ROBOTIZED EXPRESSION PROFILING OF MOUSE AMYGDALA

描述(申请人摘要)：杏仁复合体参与了 对包括恐惧、焦虑和学习在内的多种行为进行“加工” 记忆。也有证据表明杏仁核发生了生化变化。 由于精神分裂症、躁郁症和可卡因等疾病 鸦片类药物滥用。连接杏仁核不同核团的神经元回路 对于恐惧的各个方面，也已经阐明了允许核间交流 条件反射。鉴于杏仁核在处理各种 中枢神经系统的信息类型及信使核糖核酸相对水平的阐明 杏仁核中存在的丰富的mRNAs可能提供洞察力 杏仁核对什么神经化学信号做出反应。自那以后 神经元通路已被解剖表征的表达 单个相互连接的原子核的轮廓可以提供关于如何 杏仁核的一个区域向另一个区域传递信息。我们建议 产生不同区域神经元的分子指纹 杏仁核单细胞和单核RNA扩增及微阵列技术 在正常小鼠身上进行分析。此外，我们还将确定配置文件如何 在双相情感障碍的小鼠药理模型中这些核团的变化 慢性锂中毒后杏仁核对戒断锂的反应 治疗。作为这项提议的一部分，我们提议将阿尔纳机器人 扩增程序，以便我们可以产生1000个单独的样本 每个模板被划分到96个井板中的一个单独的井中。当我们 不建议进行1000个微阵列实验(样本将 如正文中所描述的汇集)具有单独的单元格将允许每个 根据需要分别进行分析。在努力创建一个国家资源的过程中 来自杏仁核的表达谱数据和扩增细胞用于 其他调查人员将向任何人提供正常的样本模板 他们是在成本的基础上提出要求的。正态样本表达谱数据 一旦数据得到确认，将立即在公共网站上提供 两个数组分析。这些数据应该对研究 杏仁核以及需要大数据集的“生物信息学家” 从多个样本中提取。如此庞大的数据集应该会被证明在 开发属性迫切需要的模式搜索算法 分析这种复杂的基因表达数据集。

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.