Regulation of cargo transport during neuronal development and disease

神经元发育和疾病期间货物运输的调节

• 批准号: 10863335
• 负责人: MARY C HALLORAN
• 金额: $ 53.57万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10863335
• 关键词: Adaptor Signaling Protein; Afferent Neurons; Axon; Behavior; Binding; Biosensor; C-terminal; Calcium; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Co-Immunoprecipitations; Complex; Cues; Data; Defect; Development; Developmental Process; Disease; Embryo; Endosomes; Environment; Genes; Goals; Health; Human; Image; Individual; Kinesin; Knowledge; Light; Location; Lysosomes; Maintenance; Mediating; Missense Mutation; Mitochondria; Modeling; Molecular; Molecular Target; Morphogenesis; Morphology; Motor; Muscle fasciculation; Mutagenesis; Mutation; Natural regeneration; Neurodegenerative Disorders; Neurons; Organelles; Oxidation-Reduction; Peripheral; Phenotype; Promoter Regions; Proteins; Regulation; Role; Sensory; Shapes; Signal Transduction; Site; Spastic Paraplegia; Specificity; Structure; Symptoms; Syndrome; Tertiary Protein Structure; Testing; Zebrafish; axon growth; axon injury; cellular pathology; developmental disease; early onset; experimental study; extracellular; human disease; human model; in vivo; in vivo Model; in vivo imaging; infancy; insight; link protein; mutant; neural; neurodevelopment; neuron development; novel strategies; overexpression; sensory neuropathy; somatosensory

Project summary Development of highly elaborate and polarized neuronal morphology requires formation of molecularly distinct compartments within neurons and precise subcellular localization of proteins and organelles. Neuronal compartmentalization requires that motors such as kinesin-1 deliver specific cargos to different sites within the neuron. Neurons are especially dependent on diverse and high fidelity cargo transport because of their highly polarized and complex structure. Defects in transport underlie multiple human developmental and neurodegenerative diseases. Kinesin-1 is known to mediate long-distance transport of multiple cargos, but the mechanisms that determine specificity of cargo selection and delivery during development are poorly understood. The cargo-binding kinesin light chain (KLC) subunits of kinesin-1 (encoded by klc1-4 genes) are likely involved in specificity, yet the developmental processes they mediate are not understood. This is a critical knowledge gap, as mutations in human klc2 and klc4 genes cause spastic paraplegia diseases with very early onset. A major challenge to the field and the long term goal of this project is to understand the mechanisms controlling cargo transport and localization as neurons develop in their natural environment, where they must integrate multiple extracellular cues. We established a model in which we can image dynamics of neuronal cargo transport within developing sensory neurons in the intact zebrafish embryo. Vertebrate sensory neurons extend distinct central and peripheral axons to form the sensory circuit. We discovered unique functions for individual KLCs during neural development, including roles in shaping axon arbors, axon branching, guidance and maintenance during development, and roles in circuit function. In Aim 1 we will determine the mechanisms by which KLC4 regulates axon branching and compartmentalization, and the organelle cargos it transports. In Aim 2 we will determine functions of KLC2 in axon development and cargo transport, and mechanisms underlying human SPOAN syndrome. In Aim 3 we will determine which protein domains confer KLC functional specificity and will identify and determine the function of adaptors that interact with individual KLCs. Elucidation of the molecular signals regulating neuronal compartmentalization, cargo selection and transport, and axon growth and branching is critical for understanding human developmental disorders, neurodegenerative disorders, and the conditions under which regeneration after axon injury can occur. Our experiments will uncover such mechanisms and thus may help to identify molecular targets for disease treatment.

项目摘要 高度详细和极化的神经元形态的发展需要形成分子不同的 神经元内的隔室以及蛋白质和细胞器的精确亚细胞定位。神经元 隔室化要求诸如驱动蛋白-1之类的电动机将特定的碳传递给 神经元。神经元特别依赖于多样化和高保真货物的运输 偏振和复杂的结构。运输缺陷是多重人类发展和 神经退行性疾病。众所周知，驱动蛋白1可以介导多磅的长距离运输，但 确定开发过程中货物选择和交付特异性的机制差 理解。载能动力蛋白轻链（KLC）驱动蛋白-1（由KLC1-4基因编码）的亚基是 可能涉及特异性，但是他们所调节的发展过程尚不理解。这是一个关键 知识差距，因为人KLC2和KLC4基因的突变引起痉挛性截瘫疾病，很早就 发作。该领域的主要挑战和该项目的长期目标是了解机制 随着神经元在其自然环境中的发展，控制货物运输和本地化 整合多个细胞外提示。我们建立了一个模型，可以在其中图像神经元的动态 在完整的斑马鱼胚胎中发展的感觉神经元内的货物运输。脊椎动物感觉神经元 伸展不同的中央和周围轴突以形成感觉电路。我们发现了独特的功能 神经发育过程中的个别KLC，包括塑造轴突轴的角色，轴突分支，指导 和在开发过程中的维护以及电路功能中的作用。在AIM 1中，我们将确定机制 KLC4调节轴突分支和分隔，以及它运输的细胞器冠。在 AIM 2我们将确定KLC2在轴突开发和货物运输中的功能以及机制 潜在的人类Spoan综合征。在AIM 3中，我们将确定哪些蛋白质结构域赋予KLC功能 特异性并将识别并确定与单个KLC相互作用的适配器的功能。阐明 调节神经元区室化，货物选择和运输的分子信号以及轴突 生长和分支对于理解人类发育障碍，神经退行性至关重要 疾病以及轴突损伤后再生的状况。我们的实验会 发现这种机制，因此可能有助于确定疾病治疗的分子靶标。