CNS transcription modulation in peripheral inflammation

描述(申请人提供)：这一假说生成研究提案的目标是研究与疾病相关的分子机制 中枢神经系统(CNS)对全身炎症的反应。中环 对全身炎症的反应将通过DNA微阵列进行监测 技术，这允许同时监测的表达 成千上万的基因。我们提议的研究将系统地评估几个 将为在研究中使用微阵列提供指导的参数 整个中枢神经系统组织的分离区域，并将包括广泛的 用实时荧光定量聚合酶链式反应对研究结果进行验证。我们将评估以下参数 批内(玻片间)和批间变异系数，以及 包括脑区特异的和炎症的cdna对照。我们会 评估使用以下方法获得的数据：(1)三种组织解剖方法，(2)两种 大脑区域(下丘脑和海马体)，以及(3)特定的转基因 用我们选择的范例显示有意义的生物学结果的模型。这个 对这些参数的验证和我们数据分析的含义将 作为一种综合性的技术，在评价微阵列方面是有用的 可能发生的中枢神经系统转录变化的调查 完成当前的基因组计划。为了研究CNS基因转录，我们将 使用全身炎症反应综合征(SIRS或脓毒症)的啮齿动物模型 由外周(腹膜)脂多糖(LPS)注射引起。 这是一种临床相关的炎症模型，可导致急性 中枢神经系统中多种基因的转录。要研究的领域， 下丘脑和海马体是临床上调节脑电活动的部位。 与炎症相关的神经内分泌反应。此外，建议的 研究将产生与转录相关的新的有价值的数据 在定义信号通路机制中起重要作用的调节 炎症状态下的中枢神经系统。拟议的研究将产生新的数据 这将是我们产生新的可检验假说所必需的，这将 为未来的R01应用程序提供框架。

DESCRIPTION (provided by applicant): The goal of this hypothesis-generating research proposal is to investigate the molecular mechanisms involved in the central nervous system (CNS) response to systemic inflammation. The central response to systemic inflammation will be monitored by DNA microarray technology, which allows the simultaneous monitoring of the expression of thousands of genes. Our proposed studies will systematically assess several parameters that will provide guidance for the use of microarrays in the study of dissected discrete regions of whole CNS tissue and will include extensive validation of the findings by real-time PCR. We will assess parameters such as coefficients of intraassay (interslide) and interassay variation, and the inclusion of brain region specific and inflammation cDNA controls. We will evaluate data obtained using: (1) three methods of tissue dissection, (2) two brain regions (hypothalamus and hippocampus), and (3) a specific transgenic model that shows meaningful biological outcome with our chosen paradigm. The validation of these parameters and the implications of our data analyses will be useful in the evaluation of microarray as a technique for comprehensive surveys of CNS transcriptional changes that will be likely to occur upon the completion of current genomic projects. To study CNS gene transcription we will use a rodent model of systemic inflammation response syndrome (SIRS or Sepsis) caused by peripheral (intraperitoneal) lipopolysaccharide (LPS) administration. This is a clinically relevant model of inflammation that induces acute transcription of multiple genes in the CNS. The areas to be studied, hypothalamus and hippocampus, are the sites for the regulation of clinically relevant neuroendocrine responses to inflammation. In addition, the proposed studies will result in new and valuable data related to transcription regulation that are important in defining signaling pathway mechanisms in the CNS during states of inflammation. The proposed studies will result in new data that will be needed for us to generate novel testable hypotheses, which will provide the framework for future R0l applications.