A bioinformatics approach to pannexin localization and interaction in the brain

大脑中pannexin定位和相互作用的生物信息学方法

• 批准号: 7750696
• 负责人: Angela C Cone
• 金额: $ 4.52万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7750696
• 关键词: Address; Antibodies; Architecture; Area; Arts; Bioinformatics; Brain; Brain Mapping; California; Cell Culture Techniques; Cell physiology; Cells; Complement; Complex; Computer Simulation; Connexins; Cytoskeletal Proteins; Data; Data Collection; Databases; Deposition; Electron Microscopy; Fluorescence; Goals; Heart; Hot Spot; Human Genome; Image; Imaging Techniques; Invertebrates; Laboratories; Light; Maps; Membrane; Mentors; Methods; Microscopic; Microscopy; Mining; Modeling; Molecular; Mus; Nervous system structure; Organ; Proteins; Purinoceptor; Research; Resolution; Rodent; Signal Transduction Pathway; Structure; Synapses; System; Systems Integration; Tertiary Protein Structure; Testing; Tissues; Universities; base; computerized tools; data mining; data sharing; gap junction channel; glycosylation; information processing; intercellular communication; millimeter; neuroinformatics; protein expression; protein structure; receptor; research study; systems research; tool; unpublished works

DESCRIPTION (provided by applicant): Recent in silico searching of the human genome yielded three related proteins - the pannexins (Panxl, Panx2 and Panx3), originally speculated to function as gap junction channels because of the their connexin- like protein domain organization. Although connexins are the most widely studied intercellular communication channels, non-connexin gap junction channels have been discovered experimentally in invertebrates (innexins) and pannexins are more similar to innexins than connexins. The tissue structure analyses proposed in this research will investigate, using large-scale fluorescence light microscopic montage imaging, Panxl and Panx2 expression in the rodent brain to determine areas with high levels of expression and then use data sharing, mining and neuroinformatic tools to see whether pannexins co- localize with other proteins as interaction partners or indicate that they are part of a larger cellular complex. This project aims to integrate experimental imaging with computational analyses in order to obtain quantitative and interdisciplinary information about pannexin expression/function. This proposal utilizes a data-intensive mosaic imaging technique for large-scale mapping of the mouse brain combined with neuroinformatics, data mining, and sharing to help integrate and synthesize data for the broader impact of modeling the complex mammalian brain. Specifically, the goals of my research are: (1) To apply our recently developed antibody tool kit to image whole mouse brain at high light microscopic resolution where both Panxl and Panx2 are highly expressed, (2) To deposit and annotate the brain maps in public databases for data sharing, (3) To use the brain maps in conjugation with neuroinformatics and data mining computational tools to determine if there are "hot spots" for Panxl and Panx2 localization in the mouse brain correlated with high expression of other proteins such as connexins or purinergic receptors, (4) To correlate membrane versus intracellular localizations of endogenous Panxl and Panx2 in tissue to complement ex vivo studies of exogenously expressed pannexins in cell culture. Relevance: This is part of a multi-scale coordinated approach with directions in molecular, cellular and organ level imaging of Panxl and Panx2. This multilevel approach addresses the particular challenge for nervous system research to bridge the dimensional range from tissues to molecules, a range encompassing cellular networks, dendritic and axonal architectures, synaptic connectivity, glial interactions and macromolecular constituents. These structures, and the proteins within them, represent the heart of information processing in the nervous system; the integration, synthesis, and sharing of this protein expression data is central to our understanding of brain function.

描述（由申请人提供）：最新的人类基因组硅搜索产生了三种相关的蛋白质 - Pannexins（Panxl，Panx2和Panx3），最初推测是由于其类似于结的蛋白蛋白领域的组织，因此推测起了间隙连接通道的作用。尽管连接蛋白是研究最广泛的细胞间通信通道，但在无脊椎动物（innexins）实验中发现了非连续性间隙连接通道，而Pannexins比connexins比连接蛋白更相似。这项研究中提出的组织结构分析将使用啮齿动物大脑中的大规模荧光微观蒙太奇成像，PANXL和PANX2表达表达，以确定具有高表达的区域，然后使用数据共享，采矿和神经信息性工具，以查看与其他蛋白质相互作用的部分是否与其他蛋白质进行了构图，它们是否具有较大的细胞，它们是较大的细胞构造的，它们是一个更大的细胞构造的，它们是一个较大的细胞的一部分。该项目旨在将实验成像与计算分析集成，以获取有关Pannexin表达/功能的定量和跨学科信息。该建议利用数据密集型的镶嵌成像技术来大规模映射小鼠脑，并结合神经信息，数据挖掘和共享，以帮助整合和合成数据，从而对复杂的哺乳动物大脑进行更广泛的影响。具体而言，我的研究的目标是：（1）应用我们最近开发的抗体工具套件以高光显微镜分辨率对整个鼠标大脑进行图像，其中PANXL和PANX2都高度表达，（2）在公共数据库中存放和注释大脑图以进行数据共享，（3）在与脑图中使用脑图，以确定与neuroining和数据相结合的脑图，以确定'neuroining和Data into''小鼠脑中的PANX2定位与其他蛋白质（例如连接蛋白或嘌呤能受体）的高表达相关，（4）将内源性panxl和Panx2在组织中的膜与细胞内局部局部相关联，以补充细胞培养中源自表达的泛蛋白在细胞培养中的体内研究。相关性：这是多尺度协调方法的一部分，该方法在PANXL和PANX2的分子，细胞和器官水平成像方面。这种多级方法解决了神经系统研究的特殊挑战，即桥接从组织到分子的尺寸范围，包括细胞网络，树突状和轴突结构，突触连通性，神经胶质相互作用和大分子成分。这些结构以及其中的蛋白质代表了神经系统中信息处理的核心。该蛋白质表达数据的整合，合成和共享对于我们对大脑功能的理解至关重要。