In vivo investigation of mitochondrial dynamics in the mouse brain

 描述(申请人提供)：神经元是高度分隔的细胞，需要独特的机制将蛋白质和细胞器运输到其特定的隔室。线粒体是调节ATP水平、钙稳态和细胞凋亡的重要细胞器。许多神经退行性疾病，包括肌萎缩侧索硬化症、亨廷顿病、帕金森病和阿尔兹迈尔病，都表现出轴突中线粒体运输的中断，这表明线粒体的定位和功能必须受到高度调控，才能使神经元正常工作。这项研究将开发一种活体方法，通过双光子显微镜对稀疏标记的第2/3层皮质神经元进行活体成像。然后，这种方法将被用于量化线粒体动力学和体内轴突的突触前捕获。神经元活动的作用也将被询问，以确定它对线粒体动力学、突触前捕获和功能的影响。最后，这项研究将开发一种方法，与体树突状线粒体相比，无偏见地识别轴突线粒体中独特或丰富的蛋白质。这项研究的结果将为了解线粒体动力学的体内机制以及活动如何调节它们的突触前捕获和功能提供新的见解。

 DESCRIPTION (provided by applicant): Neurons are highly compartmentalized cells that require unique mechanisms for trafficking proteins and organelles to their specific compartments. Mitochondria are vital organelles that regulate the levels of ATP, calcium homeostasis and apoptosis. Many neurodegenerative diseases including ALS, Huntington's, Parkinson's and Alzhemier's show a disruption of mitochondrial transport in the axon suggesting that mitochondrial localization and function must be highly regulated for the neuron to function properly. This study will develop an in vivo method for the live imaging of sparsely labeled layer 2/3 cortical neurons via 2-photon microscopy. This method will then be used to quantitate mitochondrial dynamics and presynaptic capture along axons in vivo. The role of neuronal activity will also be interrogated to determine the effect it has on mitochondrial dynamics, presynaptic capture and function. Finally, this study will develop a method for the unbiased identification of proteins unique or abundant in axonal mitochondria as compared to somato-dendritic mitochondria. The results of this study will provide new insights into the in vivo mechanisms of mitochondrial dynamics and how activity regulates their presynaptic capture and function.