Role of microglia in a novel model of temporal lobe epilepsy

小胶质细胞在颞叶癫痫新模型中的作用

• 批准号: 10392925
• 负责人: Karen S Wilcox
• 金额: $ 33.36万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392925
• 关键词: AMPA Receptors; Acute; Address; Animal Model; Animals; Astrocytes; Award; Awareness; Behavioral; Biological Assay; Biotinylation; Brain; C57BL/6 Mouse; Calcium; Cell Death; Central Nervous System Infections; Central Nervous System Viral Diseases; Child; Chronic; Clinical; Communities; Convulsants; Cytokine Receptors; Data; Development; Electroencephalography; Electrophysiology (science); Environment; Enzyme-Linked Immunosorbent Assay; Epilepsy; Epileptogenesis; Exhibits; Febrile Convulsions; Generations; HIV; Hippocampus (Brain); Human; Image; Immune; In Vitro; Incidence; Infection; Infection prevention; Inflammatory; Injections; Japanese encephalitis virus; Knock-out; Knockout Mice; Life; Limbic System; Link; LoxP-flanked allele; Membrane; Messenger RNA; Microglia; Modeling; Monitor; Morphology; Mus; Necrosis; Nerve Degeneration; Neuroglia; Neurons; Pathologic; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Prevention; Process; Production; Prosencephalon; Proteins; Reactive Oxygen Species; Recurrence; Research; Resected; Resolution; Risk; Role; Seizures; Signal Transduction; Simplexvirus; Slice; Source; Status Epilepticus; Synaptic Transmission; System; TMEV; TNF gene; TNFRSF1A gene; TNFRSF1B gene; Temporal Lobe Epilepsy; Testing; Therapeutic; Time; Transgenic Mice; Translational Research; Uncertainty; Universities; Utah; Viral; Virus Diseases; West Nile virus; Work; acute infection; cell motility; comorbidity; conditional knockout; cytokine; experimental study; human tissue; in vivo; ineffective therapies; innovation; insight; interdisciplinary approach; mRNA Expression; mouse model; neuron loss; novel; novel therapeutic intervention; novel therapeutics; prevent; protein expression; receptor; receptor expression; selective expression; trafficking; tumor; two photon microscopy; two-photon

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.

项目摘要：中枢神经系统 (CNS) 的病毒感染与病毒感染增加有关 癫痫发作、癫痫持续状态 (SE) 和发展为慢性癫痫的风险。我们的协作组有 开发了第一个病毒引起的癫痫动物模型。接受脑内注射的小鼠 (C57BL/6) 注射泰勒氏鼠脑脊髓炎病毒 (TMEV) 会导致急性自发性癫痫发作 感染后几天，从最初的感染中幸存下来，并继续发展为自发性复发性癫痫发作。 此外，我们使用具有各种细胞因子或细胞因子的 C57BL/6 小鼠在上一个奖励期获得的数据 受体敲除表明，TNFα系统的操作可以显着改变病理学 TMEV 注射后观察到的后遗症。因此，我们建议检验我们的总体假设 TMEV 感染后，小胶质细胞中 TNFα 的产生和释放呈钙依赖性增加 激活神经元 TNFαR1 通路，导致癫痫发作。拟议的实验将 导致人们更好地了解病毒和免疫对急性癫痫发作的作用，改变 神经元和小胶质细胞功能以及癫痫发生。我们将使用多学科方法来测试我们的 假设，包括体内和体外最先进的 2 光子显微镜、钙成像、新颖 转基因小鼠模型、慢性视频脑电图监测和脑切片电生理学：1) 确定 身体变化、运动和自发钙瞬变发展的时间过程和程度 使用 GCAMP5G 在急性感染期间选择性表达的 TMEV 感染小鼠的小胶质细胞中 小胶质细胞； 2）确定急性期激活小胶质细胞钙瞬变的机制 感染期和钙瞬变在细胞因子产生中的作用； 3) 确定信号是否通过 神经元 TNFαR1 通路是 TMEV 后海马兴奋性和癫痫发作活动的基础 感染。我们预计这些实验将为 TNFα 的作用及其作用提供重要的新见解。 受体、TNFαR1 在细胞死亡、突触传递和癫痫发生中的作用，并为 开发新的治疗干预措施来预防感染诱发的癫痫。