IDENTIFICATION OF DEATH REGULATORY GENES IN ISCHEMIA

缺血中死亡调节基因的鉴定

• 批准号: 6494878
• 负责人: STEVEN H GRAHAM
• 金额: $ 38.25万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6494878
• 关键词: antisense nucleic acid; apoptosis; cerebral ischemia /hypoxia; complementary DNA; gene induction /repression; laboratory rat; neural degeneration; neurogenetics; neurons; oligonucleotides; transcription factor

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).

项目1：缺血中死亡调节基因的鉴定 越来越多的证据支持死亡监管这一概念 基因在缺血性损伤的情况下调节神经元的命运。这 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剂和 死亡抑制基因。因此，阻断效应的表达 死亡促进基因或死亡抑制基因的过度表达或 模仿它们的作用可能会产生新的中风疗法。 项目 1 的广泛长期目标是识别和克隆 已知和新颖的死亡促进基因或死亡抑制基因，其表达 大鼠大脑整体缺血后增加。克隆这些基因将 允许开发表达这些基因的技术 可以有选择地改变并提供对机制的见解 缺血性神经元死亡。将实现以下具体目标： 1) 鉴定、完全测序并表征大鼠的表达 半胱氨酸蛋白酶死亡启动子基因的脑同源物 在 CA1 神经元因整体缺血死亡之前，它们会被诱导。 候选基因包括cpp 32、ich-2、ich-2 和mch-2。 2) 鉴定、完全测序并表征大鼠的表达 bcl-2相关死亡启动子或死亡抑制子的脑同源物 基因，这些基因在 CA1 神经元因全局死亡之前被诱导 缺血。候选基因包括 Bax、bik 和 bak、mcl-l、bfl-1 bag-l。 3) 克隆在缺血大鼠大脑中诱导的新基因 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 神经元中的表达来识别 至死。候选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 CA3 或 齿状回。 在这个项目中，我们将确定哪些已知的死亡监管 基因在大鼠大脑中在整体缺血后表达并识别 新基因的整体表达是由整体缺血诱导的。的效果 这些神经元存活后的基因将在体外进行测试 （项目2）和选定的基因将在体内缺血中进行检查 （项目3）。