Mechanisms regulating mitochondrial polarity in neurons: regional distribution and specialization

调节神经元线粒体极性的机制：区域分布和专门化

• 批准号: 9326674
• 负责人: Jason Daniel Vevea
• 金额: $ 5.67万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9326674
• 关键词: Acute; Address; Affect; Age; Axon; Behavior; Biogenesis; Biological Assay; Buffers; Caring; Cell Nucleus; Cell membrane; Cells; Cellular biology; Chemicals; Choline Kinase; Collaborations; Data; Dendrites; Development; Disease; Disease model; Electrophysiology (science); Etiology; Eukaryota; Exhibits; Fibroblasts; Functional disorder; Gene Expression; Glutamates; Glycerophospholipids; Goals; Hippocampus (Brain); Human; In Situ; Individual; Integral Membrane Protein; Knowledge; Length; Literature; Maintenance; Mammalian Cell; Measures; Mediator of activation protein; Membrane Potentials; Membrane Proteins; Mentally Disabled Persons; Messenger RNA; Methods; Microfluidic Microchips; Microfluidics; Mitochondria; Modeling; Molecular; Monitor; Morphology; Motor; Mus; Muscle Fibers; Muscular Dystrophies; Mutation; Nervous system structure; Neurobiology; Neurodegenerative Disorders; Neurons; Nuclear; Optics; Organelles; Output; Oxidation-Reduction; Pathology; Population; Procedures; Proteome; Proteomics; Research; Role; Sampling; Signal Transduction; Site; Sorting - Cell Movement; Stress; Structure; Synaptic Vesicles; Techniques; Technology; Testing; Therapeutic; Toxin; Variant; Vertebral column; Western Blotting; Work; Yeasts; axon injury; base; calcium indicator; cell motility; cell type; clinical care; congenital muscular dystrophy; experimental study; fundamental research; human disease; insight; loss of function; millimeter; mouse model; neuronal cell body; neurotransmission; new technology; next generation; novel; polarized cell; presynaptic; programs; response; retrograde transport; sensor; theories; trafficking; transcriptome sequencing

Project Summary: Neurodegenerative diseases remain some of the most difficult human maladies to understand, much less clinically care for. Fundamental research into basic neurobiology is needed to clarify disease mechanisms and provide research options for possible therapeutic care. Neurons are polarized cells in the nervous system that receive and transmit electrical and chemical information. Many cellular mechanisms support this neuronal function from cytoskeletal structure, nuclear gene expression, to polarized organelle trafficking. Mitochondria are organelles that support a variety of neuronal functions at specialized sites in axons and dendrites and maintain very different dynamics in each compartment. Mitochondrial morphology and trafficking behavior reflect mitochondrial function in a variety cell types and disruption of normal dynamics are related to cellular dysfunction and human disease. Moreover, dysfunctional mitochondria are a hallmark of neurodegenerative diseases. The study of mitochondrial morphology, dynamics, and trafficking in mammalian hippocampal neurons will pave the way forward to a better understanding of how mitochondrial networks are maintained in neurons. This study will focus on determining to what extent mitochondria are specialized in neuronal sub-compartments. Using advanced live cell fluorescent techniques, microfluidic platforms, and next generation proteomics and RNAseq analysis, we will identify the molecular mechanisms behind the formation of mitochondrial polarity and maintenance of mitochondrial networks in mature neurons. Furthermore, literature and our initial insights, support a role for mitofusin 2 in regulating neuronal mitochondrial networks. We are developing knock-OFF technology to study the role of Mfn2 as it pertains to mitochondrial fusion in neurons. Continued development of this new technology will provide a proof of concept and we believe will be generally applicable to the majority of membrane proteins. Transmembrane proteins present a special challenge when trying to acutely inactivate them, there is an urgent need for development of knock-OFF. Furthermore, individual mitochondria from different sub- compartments in neurons will be physically isolated through novel sorting and purification procedures. These mitochondria will be assayed for their proteome to gain insight into how mitochondrial sort and ultimately how the mitochondrial network is maintained. Insights from these studies will be used to understand the neuronal pathology resulting from an atypical congenital muscular dystrophy caused by mutations in choline kinase beta (CHKB).

项目概要： 神经退行性疾病仍然是一些最困难的人类疾病， 更不用说临床护理了。基础神经生物学的基础研究是 需要阐明疾病机制，并为可能的治疗方法提供研究选择。 在乎神经元是神经系统中的极化细胞，其接收和传输电， 化学信息许多细胞机制支持这种神经元功能， 细胞骨架结构，核基因表达，极化细胞器运输。线粒体 是在轴突中的专门位点支持各种神经元功能的细胞器， 树突和维持非常不同的动力学在每个隔室。线粒体形态 和运输行为反映了线粒体在各种细胞类型中的功能， 正常动力学与细胞功能障碍和人类疾病有关。此外，委员会认为， 功能障碍的线粒体是神经变性疾病的标志。研究 哺乳动物海马神经元的线粒体形态、动力学和运输将 为更好地了解线粒体网络如何维持铺平道路 在神经元中。 这项研究将集中在确定线粒体在多大程度上专门用于 神经元亚区室。使用先进的活细胞荧光技术，微流体 平台，下一代蛋白质组学和RNAseq分析，我们将确定分子 线粒体极性形成和维持的机制 成熟神经元中的网络。此外，文献和我们的初步见解，支持的作用， mitofusin 2在调节神经元线粒体网络中的作用。我们正在开发仿制品 技术来研究Mfn 2的作用，因为它涉及神经元中的线粒体融合。继续 这项新技术的发展将提供一个概念证明，我们相信， 一般适用于大多数膜蛋白。跨膜蛋白呈现一种 特别是在试图对他们进行紧急培训时，迫切需要 knock-OFF的发展。此外，来自不同亚细胞的单个线粒体， 神经元中的隔室将通过新的分选和纯化被物理隔离 程序.这些线粒体将被检测其蛋白质组，以深入了解如何 线粒体排序以及最终如何维持线粒体网络。的见解 这些研究将被用来了解非典型的神经病理学， 由胆碱激酶β（CHKB）突变引起的先天性肌营养不良。