Poly (ADP-Ribose) Signalling in Glutamate Excitotoxicity

谷氨酸兴奋毒性中的聚 (ADP-核糖) 信号转导

• 批准号: 6803724
• 负责人: Ted M. Dawson
• 金额: $ 3.69万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6803724
• 关键词: ADP ribosylation; DNA damage; SDS polyacrylamide gel electrophoresis; biological signal transduction; cell death; cerebral ischemia /hypoxia; cysteine endopeptidases; cytotoxicity; enzyme activity; enzyme induction /repression; excitatory aminoacid; free radical oxygen; genetically modified animals; glutamates; immunocytochemistry; laboratory mouse; nerve injury; neurotoxicology; nitric oxide; oxidative stress; pentosyltransferase; stroke; western blottings

Stroke is the third leading cause of death and disability in the United States. Understanding the molecular mechanisms that lead to neuronal injury and cell death may lead to novel therapeutics to treat this devastating disorder. A variety of indices in models of ischemia indicate the involvement of oxygen free radicals, oxidative stress and DNA damage and a potential pivotal role for Poly (ADP-ribose) polymerase-1 (PARP-1) in the pathogenesis of stroke. PARP-1 is a ubiquitous and unique protein which is activated by DNA strand nicks and breaks, which can be induced by neuronal injury. Activation of PARP-1 may lead to the loss of cellular NAD+ and ATP, and the mitochondrial release and translocation of apoptosis inducing factor (AIF) resulting in acute cell injury and death. These events may be crucial in both short- and longer-term deleterious effects of stroke. However, it is unclear how PARP-1 activation leads to AIF release. Is it the NAD+ and ATP loss that accompanies PARP-1 activation that leads to AIF release and cell death or does poly(ADP-ribosyl)ation signal the release of AIF and induce the cell death cascade? Accordingly, this project is designed to explore the molecular mechanisms and consequences of PARP-1 activation following excitotoxic neuronal injury, as well as, the molecular consequences of poly (ADP-ribosyl)ation. In Specific Aim #1 we will evaluate the mechanism by which PARP-1 activation leads to release of AIF from the mitochondria and its translocation to the nucleus following cellular injury. In Specific Aim #2 we will identify PAR polymer binding proteins and investigate their role in cell death induced by PARP-1 activation. In Specific Aim #3 we will determine the role of poly (ADP-ribose) glycohydrolase (PARG) in PARP-1 mediated cell death following reactive oxygen species induced cellular injury and excitotoxic mediated neuronal injury through RNAi approaches and in PARG knockouts. And in Specific Aim #4 we will evaluate the role of overexpression of PARG in PARP-1 mediated cell death following reactive oxygen species induced cellular injury and excitotoxic mediated neuronal. Clarifying and understanding the molecular consequence of PARP-1 activation and the role poly(ADP-ribosyl)ation in the release of AIF and cell death following excitotoxic neuronal injury may yield important insight into the function of PARP-1, poly(ADP-ribosyl)ation, PARG and AIF that may identify novel targets for therapy aimed at preserving neurologic function following excitotoxic neuronal injury and stroke.

中风是美国死亡和残疾的第三大主要原因。了解导致神经元损伤和细胞死亡的分子机制可能导致新的治疗疗法治疗这种毁灭性疾病。缺血模型中的各种指标表明氧自由基，氧化应激和DNA损伤的参与以及Poly（ADP-核糖）聚合酶1（PARP-1）的潜在关键作用在中风的发病机理中。 PARP-1是一个 无处不在的独特蛋白质被DNA链的划痕和断裂激活，可以由神经元损伤诱导。 PARP-1的激活可能导致细胞NAD+和ATP的丧失，线粒体释放和凋亡诱导因子（AIF）的易位导致急性细胞损伤和死亡。这些事件在中风的短期和长期有害影响中可能至关重要。但是，目前尚不清楚PARP-1激活如何导致AIF 发布。是伴随PARP-1激活的NAD+和ATP损失，导致AIF释放和细胞死亡，还是Poly（ADP-核糖基）ation信号AIF的释放并诱导细胞死亡级联反应？因此，该项目旨在探索兴奋性神经元损伤后PARP-1激活的分子机制和后果，以及聚（ADP-核糖基）ation的分子后果。在特定的目标＃1中，我们将评估 PARP-1激活导致AIF从线粒体释放的机制及其在细胞损伤后转移到细胞核的机制。在特定的目标＃2中，我们将鉴定聚合物结合蛋白，并研究其在PARP-1激活引起的细胞死亡中的作用。在特定的目标＃3中，我们将确定在活性氧诱导细胞损伤和通过RNAI方法和PARG敲除中的电氧诱导细胞损伤和兴奋性毒性介导的神经元损伤后，聚（ADP-核糖）糖含量蛋白酶（PARG）在PARP-1介导的细胞死亡中的作用。在特定的目标＃4中，我们将评估PARG在活性氧诱导的细胞损伤和兴奋性介导的神经元后PARP介导的细胞死亡中过表达的作用。澄清和了解PARP-1激活的分子后果以及在兴奋性神经元损伤后释放AIF和细胞死亡中的角色（ADP-核糖基）的角色可能会对PARP-1，PORP-1，POLY（ADP-核糖基）的功能进行重要洞察力，Parg，Parg AIF可能会确定旨在在兴奋性神经元损伤和中风后保留神经系统功能的新型治疗靶标。