IDENTIFICATION OF DEATH REGULATORY GENES IN ISCHEMIA

项目1：确定脑缺血中的死亡调控基因 越来越多的证据支持死亡监管的概念 在缺血性损伤的背景下，基因调节神经元的命运。这个 这些研究的假设是缺血神经元的存活率是 至少部分地由死亡促进者和 死亡抑制基因。因此，封锁表达的效果 死亡促进基因或死亡抑制基因的过度表达或 模仿它们的效果可能会导致中风的新疗法。 项目1广泛的长期目标是识别和克隆 已知和新的死亡促进或死亡抑制基因，其表达 在全脑缺血后的大鼠脑内的表达增加。克隆这些基因将会 允许开发这些基因表达的技术 可以有选择地改变，并提供对 缺血性神经元死亡。将实现以下具体目标： 1)对大鼠的表达进行鉴定、全序列测定和特征分析 半胱氨酸蛋白酶死亡启动子基因的脑同源基因 在CA1神经元死于全脑缺血之前诱导。 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、全序列测定和鉴定 Bcl2相关死亡促进剂或死亡抑制物的脑内同源物 在CA1神经元死亡前诱导的基因 缺血症。候选基因包括bax、bik、bak、mcl-L、bfl-1、Bak-L。 3)克隆大鼠脑缺血诱导的新基因 消减杂交和差异筛选。候选人死亡- 启动子基因将通过它们在CA1神经元中的表达来鉴定 直到死亡。候选的死亡抑制基因将通过它们的 在缺血后存活的神经元中的表达，例如在CA3或 齿状回。 在这个项目中，我们将确定哪些已知的死亡调节因子 大鼠全脑缺血后相关基因的表达及鉴定 新基因的整体表达是由全脑缺血诱导的。的影响 这些基因对神经元存活的影响将在体外进行测试 (项目2)和选定的基因将在体内进行缺血检测 (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).