Subcellular Proteomic Investigation of Projection Neuron Growth Cones in Developing Mouse Cortex

发育中的小鼠皮层投射神经元生长锥的亚细胞蛋白质组学研究

基本信息

批准号：
10750664
负责人：
Dustin Eliot Tillman
金额：
$ 3.72万
依托单位：
HARVARD UNIVERSITY
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 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10750664
关键词：
Adult Axon Behavior Behavioral Cell Nucleus Cells Cerebral cortex Chemicals Cognition Cognitive Complement Complex Contralateral Cues Degradation Pathway Development Diameter Disease Environment Esthesia Future Genes Genetic Transcription Growth Cones Hour In Vitro Intellectual functioning disability Investigation Link Maintenance Maps Mass Spectrum Analysis Molecular Motor Movement Mus Nature Neurodegenerative Disorders Neurodevelopmental Disorder Neurons Nuclear Parents Pathway interactions Play Process Protein Biosynthesis Proteins Proteome Proteomics RNA RNA Splicing Regulation Research Role Sensory Signal Transduction Social Behavior Synapses Translational Regulation Variant Work Zinc Fingers autism spectrum disorder axon growth axon guidance axonal pathfinding candidate selection causal variant combinatorial extracellular genetic approach genetic manipulation in vivo nervous system disorder neural circuit neurodevelopment neuron development neuronal circuitry neuronal growth novel protein degradation protein function proteostasis response skills subcellular targeting synaptogenesis therapeutic target transcription factor transcriptome

项目摘要

Project Summary/Abstract During development of the cerebral cortex, cortical projection neuron subtypes extend axons to innervate distinct targets located at great distances (103-105 cell body diameters) from their nucleus-containing somata. This precise navigation and circuit development is regulated by growth cones (GCs): subcellular compartments at tips of growing axons that rapidly integrate extracellular signals to control development of neural circuits, then mature into synapses. Since GCs respond rapidly to chemical cues, while hours/days are required to transport molecules between GCs and their parent somata, GCs highly likely function semi-autonomously. Prior work has also revealed that local protein synthesis and degradation pathways in GCs are required for responses to some directional cues. However, function of most GC proteins in axon guidance and circuit development have not been identified, and even less is known about subtype-specific roles of GC proteins. This proposal combines recently developed subtype-specific GC purification with ultra-low-input proteomics to 1) investigate subtype-specific GC proteomes at distinct developmental stages (pre- and post-midline crossing) to identify and functionally investigate proteins with stage-specific roles, and 2) investigate subtype-specific proteomes of dysfunctional GCs to identify and functionally investigate dysregulated proteins with critical roles in precise circuit wiring. These rigorous in vivo investigations will deepen understanding of subtype-, and stage-specific mechanisms regulating subcellular proteostasis in GCs, and how these processes control axon pathfinding and formation of synaptic circuitry in cortical projection neuron subtypes. The intersection of circuit formation and subcellular proteostasis has substantial

项目概要/摘要在大脑皮层发育过程中，皮层投射神经元亚型延伸轴突以进行神经支配不同的目标位于距其含核体体较远的距离（103-105个细胞体直径）。这种精确的导航和电路发育受到生长锥 (GC) 的调节：亚细胞区室在生长轴突的尖端，快速整合细胞外信号来控制神经回路的发育，然后成熟为突触。由于 GC 对化学信号反应迅速，而运输需要数小时/数天 GC 与其母体体细胞之间存在分子，因此 GC 很可能半自主地发挥功能。之前的工作有还揭示了 GC 中的局部蛋白质合成和降解途径是响应某些方向性提示。然而，大多数 GC 蛋白在轴突引导和电路发育中的功能尚未得到证实。人们对 GC 蛋白的亚型特异性作用知之甚少。该提案结合了最近开发了具有超低输入蛋白质组学的亚型特异性 GC 纯化，以 1) 研究亚型特异性 GC 不同发育阶段（中线交叉前和中线交叉后）的蛋白质组，以识别和功能研究具有特定阶段作用的蛋白质，2) 研究功能失调的 GC 的亚型特异性蛋白质组识别和功能研究在精确电路布线中发挥关键作用的失调蛋白。这些严格的体内研究将加深对亚型和阶段特异性机制的理解调节 GC 中的亚细胞蛋白质稳态，以及这些过程如何控制轴突寻路和形成皮质投射神经元亚型中的突触回路。电路形成与亚细胞的交叉点蛋白质稳态具有显着的