Dysregulation of Glutamine Synthetase in Human Temporal Lobe Epilepsy

人颞叶癫痫中谷氨酰胺合成酶的失调

基本信息

批准号：
7561630
负责人：
TORE EID
金额：
$ 17.33万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-01 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7561630
关键词：
Acute Address Animal Model Animals Antiepileptic Agents Area Arts Astrocytes Basic Science Binding Biological Assay Brain Brain Injuries Cell Culture Techniques Chronic Cultured Cells Development Disease Down-Regulation Drug resistance Enzymes Epigenetic Process Epilepsy Etiology Flow Cytometry Gene Expression Genes Genetic Techniques Glutamate-Ammonia Ligase Glutamates Glutamine Goals Hippocampus (Brain)Histone Deacetylation Human Immunologic Techniques In Situ Hybridization In Vitro Intervention Knowledge Learning Ligase Mass Spectrum Analysis Mentors Methodology Modeling Molecular Biology Molecular Genetics Patients Physicians Poison Post-Translational Protein Processing Proteins Proteomics Rattus Recurrence Regulation Research Personnel Research Training Resected Scientist Seizures Series Specimen System Techniques Temporal Lobe Epilepsy Testing Time Transcriptional Regulation Translational Research Two-Dimensional Gel Electrophoresis abstracting chromatin immunoprecipitation experience laser capture microdissection mRNA Expression novel prevent research study transcription factor

项目摘要

DESCRIPTION (provided by applicant): Dysregulation of Glutamine Synthetase in Human Temporal Lobe Epilepsy: The main objective of this K08 proposal is to facilitate the applicant's development towards independence as a physician-scientist investigator in basic science/translational research. This will be achieved through a series of complementary educational and mentored research training experiences in which the applicant will learn state-of-the-art methodologies in molecular biology, proteomics, cell culture and flow cytometry. The scientific question addressed by these methodologies is: what is the mechanism underlying the down-regulation of glutamine synthetase in the hippocampus of patients with mesial temporal lobe epilepsy? This question is important because experimental inhibition of glutamine synthetase in the hippocampus of animals causes epileptic seizures followed by cessation of the seizures once the enzyme has been restored. The specific aims of this proposal are: 1) Analysis of transcriptional regulation of glutamine synthetase using laser capture microdissection, quantitative real time PCR, and in situ hybridization on surgically resected human epilepsy hippocampi. 2) Analysis of posttranslational modification of glutamine synthetase by 2D gel electrophoresis, immunological techniques, and mass spectrometry. 3) Establish an in vitro system for experimental studies of glutamine synthetase regulation, using acute and chronic cultures of astrocytes from human epileptogenic hippocampi combined with flow cytometric analysis of the cultured cells. Layman's Abstract: People with temporal lobe epilepsy have low levels of brain glutamine synthetase, an enzyme that transforms glutamate to the non toxic chemical glutamine. The goal is to understand why the enzyme is down-regulated, so that new antiepileptic treatments targeting glutamine synthetase can be developed.

描述（由申请人提供）：人类颞叶癫痫中谷氨酰胺合成酶的失调：该K08提案的主要目的是促进申请人发展成为基础科学/翻译研究的医师科学家研究者独立的独立性。这将通过一系列互补的教育和指导的研究培训经验来实现，其中申请人将学习分子生物学，蛋白质组学，细胞培养和流式细胞术中的最新方法。这些方法论提出的科学问题是：介体颞叶癫痫患者的海马中谷氨酰胺合成酶下调的基础机制是什么？这个问题很重要，因为在动物海马中对谷氨酰胺合成酶的实验抑制会引起癫痫发作，然后恢复酶后停止癫痫发作。该提案的具体目的是：1）使用激光捕获微分解，定量实时PCR以及对手术切除的人类癫痫海马乳房的原位杂交分析谷氨酰胺合成酶的转录调节。 2）通过2D凝胶电泳，免疫学技术和质谱法分析谷氨酰胺合成酶的翻译后修饰。 3）建立一种体外系统，用于使用人类癫痫源性海马的星形胶质细胞的急性和慢性培养谷氨酰胺合成酶调节的实验研究，并结合了培养细胞的流式细胞仪分析。外行的摘要：颞叶癫痫患者的脑谷氨酰胺合成酶含量较低，这种酶是将谷氨酸转化为非毒性化学谷氨酰胺的酶。目的是了解为什么酶被下调，以便可以开发出靶向谷氨酰胺合成酶的新抗癫痫疗法。