IDENTIFICATION OF DEATH REGULATORY GENES IN ISCHEMIA

项目1：缺血死亡调控基因的鉴定 越来越多的证据支持这一概念，即死亡调节 基因调节缺血性损伤情况下神经元的命运。的 这些研究的假设是，缺血神经元的存活是 至少部分地由死亡启动子的表达决定， 死亡抑制基因因此，阻断表达的影响， 死亡启动子基因或死亡抑制基因的过表达，或 模仿它们的作用可能会导致中风的新疗法。 项目1的广泛的长期目标是识别和克隆 已知的和新的死亡启动子或死亡抑制基因，其表达 在全脑缺血后大鼠脑中增加。克隆这些基因将 允许开发技术，通过这些技术， 可能会被选择性地改变，并提供深入了解的机制， 缺血性神经元死亡。将处理以下具体目标： 1)识别、完全测序和表征大鼠的表达 半胱氨酸蛋白酶死亡启动子基因的脑同源物， 在CA 1神经元因全脑缺血死亡之前诱导。 候选基因包括cpp 32，ich-2，ich-2和mch-2。 2)对大鼠的表达进行鉴定、完全测序和表征 bcl-2相关死亡启动子或死亡抑制子的脑同源物 这些基因在CA 1神经元死亡前被诱导， 缺血候选基因包括Bax、bik和巴克、mcl-1、bfl-1、bag-l。 3)克隆脑缺血诱导的新基因 差减杂交和差异筛选。候选人死亡- 启动子基因将通过它们在CA 1神经元中的表达来鉴定， 至死候选的死亡抑制基因将通过其 在缺血后存活的神经元中表达，例如在CA 3或CA 4中的那些。 齿状回 在这个项目中，我们将确定哪些已知的死亡监管 基因在大鼠全脑缺血后表达，并鉴定 全脑缺血诱导新基因全表达。的影响 将在体外测试这些基因对神经元存活的影响 （项目2）和选定的基因将在体内缺血检查 （项目3）。

Project 1: Identification of Death-Regulatory Genes in Ischemia A growing body of evidences supports the concept that death regulatory genes modulate the fate of neurons in the setting of ischemic injury. The hypothesis of these studies is that survival os ischemic neurons is determined, at least in part, by the expression of death-promoter and death-suppressor genes. Thus, blockade of the expression of the effects of death promoter genes or overexpression of death suppressor genes or mimicking their effects could result in novel therapies for stroke. The broad long-term objectives of project 1 are to identify and clone known and novel death-promoter or death-suppressor genes whose expression is increased in rat brain after global ischemia. Cloning these genes will allow for the development of techniques by which expression of these genes may be selectively altered and provide insights into the mechanism of ischemic neuronal death. The following specific aims will be addressed: 1) Identify, fully sequence, and characterize the expression of the rat brain homologues of cysteine protease death promoter genes that are induced in CA1 neurons prior to their death from global ischemia. Candidate genes include cpp 32, ich-2, ich-2 and mch-2. 2) Identify, fully sequence and characterize the expression of the rat brain homologues of bcl-2-related death-promoter or death-suppressor genes, that are induced in CA1 neurons prior to their death from global ischemia. Candidate genes include Bax, bik, and bak, mcl-l, bfl-1 bag-l. 3) Clone novel genes induced in ischemic rat brain identified by subtractive hybridization and differential screening. Candidate death- promoter genes will be identified by their expression in CA1 neurons prior to death. Candidate death-suppressor genes will be identified by their expression in neurons that survive ischemia, such as those in CA3 or dentate gyrus. In this project we will determine which of the known death-regulatory genes are expressed in rat brain following global ischemia and identify novel genes whole expression is induced by global ischemia. The effect of these genes upon the survival of the neuron will be tested in vitro (project 2) and selected genes will be examined in ischemia in vivo (project 3).