In vivo translational analysis in neurons

神经元体内翻译分析

• 批准号: 8995218
• 负责人: JAMES H EBERWINE
• 金额: $ 37.5万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-15 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8995218
• 关键词: Area; Biological Assay; Biology; Cell Communication; Cell physiology; Cells; Cellular biology; Characteristics; Chemicals; Chimeric Proteins; Clear Cell; Communication; Complement; Complex; Computer Analysis; Data; Dendrites; Electrophysiology (science); Environment; Exhibits; Functional disorder; Future; Genetic Transcription; Genetic Translation; Goals; Health; Heterogeneity; Hippocampus (Brain); Image; In Vitro; Individual; Intervention; Length; Life; Mediating; Messenger RNA; Monitor; Neurocognitive; Neurons; Output; Physiological; Physiological Processes; Physiology; Population; Preparation; Process; Property; Protein Biosynthesis; Proteins; RNA; RNA analysis; Role; Site; Slice; Specific qualifier value; Synapses; Synaptic plasticity; Therapeutic Intervention; Time; Tissues; Transfection; Translating; Translations; Work; in vivo; insight; mRNA tagging; nervous system disorder; neuropsychiatric disorder; research study; response; transcriptome; transcriptomics

DESCRIPTION (provided by applicant): Neurons have been known to have distinct anatomical specifications for over a century. As neurons can have many dendrites and each dendrite can have many synapses it is clear that dendrites are an important modulator of cellular communication and function. How these morphological features modulate cellular function has been a mystery since the time of Cajal's initial observations. Progress has been made in showing that dendrites exhibit chemical compartmentalization. This compartmentalization is exemplified by stimulated changes in Ca++ levels in specific dendritic areas. These features show that dendrites are not homogeneous and indeed not only exhibit morphological heterogeneity but also functional heterogeneity. One of the dominant questions in dendrite biology is how does stimulation of selected regions of dendrites in the intact tissue result in a cellular response? This has been termed dendritic integration with much of its characterization using electrophysiological and Ca++ outputs as indicators of dendritic function. There are however other physiological processes that occur in dendrites with mRNA targeting and local translation that are also important modulators of dendrite-mediated physiologies including synaptic plasticity. Dendritic translation occurs at sites along the length of the dendrites called "hotspots" first demonstrated simultaneously in the Schuman and Eberwine labs. Recent, in vitro studies from the Eberwine and Kim labs have demonstrated a highly complex dendritic translational process. These data, and those of others, highlight the fundamental need to analyze the temporal and spatial dynamics of translation in dendrites to understand the mechanism of post-synaptic responsiveness and dendritic integration. Much of the translation work to date has utilized dispersed neurons in culture and while appropriate for many experimental questions, it is increasingly clear that cells in their normal microenvironment can be functionally distinct from their in vitro counterparts in their cell biology including RNA expression. Experiments in this application will define the fundamental aspects of multi-mRNA translation in intact dendrites of neurons that are in their natural microenvironment. These data will be the first to quantify the role of the microenvironment in modulating neuronal physiology through modulation of the dynamics of localized dendritic protein synthesis.

描述（由申请人提供）：已知神经元在一个多世纪以来具有独特的解剖学规范。由于神经元可以具有许多树突，并且每个树突可以具有许多突触，因此很明显，树突是细胞通信和功能的重要调节剂。自Cajal初始观察时期以来，这些形态特征如何调节细胞功能一直是一个谜。在表明树突表现出化学隔室化方面取得了进展。在特定树突区域的Ca ++水平的刺激变化中，这种分区化是例证的。这些特征表明，树突不是均匀的，实际上不仅表现出形态异质性，而且表现出功能异质性。树突生物学中的主要问题之一是，完整组织中对树突的选定区域的刺激如何导致细胞反应？这已称为树突状相位，其大部分使用电生理学和Ca ++输出作为树突函数的指标。但是，在树突中还有其他生理过程，具有mRNA靶向和局部翻译，它们也是树突介导的生理学的重要调节剂，包括突触可塑性。树突平移发生在沿树突的长度上，称为“热点”，首先在舒曼和埃伯温实验室中同时证明。最近的Eberwine和Kim Labs的体外研究表明，树突状翻译过程高度复杂。这些数据以及其他数据强调了树突中翻译的时间和空间动力学的基本需求，以了解后突触后反应性和树突构成的机制。迄今为止，大部分翻译工作都在培养中使用了分散的神经元，虽然适合许多实验性问题，但越来越明显的是，其正常微环境中的细胞在功能上可以与其在细胞生物学（包括RNA表达）中的体外对应物有不同。该应用程序中的实验将定义自然微环境中神经元完整树突中多MRNA翻译的基本方面。这些数据将是第一个通过调节局部树突蛋白合成的动力学来量化微环境在调节神经元生理中的作用的数据。