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

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

展开

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.