原子力显微镜检测α-突触核蛋白转基因鼠线粒体微观结构的变化

Topographic images analysis of mitochondria from α-synuclein transgenic mice utilizing atomic force microscopy

目的使用原子力显微镜检测α-突触核蛋白(α-synuclein,α-syn)转基因小鼠模型中,皮质线粒体表面微观结构的变化。方法提取野生型和转基因小鼠皮质细胞线粒体,使用共聚焦显微镜检测其膜电势变化,使用原子力显微镜的敲击模式检测线粒体膜表面的微观结构。结果 JC-1染色法检测线粒体膜电势结果显示,转基因小鼠线粒体膜电势与野生组相比降低20.4%,差异有统计学意义(P<0.01);原子力显微镜观察结果显示,转基因小鼠线粒体长度与野生组相比增加,表面光滑度减低,线粒体膜孔增多(P<0.05)。结论α-Syn的转基因小鼠线粒体膜电势减低,线粒体膜完整性受到破坏。在原子力显微镜敲击模式下检测离体线粒体,能反映出转基因小鼠线粒体膜表面微观结构的变化。

Objective To investigate the mitochondrial topographic images and nanostructural changes in isolated cortex mitochondria from mice overexpressing human wild-type α-synuclein（ Thy1-α-synuclein mice,Tg/α-syn） via atomic force microscopy（ AFM）.Methods Via JC-1 staining to exam the mitochondrial membrane potential of mitochondria isolated from wild type mice（ WT） and transgenic mice（ TG）. The topographic images and nanostructural changes of mitochondria were detected by AFM between the two groups.Results The mitochondrial membrane potential of mitochondria isolated from transgenic mice was decreased by 20. 4% compared with the wild type mice（ P〈0. 01）. And the topographic images and nanostructure results showed that the length,roughness and number of pore-like structure were increased in mitochondria isolated from TG group（ P〈0. 05）. Conclusion Overexpressing α-syn could attenuate mitochondrial membrane potential（ Δψm） and the mitochondrial membrane was destroyed. The AFM could show the topographic images and nanostructural changes in the isolated mitochondria using tapping mode.