Javits Award Final Three Years

项目摘要：中枢神经系统(CNS)的病毒感染与 癫痫发作的风险、癫痫持续状态(SE)和慢性癫痫的发展。我们的协作小组已经 开发了第一个病毒诱导癫痫的动物模型。小鼠(C57BL/6)接受脑内注射 注射泰勒氏小鼠脑脊髓炎病毒(TMEV)显示几种急性自发性癫痫 感染几天后，在最初的感染中存活下来，并继续发展为自发性反复发作。 此外，我们上一次获奖期的数据使用了带有各种细胞因子或细胞因子的C57BL/6小鼠 受体被敲除表明，在肿瘤坏死因子α系统的操纵可以显著改变病理。 观察到TMEV注射后的后遗症。因此，我们建议测试我们的总体假设 TMEV感染后小胶质细胞产生和释放肿瘤坏死因子α的钙依赖性增加 激活神经细胞肿瘤坏死因子αR1通路，促进癫痫的产生。拟议中的实验将 使人们更好地理解病毒和免疫在急性癫痫发作中的作用 神经元和小胶质细胞功能，以及癫痫的发生。我们将使用多学科方法来测试我们的 假说，包括体内和体外最新技术，双光子显微镜，钙成像，新 转基因小鼠模型，慢性视频脑电监测和脑片电生理：1)确定 物理变化、运动性和自发钙瞬变发展的时间进程和程度 GCAMP5G在急性感染期TMEV感染小鼠小胶质细胞中的选择性表达 2)确定急性脑缺血时激活的小胶质细胞内钙瞬变的机制。 感染期和钙瞬变在细胞因子产生中的作用；以及3)决定信号是否通过 神经元型肿瘤坏死因子αR1通路参与TMEV后海马区兴奋性和惊厥活动 感染。我们预计，这些实验将为了解肿瘤坏死因子α及其受体的作用提供重要的新见解 受体、肿瘤坏死因子αR1在细胞死亡、突触传递和癫痫发生中的作用 用于预防感染所致癫痫的新型治疗干预措施的开发。

PROJECT SUMMARY: Viral infections of the central nervous system (CNS) are associated with an increased risk for seizures, status epilepticus (SE), and the development of chronic epilepsy. Our collaborative group has developed the first animal model of viral-induced epilepsy. Mice (C57BL/6) who receive intra-cerebral injections of Theiler's Murine Encephalomyelitis Virus (TMEV) display acute spontaneous seizures several days after infection, survive the initial infection and go on to develop spontaneous recurrent seizures. Furthermore, our data from the last award period using C57BL/6 mice with various cytokines or cytokine receptors knocked-out have shown that manipulations in the TNFα system can significantly alter the pathologic sequelae observed following TMEV injection. Therefore, we propose to test our overall hypothesis that following TMEV infection, calcium dependent increases in production and release of TNFα from microglia activates the neuronal TNFαR1 pathway, contributing to seizure generation. The proposed experiments will lead to a greater understanding of the role of viral and immune contributions to acute seizures, altered neuronal and microglial function, and epileptogenesis. We will use a multidisciplinary approach to test our hypothesis, including, state of the art in vivo and in vitro 2 photon microscopy, calcium imaging, novel transgenic mouse models, chronic video-EEG monitoring and brain slice electrophysiology to: 1) Determine the time course and extent of physical changes, motility, and the development of spontaneous calcium transients in microglia in TMEV infected mice during the acute infection period using GCAMP5G selectively expressed in microglia; 2) Determine the mechanisms underlying calcium transients in activated microglia during the acute infection period and the role of calcium transients in cytokine production; and 3) Determine if signaling through the neuronal TNFαR1 pathway underlies hippocampal excitability and seizure activity following TMEV infection. We anticipate that these experiments will provide important new insight into the role of TNFα and it's receptor, TNFαR1 in cell death, synaptic transmission and epileptogenesis and set the stage for the development of novel therapeutic interventions for the prevention of infection induced epilepsy.