Neuroprotective Genes in Ischemic Tolerance

项目1：局灶性脑缺血影响脑内基因表达，而基因表达影响预后。神经保护基因由先兆亚致死性局灶性缺血诱导，从而产生耐受性。缺血耐受也是由全脑缺血和全身内毒素引发的亚致死应激引起的。我们的目标是发现在这些范例中产生耐受性的基因子集。我们以前研究脑缺血基因调控的方法是消减杂交和克隆。在过去的五年里，我们克隆了27个新基因。我们现在将使用微阵列技术来提高我们发现基因的能力，并将努力集中在缺血耐受性的保护性基因上。我们开发了新的小鼠耐受模型，以便使用小鼠基因芯片并利用小鼠基因组的力量。我们的方法是使用几种模型，每种模型都会产生耐受性，但在大脑中具有不同的刺激或区域目标。我们将表征在由先前短暂局灶性缺血或内毒素引发保护的皮质中对局灶性缺血的耐受性，并将表征在由先前短暂全脑缺血保护的CA 1神经元中对全脑缺血的耐受性。多个模型和重复将允许对数据进行复杂的统计分析。 已建立的基因组学核心设施将实现最佳数据管理和分析。太平洋西北国家实验室超级计算机和生物信息学部门的新信息学和数据挖掘合作者为我们的数据分析能力增添了实质性的力量。我们的目标是找到内源性神经保护途径的耐受性建模。为此，我们将鉴定局灶性缺血预处理耐受过程中调控的基因和途径（目的1）;鉴定内毒素引发耐受过程中调控的基因和途径（目的2）;并选择性地鉴定缺血预处理耐受过程中调控的基因和途径。 全脑缺血后CA 1区的脆弱神经元（Aim 3）。在这些耐受性模型中发现的已知和未知功能的基因将被推进到项目二和项目三中，以根据特定的战略计划在体外和体内系统中进行验证。内源性保护系统的阐明应确定中风治疗的新方法。

Project 1: Focal ischemia affects gene expression in brain, and such gene expression affects outcome. Neuroprotective genes are induced by premonitory sublethal focal ischernia resulting in tolerance. Ischemic tolerance also results from sublethal stress caused by global ischemia and by systemic endotoxin priming. Our goal is to discover the subset of genesthat produce tolerance in each of these paradigms. Our previous approach to gene modulation in brain ischenaia was subtractive hybridization and cloning. We cloned 27 new genes over the past five years. We will now use microarray technology to increase our capabilities for gene discovery and focus the efforts on protective genes modeled in ischemic tolerance. We have developed new mouse models of tolerance in order to use murine gene chips and exploit the power of the mouse genome. Our approach is to use several models, each of which results in tolerance but with distinct stimuli or regional targets in brain. We will characterize tolerance to focal ischemia in cortex protected by prior brief focal ischemia or endotoxin priming, and will characterize tolerance to global ischemia in CA 1 neurons protected by prior brief global ischemia. Multiple models and replicates will permit sophisticated statistical analysis of data. Established core facilities for genomics will enable optimal data management and analysis. New informatics and data mining collaborators at Pacific Northwest National Laboratories Supercomputer and Bioinformatics divisions adds substantial strength to our data analysis capabilities. Our goal is to find endogenous pathways of neuroprotection modeled in tolerance. To this end we will identify genes and pathways regulated during tolerance by preconditioning focal ischemia (Aim 1); identify genes and pathways regulated during tolerance by endotoxin priming (Aim 2); and identify genes and pathways regulated during tolerance in the selectively vulnerable neurons of the CA 1 sector following global ischemia (Aim 3). Genes of known and unknown function discovered in these tolerance models will be advanced to projects two and three for validation in in vitro and in vivo systems based on a specific strategic plan.The elucidation of endogenous protective systems should identify novel approaches for stroke treatment.