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）：一个多世纪以来，我们就知道神经元具有不同的解剖学特征。由于神经元可以有许多树突，并且每个树突可以有许多突触，因此很明显，树突是细胞通讯和功能的重要调节器。自卡哈尔最初观察以来，这些形态特征如何调节细胞功能一直是个谜。在表明枝晶表现出化学区室化方面已经取得了进展。这种区室化的例子是特定树突区域 Ca++ 水平的刺激变化。这些特征表明，树突不是同质的，实际上不仅表现出形态异质性，而且表现出功能异质性。树突生物学的主要问题之一是刺激完整组织中树突的选定区域如何导致细胞反应？这被称为树突整合，其大部分表征使用电生理学和 Ca++ 输出作为树突功能的指标。然而，树突中还发生其他具有 mRNA 靶向和局部翻译的生理过程，这些过程也是树突介导的生理学（包括突触可塑性）的重要调节剂。树突翻译发生在树突长度上称为“热点”的位置，这首先在舒曼和埃伯文实验室同时得到证实。最近，Eberwine 和 Kim 实验室的体外研究证明了高度复杂的树突翻译过程。这些数据以及其他数据强调了分析树突翻译的时间和空间动态以了解突触后反应和树突整合机制的基本需要。迄今为止，大部分翻译工作都利用了培养物中分散的神经元，虽然适用于许多实验问题，但越来越清楚的是，正常微环境中的细胞在细胞生物学（包括 RNA 表达）方面可能与体外对应的细胞在功能上有所不同。本应用中的实验将定义自然微环境中完整神经元树突中多 mRNA 翻译的基本方面。这些数据将首次量化微环境通过调节局部树突蛋白合成的动力学来调节神经元生理学的作用。