ROBOTIZED EXPRESSION PROFILING OF MOUSE AMYGDALA

小鼠杏仁核的自动化表达谱分析

• 批准号: 6392918
• 负责人: JAMES H EBERWINE
• 金额: $ 23.78万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-19 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6392918
• 关键词: amygdala; gene expression; laboratory mouse; method development; microarray technology; molecular biology information system; neurogenetics; single cell analysis

DESCRIPTION (Applicant's abstract): The amygdaloid complex is involved in the "processing" of multiple behaviors including fear, anxiety and learning and memory. There is also evidence that biochemical changes occur in the amygdala as a result of disease including schizophrenia, bipolar disease and cocaine and opiate abuse. The neuronal circuitry connecting various nuclei of the amygdala allowing internuclei communication has also been elucidated for aspects of fear conditioning. Given the central role of the amygdala in processing various types of CNS information an elucidation of the relative levels of mRNA abundance for the mRNAs that are present in the amygdala might provide insight into what neurochemical signals the amygdala responds to. Further since neuronal pathways have been dissected characterization of the expression profile in individual interconnected nuclei may provide information about how one region of the amygdala transfers information to another. We propose to generate molecular fingerprints of neurons within different regions of the amygdala using single cell and single nuclei aRNA amplification and microarray analysis in normal mice. Additionally we will determine how the profiles of these nuclei change in a mouse pharmacological model of bipolar disease involving amygdala response to withdrawal from lithium after chronic lithium treatment. As part of this proposal we propose to robotize the aRNA amplification procedure so that we can generate l000 individual sample templates each partitioned into an individual well in 96 well plates. While we are not proposing to perform 1000 microarray experiments (samples will be pooled as described in the text) having individual cells will permit each to be analyzed separately as required. In an effort to create a National resource of expression profiling data and amplified cells from the amygdala for use by other investigators the normal sample plates will be made available to anyone who requests them on a cost basis. The normal sample expression profiling data will be made available on a public website as soon as the data is confirmed by two array analyses. These data should be useful to investigators studying the amygdala as well as to "bioinformaticists" who are in need of large data sets from multiple samples. Such large data sets should prove useful in the development of pattern search algorithms that are sorely needed to property analyze such complex gene expression data sets.

描述（申请人摘要）：杏仁复合体参与了 “处理”多种行为，包括恐惧、焦虑和学习， 记忆也有证据表明杏仁核会发生生化变化 由于精神分裂症、躁郁症和可卡因等疾病， 滥用鸦片连接杏仁核不同核团的神经回路 允许核间通讯也被阐明了恐惧的方面 条件反射鉴于杏仁核在处理各种信息中的核心作用， CNS信息的类型和mRNA相对水平的阐明 杏仁核中大量存在的mRNA可能会提供 杏仁核对什么样的神经化学信号做出反应。此外，自 神经元通路已经被解剖表征的表达 单个互连核中的剖面可以提供关于如何 杏仁核的一个区域向另一个区域传递信息。我们建议 产生不同区域内神经元的分子指纹， 利用单细胞和单核aRNA扩增和微阵列检测杏仁核 在正常小鼠中进行分析。此外，我们还将确定 在双相情感障碍的小鼠药理学模型中， 涉及杏仁核对慢性锂盐治疗后锂盐戒断的反应 治疗作为该提案的一部分，我们建议将aRNA机器人化， 这样我们就可以产生1000个单独的样本 每个模板分配到96孔板中的单个孔中。虽然我们 不建议进行1000个微阵列实验（样品将 如文中所述合并）具有单独的细胞将允许每个细胞被 根据需要单独分析。为了创造一个国家资源， 来自杏仁核的表达谱数据和扩增的细胞， 其他研究人员正常的样品板将提供给任何人 谁要求他们在成本的基础上。正常样本表达式分析数据 一旦数据得到确认， 两个数组分析这些数据应该对研究人员有用。 杏仁核以及需要大量数据集的“生物信息学家” 从多个样本中。这样的大型数据集应该证明是有用的， 开发模式搜索算法， 分析如此复杂的基因表达数据集。