Mechanisms of astrocyte mediated glutamate homeostasis in a model of viral-induced epilepsy

病毒诱发癫痫模型中星形胶质细胞介导的谷氨酸稳态机制

基本信息

批准号：
9259393
负责人：
JAYCIE LEE LOEWEN
金额：
$ 2.76万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-01 至 2018-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9259393
关键词：
Acute Afferent Pathways Animal Model Animals Area Astrocytes Biotinylation Brain C57BL/6 Mouse Ceftriaxone Cell Death Cells Central Nervous System Infections Cicatrix Clinical Development Disease Disease model Drug Targeting Electric Stimulation Electrophysiology (science)Encephalitis Environment Epilepsy Epileptogenesis Focal Seizure Functional disorder Future Generalized seizures Glial Fibrillary Acidic Protein Gliosis Glutamate Transporter Glutamates Goals Hippocampus (Brain)Homeostasis Immunoprecipitation In Vitro Incidence Infection Inflammation Injectable Intention Lead Measures Mediating Modeling Molecular Monobactams Mus Nerve Degeneration Neuroglia Neuronal Injury Neurons Neurotransmitters Pathogenesis Pathology Patients Pharmaceutical Preparations Pharmacology Phase Play Population Process Property Regulation Reporting Resistance Role Saline Seizures Severities Slice Stratum Lucidum Surface TMEV Techniques Temporal Lobe Epilepsy Testing Tissues Toxic effect Training Viral Virus Diseases Western Blotting alternative treatment astrogliosis base beta-Lactams clinically relevant effective therapy excitotoxicity experimental study glial activation hippocampal sclerosis immunoreactivity in vivo Model mouse model multidisciplinary nervous system disorder neuron loss neurotoxicity novel novel therapeutics patch clamp prevent tool transmission process uptake

项目摘要

PROJECT SUMMARY Temporal lobe epilepsy (TLE) is the most common form of acquired epilepsy, which often results as a consequence of CNS infection and other insults. A high proportion of patients who have developed TLE following encephalitis are often resistant to current anti-seizure drugs (ASDs). As the majority of ASDs target neuronal mechanisms, the role of glial cells in epilepsy is now being actively investigated to identify alternative treatments and disease modifying therapies. The most common pathology associated with TLE, hippocampal sclerosis, involves activation and proliferation of astrocytes that form a glial scar. Astrocytes that comprise the scar have been hypothesized to play an important role in high levels of glutamate found within that tissue. The astrocyte-specific glutamate transporter GLT-1 accounts for approximately 90% of glutamate uptake in the brain. The overall goal of this project is to better understand the contribution of astrocyte-mediated glutamate homeostasis to seizures and to determine if compounds modulating the expression of GLT-1 may act as novel ASDs with potential disease modifying properties. To achieve this goal, we will conduct multidisciplinary experiments in the Theiler’s Murine Encephalomyelitis Virus (TMEV) mouse model of infection-induced TLE. This animal model, the first infection induced model of TLE, recapitulates clinical observations including neurodegeneration, formation of a glial scar, and the development of spontaneous seizures. Aim 1 will investigate changes in GLT-1 expression in reactive and glial scar astrocytes following TMEV infection. The use of electrophysiology in combination with immunoblot techniques is a superior way to observe functional alterations in glutamate regulation during the acute seizure phase in TMEV infected animals. Aim 2 will determine if a modulator of GLT-1 expression can be used as a tool to manipulate glutamate transmission and ameliorate neurotoxicity following TMEV infection. Ceftriaxone (CEF) is a safe and commonly used β-lactam antibiotic that has been shown to increase GLT-1 expression and prevent neuronal injury in other animal models of neurological disease. Thus, conducting these experiments will provide an outstanding training environment and determine the role of GLT-1 modulation in a clinically relevant epilepsy model. Additionally, the proposed experiments may provide valuable information for future disease modifying therapies for the patient with CNS infections that could lead to epilepsy.

项目摘要颞叶癫痫（TLE）是获得癫痫的最常见形式，通常会作为一个中枢神经系统感染和其他感染的结果。大部分患者患有TLE 脑炎之后通常对当前的抗塞氏菌药物（ASDS）有抵抗力。作为大多数ASD的目标神经元机制，胶质细胞在癫痫中的作用正在积极研究以鉴定替代方案治疗和疾病修饰疗法。与TLE，海马相关的最常见病理硬化症，涉及形成神经胶质疤痕的星形胶质细胞的激活和增殖。星形胶质细胞完成疤痕已被认为在该组织中发现的高水平谷氨酸盐中起重要作用。这星形胶质细胞特异性谷氨酸转运蛋白GLT-1占谷氨酸摄取的90％脑。该项目的总体目标是更好地了解星形胶质细胞介导的谷氨酸的贡献癫痫发作的稳态并确定是否调节GLT-1表达的化合物可能充当新颖具有潜在疾病修饰特性的ASD。为了实现这一目标，我们将进行多学科在Theiler的鼠脑脊髓炎病毒（TMEV）小鼠诱导的TLE的实验中。该动物模型是第一个感染的TLE模型，概括了临床观察神经变性，神经胶质疤痕的形成以及发挥癫痫发作的发展。目标1意志在TMEV感染后，研究了反应性和神经胶质疤痕星形细胞中GLT-1表达的变化。这与免疫印迹技术结合使用电生理学是观察功能的超级方法 TMEV感染动物的急性癫痫发作期间谷氨酸调节的改变。 AIM 2意志确定是否可以将GLT-1表达调节器用作操纵谷氨酸传播和 TMEV感染后的改善神经毒性。头孢曲松（CEF）是一种安全且常用的β-内酰胺已证明可以增加GLT-1表达并预防其他动物的神经元损伤的抗生素神经疾病的模型。那将进行这些实验将提供出色的培训环境并确定GLT-1调节在临床相关癫痫模型中的作用。此外，提出的实验可能会为未来的疾病修改疗法提供有价值的信息患有CNS感染的患者可能导致癫痫。