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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要高度精细和极化的神经元形态的发展需要形成分子上不同的神经元内的区室以及蛋白质和细胞器的精确亚细胞定位。神经元分隔要求诸如kinesin-1之类的马达将特定的货物运送到区域内的不同地点。神经元。神经元特别依赖于多样化和高保真度的货物运输，因为它们的高度极化和复杂的结构。运输缺陷是人类多种发育和发育的基础神经退行性疾病。众所周知，Kinesin-1 可以介导多种货物的长途运输，但在开发过程中确定货物选择和交付的特殊性的机制很差明白了。驱动蛋白-1 的货物结合驱动蛋白轻链 (KLC) 亚基（由 klc1-4 基因编码）是可能涉及特异性，但它们介导的发育过程尚不清楚。这是一个关键的知识差距，因为人类 klc2 和 klc4 基因突变会导致早期的痉挛性截瘫疾病发病。该领域的一个主要挑战和该项目的长期目标是了解其机制当神经元在自然环境中发育时，控制货物运输和定位，它们必须整合多种细胞外线索。我们建立了一个模型，可以在其中对神经元的动态进行成像完整斑马鱼胚胎中正在发育的感觉神经元内的货物运输。脊椎动物感觉神经元延伸不同的中央和外周轴突以形成感觉回路。我们发现了独特的功能神经发育过程中的个体 KLC，包括塑造轴突乔木、轴突分支、引导的作用开发过程中的维护和维护，以及电路功能中的角色。在目标 1 中，我们将确定机制 KLC4 通过它调节轴突分支和区室化，以及它运输的细胞器货物。在目标 2 我们将确定 KLC2 在轴突发育和货物运输中的功能以及机制潜在的人类 SPOAN 综合征。在目标 3 中，我们将确定哪些蛋白质结构域赋予 KLC 功能特异性，并将识别和确定与各个 KLC 相互作用的适配器的功能。说明调节神经元区室化、货物选择和运输以及轴突的分子信号生长和分支对于理解人类发育障碍、神经退行性疾病至关重要疾病，以及轴突损伤后可以发生再生的条件。我们的实验将揭示此类机制，从而可能有助于确定疾病治疗的分子靶点。