Distal mRNA localization and translation in neural stem cells of the developing brain

发育中大脑的神经干细胞中的远端 mRNA 定位和翻译

基本信息

批准号：
10435490
负责人：
Debra Silver
金额：
$ 35.66万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10435490
关键词：
Active Biological Transport Address Binding Binding Proteins Biological Assay Biology Brain Cell Separation Cells Cytoskeleton Data Development Diagnostic Distal Elements Embryo Embryonic Structures Etiology Exhibits FMR1 Fragile X Syndrome Functional disorder Gene Expression Gene Expression Regulation Generations Genes Human Image Imaging Device In Situ In Vitro Kinetics Lead Measures Meninges Messenger RNA Methods Microcephaly Molecular Monitor Mus Nervous system structure Neurodevelopmental Disorder Neuroglia Neurons Pathway interactions Pattern Post-Transcriptional Regulation Process Proteins Proteomics RNA RNA Binding RNA metabolism RNA-Binding Proteins Radial Regulation Reporter Role Sampling Signal Transduction Structure Synapses Testing Therapeutic Tissues Transcript Translating Translational Regulation Translations Tretinoin autism spectrum disorder axon guidance genomic tools human fetal brain imaging approach in vivo in vivo Model infancy insight migration molecular imaging mutant nerve stem cell nervous system disorder neuroregulation novel protein function relating to nervous system transcriptome

项目摘要

Abstract mRNA localization and local translation are conserved mechanisms enabling spatial and temporal control of gene expression. These processes are particularly important in the nervous system, where local regulation of gene expression influences axon guidance and synaptic activity. Much of our understanding of mRNA localization and local translation has relied upon in vitro studies. We recently established a novel tractable in vivo paradigm for investigating local gene regulation in vivo in the developing nervous system. Specifically we found that mRNA is sub-cellularly localized and locally translated in basal structures of embryonic radial glial cells (RGCs), called endfeet. These basal structures are essential for tissue integrity and neuronal migration, thus impacting the central function of RGCs in neuron generation. We developed live imaging approaches to discover that mRNA can be actively transported in RGCs and locally translated in basal endfeet. We also developed novel methods to isolate RGC endfeet and used these to discover a local transcriptome in endfeet, bound by the RNA binding protein, FMRP. Our studies lead us to hypothesize that RNA localization and local translation are intrinsically and extrinsically regulated in RGC endfeet to influence signaling and the cytoskeleton. In this proposal we aim to implement these new molecular imaging and genomic tools towards: (1) discovering local transcriptomes of RGC endfeet across development, (2) determining intrinsic and extrinsic regulation of translation in RGC endfeet, and (3) defining functions for RNA localization in RGC endfeet. Successfully completed, we will have significantly advanced our understanding of mRNA localization and translation in the developing nervous system, by studying these processes in an in vivo model.

抽象的 mRNA 定位和局部翻译是保守机制，能够实现空间和时间控制基因表达。这些过程在神经系统中特别重要，其中局部调节基因表达影响轴突引导和突触活动。我们对 mRNA 的大部分了解本地化和本地翻译依赖于体外研究。我们最近建立了一个新颖的易处理的用于研究发育中神经系统体内局部基因调控的体内范例。具体来说我们发现 mRNA 是亚细胞定位的，并在胚胎放射状胶质细胞的基底结构中进行局部翻译细胞（RGC），称为端脚。这些基底结构对于组织完整性和神经元迁移至关重要，从而影响 RGC 在神经元生成中的中心功能。我们开发了实时成像方法发现 mRNA 可以在 RGC 中主动转运并在基端足中进行局部翻译。我们也开发了分离 RGC endfeet 的新方法，并利用这些方法发现了 endfeet 中的局部转录组，被 RNA 结合蛋白 FMRP 结合。我们的研究使我们假设 RNA 定位和局部 RGC 末端脚的翻译受到内在和外在的调节，以影响信号传导和细胞骨架。在本提案中，我们的目标是实施这些新的分子成像和基因组工具： (1) 发现 RGC 端足在整个发育过程中的局部转录组，(2) 确定内在和外在的转录组 RGC endfeet 中翻译的调节，以及 (3) 定义 RGC endfeet 中 RNA 定位的功能。如果成功完成，我们将显着提高我们对 mRNA 定位和通过在体内模型中研究这些过程，在发育中的神经系统中进行翻译。