Redox Control of Seizure-Induced Neuroinflammation

氧化还原控制癫痫引起的神经炎症

• 批准号: 10302913
• 负责人: MANISHA N PATEL
• 金额: $ 2.34万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10302913
• 关键词: Address; Antioxidants; Astrocytes; Automobile Driving; Brain; Cells; Cysteine; Epilepsy; Genetic Transcription; Glial Fibrillary Acidic Protein; Goals; In Vitro; Inflammation; Inflammation Mediators; Ions; Laboratories; Lead; Link; Messenger RNA; Mitochondria; Modification; Mus; Mutate; Neurons; Nutrient; Oxidation-Reduction; Oxidative Stress; Prosencephalon; Proteins; Rattus; Reactive Oxygen Species; Research; Role; SOD2 gene; Seizures; Signal Transduction; Testing; Up-Regulation; Viral Vector; astrogliosis; base; in vivo; mitochondrial dysfunction; neuroinflammation; novel; parent grant; protein expression; therapeutic target

Mitochondrial Reactive Oxygen Species in Epilepsy-Associated Astrogliosis The parent grant focuses on the interplay between neuroinflammation and redox status. The supplemental research addresses a related but distinct goal to investigate the role of mitochondria in seizure-induced neuroinflammation. Mitochondria integrate the energy requirements of neuronal cells and circuits with nutrients, ions, inflammatory mediators and redox status. Mitochondrial dysfunction, oxidative stress and neuroinflammation have been linked with pathophysiological hyperexcitability associated with epilepsy. Neuroinflammation specifically has been identified as a therapeutic target for epilepsy, however the detailed mechanisms underlying seizure-induced neuroinflammation, including upregulation of astrocyte- specific glial fibrillary acidic protein (GFAP), remain at large. One clue regarding upregulation of GFAP arises from recent studies in our laboratory showing a robust transcriptional upregulation of GFAP in mice lacking the anti-oxidant mitochondrial manganese superoxide dismutase-2 (Sod2) in forebrain neurons, a finding replicated in primary neuronal-glial culture. In contrast, mixed cultures in which neurons were selectively depleted displayed no such upregulation. The goal of this project is to determine if mitochondrial reactive oxygen species (mtROS) generated within neurons can activate neuroinflammation via upregulation of GFAP expression as a result of posttranslational redox modification of a conserved cysteine (Cys294) in GFAP, resulting in long-lasting neuroinflammation. Aim 1 will determine if neuronal mtROS is sufficient to induce GFAP upregulation and astrogliosis in vitro using mixed rat neuronal culture and in vivo using Nex-Cre/Sod2f/f mice microinjected with viral vectors driving GCaMP6f within astrocytes. Aim 2 will determine if this astrogliosis involves redox modification of GFAP protein by mutating Cys294. These studies will reveal novel redox-based therapeutic targets to treat epilepsy.

线粒体活性氧中的癫痫相关星形胶质细胞增生物种 父母赠款专注于神经炎症和氧化还原状态之间的相互作用。这 补充研究针对研究的相关但独特的目标，以调查 线粒体在癫痫发作引起的神经炎症中。线粒体整合能量 具有营养素，离子，炎症介质以及的神经元细胞和电路的要求 氧化还原状态。线粒体功能障碍，氧化应激和神经炎症已已 与与癫痫相关的病理生理过度兴奋性有关。神经炎症 特别是被确定为癫痫的治疗靶标的 癫痫发作引起的神经炎症的基础机制，包括星形胶质细胞的上调 特定的神经胶质原纤维酸性蛋白（GFAP）仍然很大。关于上调的一个线索 GFAP来自我们实验室的最新研究，显示出强大的转录上调 缺乏抗氧化型线粒体超氧化物歧化酶2的小鼠的GFAP （SOD2）在前脑神经元中，这一发现在原发性神经元培养中得到了复制。相比之下， 神经元选择性耗尽的混合培养物没有表现出这种上调。这 该项目的目标是确定是否产生线粒体活性氧（MTROS） 在神经元内可以通过上调GFAP表达来激活神经炎症 GFAP中保守半胱氨酸（CYS294）的翻译后氧化还原修饰 长期神经炎症。 AIM 1将确定神经元MTROS是否足以诱导 使用混合大鼠神经元培养和体内使用GFAP上调和星形胶质细胞增多 Nex-cre/Sod2f/f小鼠在星形胶质细胞中驱动GCAMP6F的病毒载体微注射。目标2 将确定这种星形胶质细胞增多是否涉及通过突变进行GFAP蛋白的氧化还原修饰 CYS294。这些研究将揭示基于氧化还原的新型治疗靶标以治疗癫痫。