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了死亡调节性的概念 基因在缺血性损伤的情况下调节神经元的命运。这 这些研究的假设是生存OS缺血性神经元是 至少部分通过死亡促进剂的表达确定 死亡抑制基因。因此，封锁了表达的影响 死亡启动子基因或死亡抑制基因的过表达或 模仿它们的影响可能会导致中风的新疗法。 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剂或死亡抑制基因的表达 全球性缺血后大鼠脑的增加。克隆这些基因将 允许开发这些基因表达的技术 可以选择性地改变并提供有关机制的见解 缺血性神经元死亡。将解决以下具体目标： 1）识别，完全顺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 在全球缺血死亡之前，在CA1神经元死亡之前诱导。 候选基因包括CPP 32，ICH-2，ICH-2和MCH-2。 2）识别，完全序列并表征大鼠的表达 BCL-2相关的死亡促进剂或死亡抑制剂的脑字体 基因，在全球死亡之前在CA1神经元中诱导的基因 缺血。候选基因包括BAX，BIK和BAK，MCL-L，BFL-1 BAG-L。 3）克隆新颖的基因在缺血大鼠脑中诱导的新基因 减法杂交和差异筛选。候选人死亡 - 启动子基因将通过其在CA1神经元中的表达来鉴定 致死。候选死亡抑制基因将由其识别 在缺血的神经元中的表达，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调节性 基因在全球性缺血后在大鼠大脑中表达，并鉴定 全局缺血诱导的新基因全部表达。效果 这些基因在神经元的存活下将在体外测试 （项目2）和选定的基因将在体内缺血中检查 （项目3）。