Redox Control of Seizure-Induced Neuroinflammation

癫痫相关性星形胶质细胞增生的线粒体活性氧 父母资助的重点是神经炎症和氧化还原状态之间的相互作用。的 补充研究解决了一个相关但不同的目标，调查的作用， 线粒体在糖尿病诱导的神经炎症中的作用。线粒体整合能量 神经元细胞和回路对营养素、离子、炎症介质和 氧化还原状态线粒体功能障碍、氧化应激和神经炎症已被 与癫痫相关的病理生理性过度兴奋有关。神经炎症 已经被确定为癫痫的治疗靶点，然而， 神经炎性反应的潜在机制，包括星形胶质细胞的上调， 特异性胶质细胞酸性蛋白（GFAP）仍然逍遥法外。一个关于上调 GFAP源于我们实验室最近的研究，显示出强大的转录上调 缺乏抗氧化剂线粒体锰超氧化物歧化酶-2的小鼠中GFAP的表达 （Sod 2）在前脑神经元中，在原代神经胶质细胞培养中复制的发现。与此相反， 其中神经元被选择性耗尽的混合培养物没有显示出这样的上调。的 该项目的目标是确定线粒体活性氧（mtROS）是否产生 在神经元内，可以通过上调GFAP表达来激活神经炎症， GFAP中保守的半胱氨酸（Cys 294）的翻译后氧化还原修饰，导致 长期的神经炎症目的1将确定神经元mtROS是否足以诱导 在体外使用混合大鼠神经元培养和在体内使用GFAP上调和星形胶质细胞增生 用病毒载体显微注射Nex-Cre/Sod 2f/f小鼠，所述病毒载体在星形胶质细胞内驱动GCaMP 6 f。目的2 将确定这种星形胶质细胞增生是否涉及GFAP蛋白的氧化还原修饰， Cys294这些研究将揭示治疗癫痫的新的基于氧化还原的治疗靶点。

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.