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).

项目摘要： 神经退行性疾病仍然是最困难的人类疾病 了解，更不用说临床照顾了。基本神经生物学的基本研究是 需要澄清疾病机制并为可能的治疗提供研究选择 关心。神经元是神经系统中的极化细胞，接收和传输电气， 化学信息。许多细胞机制支持此神经元功能 细胞骨架结构，核基因表达，用于两极分化的细胞器运输。线粒体 是在轴突和 树突并保持每个隔间中的动态非常不同。线粒体形态 贩运行为反映了各种细胞类型中的线粒体功能和破坏 正常动力学与细胞功能障碍和人类疾病有关。而且， 线粒体功能失调是神经退行性疾病的标志。研究 哺乳动物海马神经元中的线粒体形态，动力学和贩运 铺平道路，以更好地了解如何维护线粒体网络 在神经元中。 这项研究将着重于确定线粒体在多大程度上专门从事 神经元子室。使用高级活细胞荧光技术，微流体 平台以及下一代蛋白质组学和RNASEQ分析，我们将确定分子 线粒体极性形成的机制和线粒体的维持 成熟神经元中的网络。此外，文学和我们的最初见解支持 丝线2在调节神经元线粒体网络中。我们正在发展仿制 研究MFN2与神经元中线粒体融合有关的技术。持续 这项新技术的开发将提供概念验证，我们相信 通常适用于大多数膜蛋白。跨膜蛋白呈现 试图急性灭活它们时，特别需要挑战 仿冒品的发展。此外，来自不同子的单个线粒体 神经元中的隔室将通过新颖的分类和纯化进行物理隔离 程序。这些线粒体将被分析，以了解其蛋白质组，以深入了解如何 线粒体排序，最终如何保持线粒体网络。来自 这些研究将用于了解非典型的神经元病理 由胆碱激酶β（CHKB）突变引起的先天性肌肉营养不良。