛋白共定位。随着感染时间的延长,细菌可离开脂筏。离开脂筏的细菌囊泡可与肌动蛋白共定位。这些发现提示肌动蛋白与脂筏结合,在弗朗西斯菌早期进入巨噬细胞期间发挥重要作用。
The process of the uptake Francisella tularensis LVS by mouse macrophages with the aid of lipid rafts powered by actin was observed.Cell cholesterol was stained with filipin Ⅲ and combined with cholera toxin sub-radical B of gangliosides GM1 were detected and visualized with Alexa Fluor 594 conjugated rabbit-anti-cholera toxin sub-radical B antibodies.Actins were visualized with phalloidin conjugated to Alexa 594.The results indicated that under fluorescence microscope cholesterol,gangliosides GM1 of the component of lipid rafts could be colocalized with the bacterium,and cholesterol could be colocalized with actin.As the prolongation of the infection,bacterium could leave the lipid rafts.Bacterial saccule that left the lipid rafts could be colocalized with actin.These findings suggested that actin combined with lipid rafts played an important role in Francisella early entering into macrophages.