In vivo investigation of mitochondrial dynamics in the mouse brain

小鼠大脑线粒体动力学的体内研究

• 批准号: 8869236
• 负责人: Tommy L Lewis
• 金额: $ 9.5万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-15 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8869236
• 关键词: Affect; Alzheimer' s Disease; Apoptosis; Axon; Biosensor; Brain; Calcium; Cells; Cephalic; Complex; Data; Dendrites; Development; Electroporation; Fluorescence-Activated Cell Sorting; Fluorescent Probes; Glucose; Head; Homeostasis; Huntington Disease; Image; Investigation; Knockout Mice; Label; Laser Scanning Microscopy; Life; Maintenance; Mass Spectrum Analysis; Measurement; Measures; Membrane Potentials; Messenger RNA; Metabolism; Methods; Microscopy; Mitochondria; Mitochondrial Proteins; Monitor; Mus; Neurodegenerative Disorders; Neurons; Organelles; Parkinson Disease; Pathway interactions; Phosphotransferases; Play; Presynaptic Terminals; Production; Proteins; Proteomics; Resolution; Rhodopsin; Role; STK11 gene; Sorting - Cell Movement; Staging; Synapses; Testing; hippocampal pyramidal neuron; imaging modality; in utero; in vivo; in vivo imaging; insight; mitochondrial membrane; neurotransmitter release; novel; novel strategies; optogenetics; postnatal; presynaptic; protein transport; public health relevance; research study; tool; trafficking; two-photon

 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.

 描述（由申请人提供）：神经元是高度区室化的细胞，需要独特的机制将蛋白质和细胞器运输到其特定的区室。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 ALS、亨廷顿舞蹈症、帕金森舞蹈症和阿尔茨海默病，显示轴突中线粒体运输的破坏，这表明线粒体定位和功能必须受到高度调节，神经元才能正常发挥作用。本研究将开发一种通过 2 光子显微镜对稀疏标记的 2/3 层皮质神经元进行实时成像的体内方法。然后，该方法将用于定量体内沿轴突的线粒体动力学和突触前捕获。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，本研究将开发一种方法，用于公正地鉴定轴突线粒体中与体细胞树突状线粒体相比独特或丰富的蛋白质。这项研究的结果将为线粒体动力学的体内机制以及活性如何调节其突触前捕获和功能提供新的见解。