Role of prohibitin in ischemic brain injury

抑制素在缺血性脑损伤中的作用

• 批准号: 10201370
• 负责人: Ping Zhou
• 金额: $ 49.92万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201370
• 关键词: Address; Affect; Astrocytes; Bioenergetics; Biological Assay; Brain; Brain Injuries; Calcium; Cell Culture Techniques; Cell Survival; Cells; Cellular Assay; Cerebrum; Coculture Techniques; Complement; Complex; Cysteine; Disease; Embryo; Environment; Funding; Gel; Genetic; Genus Hippocampus; Glucose; Ischemia; Ischemic Brain Injury; Ischemic Neuronal Injury; Ischemic Preconditioning; Knock-in; Knock-in Mouse; Knock-out; Knockout Mice; Life; Link; Maintenance; Mediating; Mitochondria; Mitochondrial Proteins; Modeling; Molecular; Molecular Weight; Mouse Protein; Mus; Mutant Strains Mice; Mutate; Mutation; Neuroglia; Neurologic; Neurons; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthetase Inhibitor; Open Reading Frames; Oxidative Phosphorylation; Oxygen; Patients; Play; Process; Production; Protein S; Proteins; Reactive Oxygen Species; Regulation; Respiratory Chain; Role; S-Nitrosoglutathione; S-Nitrosothiols; Somatic Mutation; Stress; Stroke; Subfamily lentivirinae; System; Testing; Transgenic Mice; Up-Regulation; Work; artery occlusion; base; cell type; conditional knockout; deprivation; in vivo; ischemic injury; mitochondrial dysfunction; mutant; nervous system disorder; neuron loss; neuronal survival; neuroprotection; novel; overexpression; prevent; prohibitin; protective effect; stroke risk; targeted treatment; treatment strategy

ABSTRACT The mitochondrial protein Prohibitin (PHB) is essential for life. Its importance to cellular activities is attested by the fact that deletion of PHB is embryonic lethal in mice and that to date no mutation has been found in the coding region of PHB in any disease conditions, indicating that PHB integrity is essential and that somatic mutation is detrimental. PHB has been shown to be critical for mitochondrial function in stress situations, as it is upregulated in ischemic preconditioning (IPC). In the previous funding period, using neuron specific PHB transgenic mice, we demonstrated that selective neuronal PHB expression leads to remarkable neuroprotection against middle cerebral arterial occlusion (MCAO) induced brain injury. However, how this important protein is functionally regulated in IPC, as well as how it is dysregulated in other neurological conditions, remain surprisingly unknown. In exploring the mechanisms of PHB regulation, we discovered that nitric oxide (NO) is required for both IPC and PHB upregulation. Therefore, we investigated the interaction between NO and PHB and found that NO modifies PHB post-translationally, through protein s-nitrosylation. In this renewal application, we propose to study the effects of PHB S- nitrosylation and the mechanisms underlying functional regulation of PHB by NO. Our central hypothesis is that PHB nitrosylation is critical for its neuroprotective function and that disturbances of PHB nitrosylation is detrimental. We will use a mutant knockin mouse, in which the sole cysteine residue of PHB protein is mutated so that PHB cannot be nitrosylated, and a PHB neuronal knockout mouse, in which PHB deletion is complemented by AAV expressing wild type or non-nitrosylated C69S mutant to analyze the mechanisms of NO regulation and the effects of loss of PHB nitrosylation on PHB function, in IPC. Three specific aims will systematically test the hypothesis. The results of the proposed studies will reveal a previously undescribed regulatory mechanism of PHB and could benefit patients at risk of stroke and other neurological conditions.

抽象的 线粒体蛋白禁止素（PHB）对生命至关重要。它对细胞活动的重要性 PHB的缺失在小鼠中是胚胎致死的事实证明，迄今为止没有突变 在任何疾病条件下，在PHB的编码区域都发现了PHB完整性 是必不可少的，体​​细胞突变是有害的。 PHB已被证明对 在应力情况下，线粒体功能在缺血性预处理（IPC）中被上调。 在上一个资金期间，使用神经元特异性PHB转基因小鼠，我们证明了 选择性神经元PHB表达导致对大脑中间的神经保护显着 动脉阻塞（MCAO）诱导脑损伤。但是，这种重要的蛋白质在功能上如何 在IPC中受到调节，以及在其他神经系统条件下如何失调 令人惊讶的是未知。在探索PHB调节的机制时，我们发现了一氮 IPC和PHB上调都需要氧化物（NO）。因此，我们调查了 NO和PHB之间的相互作用，发现No在翻译后通过 蛋白质S-亚硝基化。在此续签应用中，我们建议研究PHB S-的影响 硝基化和PHB功能调节的基础机制。我们的中心 假设是PHB亚硝基化对其神经保护功能至关重要，并且干扰 PHB亚硝基化是有害的。我们将使用一种突变的敲蛋白鼠标，其中鞋底 PHB蛋白的半胱氨酸残基被突变，因此PHB不能被硝基基化，并且PHB 神经元基因敲除鼠标，其中PHB缺失与表达野生型的AAV相辅相成 或非硝基化的C69S突变体，以分析无调节的机制和 IPC中PHB功能上PHB硝基化的丧失。三个具体目标将系统地测试 假设。拟议的研究结果将揭示先前未描述的调节 PHB的机制，可以使患有中风和其他神经系统疾病风险的患者受益。