Identification of mRNA decay networks in the Drosophila nervous system

果蝇神经系统中 mRNA 衰减网络的鉴定

• 批准号: 8114619
• 负责人: Michael Cleary
• 金额: $ 19万
• 依托单位: UNIVERSITY OF CALIFORNIA, MERCED
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8114619
• 关键词: 4-thiouracil; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Biological; Biological Assay; Biological Process; Cells; Dactinomycin; Data Analyses; Defect; Development; Disease; Drosophila genus; Drosophila melanogaster; Elements; Embryo; Embryonic Nervous System; Equilibrium; Family; Gene Expression; Generations; Genes; Genetic Transcription; Goals; Half-Life; Human; In Vitro; Measurement; Measures; Messenger RNA; Methods; Microarray Analysis; Nervous System Physiology; Nervous system structure; Neurons; Orthologous Gene; Pathway interactions; Pharmaceutical Preparations; Physiologic pulse; Physiological; Population Heterogeneity; Presynaptic Terminals; Process; RNA Decay; RNA Interference; RNA Sequences; RNA Stability; Regulation; Regulatory Element; Reporter; Role; Signal Transduction; Site; Structure; System; Techniques; Testing; Transcript; Uracil phosphoribosyltransferase; Uridine; Validity of Results; Work; base; cell type; computerized tools; genetic regulatory protein; genome-wide; genome-wide analysis; in vivo; knock-down; nerve stem cell; nervous system development; nervous system disorder; neurodevelopment; neurogenesis; new technology; novel; prevent; programs; relating to nervous system; research study; response; uracil analog

DESCRIPTION (provided by applicant): Gene expression levels are determined by the transcription rate of each gene and the decay rate of the resulting mRNA. The balance between these two processes can be altered in response to physiologic conditions, developmental signals, or disease states. Regulation of mRNA decay is particularly important in the nervous system, where the unique structure of neurons requires mRNAs to be selectively stabilized in axon terminals, far from their site of synthesis, and the generation of cellular diversity by neural progenitors requires the programmed decay of mRNAs that regulate proliferation or differentiation. Defective regulation of mRNA decay has been implicated in several human neurological disorders, including Alzheimer's disease and amyotrophic lateral sclerosis. To completely understand the role of mRNA decay in neural development and disease, it will be useful to construct mRNA decay regulatory networks that contain the following information: genome-wide mRNA decay rates in specific cell types under defined conditions, the regulatory proteins that target specific mRNAs for decay, and the mRNA sequence elements that confer coordinate programs of decay. To identify the components of such a network, it is necessary to measure mRNA decay in specific cell types, on a genome-wide scale, under in vivo conditions. Such experiments have historically been difficult, if not impossible, to perform. This project uses a novel technique, known as "TU-tagging", that allows mRNA decay measurements from specific cell types under in vivo conditions. We have recently developed this technique in the model organism, Drosophila melanogaster, and are now well poised to use TU-tagging to identify mRNA decay regulatory networks in the Drosophila nervous system. Our aims are to optimize and validate the accuracy and sensitivity of TU-tagging-based mRNA decay measurements, measure mRNA stability in cells of the Drosophila nervous system, identify the mRNA targets of an mRNA decay regulatory factor encoded by the found in neurons (FNE) gene, and identify cis-regulatory elements among coordinately regulated mRNAs. The specific aims of this work encompass both technical goals and biological goals, reflecting the potential of this project to make significant contributions to the scientific "toolkit" as well as advance the understanding of fundamental biological processes that are relevant to human neural development and disease. PUBLIC HEALTH RELEVANCE: The development and function of the nervous system requires precise control of gene expression and the decay of messenger RNAs (mRNAs) is an essential yet often overlooked control mechanism. Altered regulation of mRNA decay has been implicated in developmental defects and neurological disorders, including Alzheimer's disease and amyotrophic lateral sclerosis. We have developed a novel technology that will allow genome-wide analysis of mRNA decay in neural cells and the construction of comprehensive "blueprints" of mRNA decay pathways that control nervous system development and function.

描述(由申请人提供)： 基因表达水平由每个基因的转录速率和由此产生的mRNA的衰减率决定。这两个过程之间的平衡可以根据生理条件、发育信号或疾病状态而改变。神经元的独特结构要求mRNAs选择性地稳定在远离其合成部位的轴突末端，而神经前体细胞的多样性的产生需要调节增殖或分化的mRNAs的程序性衰变。对mRNA衰退的调节缺陷被认为与几种人类神经疾病有关，包括阿尔茨海默病和肌萎缩侧索硬化症。为了完全了解信使核糖核酸在神经发育和疾病中的作用，构建包含以下信息的信使核糖核酸衰变调控网络将是有用的：特定细胞类型在特定条件下的全基因组信使核糖核酸衰减率，靶向特定信使核糖核酸进行衰变的调节蛋白，以及提供衰变协调程序的信使核糖核酸序列元件。为了确定这样一个网络的组成部分，有必要在体内条件下，在全基因组范围内测量特定细胞类型的mRNA衰变。从历史上看，这样的实验即使不是不可能，也很难进行。该项目使用了一种名为“TU标记”的新技术，它允许在活体条件下测量特定细胞类型的mRNA衰变。我们最近在模式生物--黑腹果蝇中开发了这项技术，现在已经准备好使用TU标签来识别果蝇神经系统中的mRNA衰变调控网络。我们的目标是优化和验证基于TU标签的mRNA衰变测量的准确性和敏感性，测量果蝇神经系统细胞中mRNA的稳定性，识别由在神经元中发现(FNE)基因编码的mRNA衰变调节因子的mRNA靶点，并在协同调控的mRNAs中识别顺式调节元件。这项工作的具体目标包括技术目标和生物学目标，反映了该项目对科学“工具包”作出重大贡献的潜力，以及促进对与人类神经发育和疾病有关的基本生物学过程的理解。 公共卫生相关性： 神经系统的发育和功能需要对基因表达的精确控制，而信使RNA(MRNAs)的衰变是一种重要的但经常被忽视的控制机制。MRNA衰退调节的改变与发育缺陷和神经疾病有关，包括阿尔茨海默病和肌萎缩侧索硬化症。我们已经开发出一种新的技术，它将允许对神经细胞中的mRNA衰退进行全基因组分析，并构建控制神经系统发育和功能的mRNA衰退途径的全面“蓝图”。