GABA(A) RECEPTOR TRAFFICKING AND EPILEPTOGENESIS

GABA(A) 受体贩运和癫痫发生

• 批准号: 8440282
• 负责人: Marco I. Gonzalez
• 金额: $ 15.81万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8440282
• 关键词: Acute; Adult; American; Animal Model; Antibodies; Award; Brain; Brain-Derived Neurotrophic Factor; Caring; Cell membrane; Cell surface; Chronic; Co-Immunoprecipitations; Colorado; Complex; Computer Simulation; Coupled; Detergents; Development; Dominant-Negative Mutation; Ear; Ensure; Environment; Epilepsy; Epileptogenesis; Faculty; Functional disorder; GABA-A Receptor; Goals; Hippocampus (Brain); Human; Image; Immunofluorescence Immunologic; Immunoprecipitation; Impairment; Individual; Knowledge; Label; Long-Term Effects; Mass Spectrum Analysis; Mediating; Mentors; Mentorship; Molecular; Molecular and Cellular Biology; Multiprotein Complexes; Neurology; Neurons; Neurosciences; Pathway Analysis; Pediatric Hospitals; Pediatrics; Pennsylvania; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Philadelphia; Physiological; Physiology; Postdoctoral Fellow; Principal Investigator; Property; Proteins; Proteome; Proteomics; RNA Interference; Rat Protein; Rattus; Recurrence; Refractory; Regulation; Research; Research Personnel; Role; Scaffolding Protein; Science; Seizures; Signal Transduction; Slice; Specificity; Status Epilepticus; Synapses; Synaptic plasticity; Temporal Lobe Epilepsy; Testing; Time; Training; Training Programs; Transmembrane Domain; Trypsin; Universities; Western Blotting; career; functional disability; genetic regulatory protein; graduate student; human RIPK1 protein; human disease; mRNA Expression; new therapeutic target; novel; novel strategies; post-doctoral training; professor; programs; protein complex; protein expression; protein protein interaction; public health relevance; receptor; receptor function; research study; skills; trafficking

DESCRIPTION (provided by applicant): This proposal describes a 5-year training program to develop search career in Neuroscience. The Principal Investigator has conducted intensive and productive postdoctoral training at the University of Pennsylvania and The Children's Hospital of Philadelphia, and now is looking to expand his scientific skills by integrating his previous knowledge in cellular and molecular biology with animal models of human disease. The current proposal will explore the molecular mechanisms regulating GABAA Receptors (GABAAR) assembly and trafficking during epileptogenesis. Previous studies have demonstrated altered GABAAR subunit expression, synaptic localization and functional impairments in animal models of Temporal Lobe Epilepsy (TLE), but the molecular mechanisms triggering these changes are unknown. The proposed studies are aimed at elucidating if during epileptogenesis the interaction of GABAAR with scaffolding proteins necessary for trafficking, anchoring, and/or clustering are disrupted. Since BDNF has long been implicated in mechanisms of epileptogenesis and recently found to regulate GABAAR trafficking and expression, experiments will also be conducted to examine the role of BDNF on the regulation of protein- protein interactions, trafficking, and anchoring of GABAAR. Finally, native GABAAR multiprotein complexes will be isolated from control and epileptic rats and their components identified using a proteomic approach to generate GABAAR-associated proteomes. Once completed, this will be the first detailed analysis dealing with the regulation of GABAAR function by protein-protein interactions during epileptogenesis. These studies represent a fundamental step to elucidate the molecular components responsible for GABAAR dysregulation in epilepsy and are likely to provide new targets and novel approaches for epilepsy treatment. Dr. Amy Brooks-Kayal will act as the Primary Mentor for the scientific development of the Principal Investigator. Dr. Brooks-Kayal is a recognized leader in the study of the role of GABAAR during epileptogenesis. Dr. Brooks-Kayal is a Professor of Pediatrics at the University of Colorado and Chief of the Division of Neurology at the Children's Hospital. She has trained numerous graduate students, Postdoctoral fellows and junior faculty. Dr. Mark Dell'Acqua will be the Co-Mentor for this award. Dr. Dell'Acqua has expertise in the study of the organization of signaling complexes by anchoring proteins and synaptic plasticity, and will provide expertise to develop the trafficking and imaging studies. The University of Colorado, and in particular the Translational Epilepsy Research Program at UC Denver are an ideal setting for mentorship towards an independent scientific career, incorporating a collaborative environment that has broad faculty in the Neuroscience Program and the Departments of Pharmacology, Physiology, Pediatrics and Pharmaceutical Sciences that will maximize the potential of the Primary Investigator to develop an academic career.

描述（由申请人提供）：本提案描述了一个为期5年的培训计划，以发展神经科学的搜索职业生涯。首席研究员在宾夕法尼亚大学和费城儿童医院进行了密集和富有成效的博士后培训，现在正在寻求通过将他以前在细胞和分子生物学方面的知识与人类疾病的动物模型相结合来扩展他的科学技能。 目前的建议将探讨癫痫发生过程中调控GABAA受体（GABAAR）组装和运输的分子机制。以往的研究表明，颞叶癫痫（TLE）的动物模型中GABAAR亚基的表达，突触定位和功能障碍的改变，但触发这些变化的分子机制是未知的。拟议的研究的目的是阐明，如果在癫痫发作的GABAAR与骨架蛋白的相互作用所需的运输，锚定，和/或集群被破坏。由于BDNF长期以来一直与癫痫发生机制有关，最近发现其可调节GABAAR的运输和表达，因此还将进行实验以检查BDNF对GABAAR的蛋白质-蛋白质相互作用、运输和锚定的调节作用。最后，天然GABAAR多蛋白复合物将从对照组和癫痫大鼠中分离，并使用蛋白质组学方法鉴定其组分以产生GABAAR相关蛋白质组。一旦完成，这将是第一次详细分析癫痫发生过程中蛋白质-蛋白质相互作用对GABAAR功能的调节。这些研究代表了阐明癫痫中GABAAR失调的分子组分的基本步骤，并可能为癫痫治疗提供新的靶点和新的方法。 Amy Brooks-Kayal博士将担任主要研究者科学发展的主要导师。Brooks-Kayal博士是研究GABAAR在癫痫发生过程中的作用的公认领导者。Brooks-Kayal博士是科罗拉多大学的儿科教授和儿童医院神经科主任。她培养了许多研究生，博士后研究员和初级教师。Mark Dell'Acqua博士将担任该奖项的共同导师。Dell'Acqua博士在通过锚定蛋白和突触可塑性研究信号复合物的组织方面具有专业知识，并将为开发贩运和成像研究提供专业知识。科罗拉多大学，特别是转化癫痫研究计划在加州大学丹佛分校是一个理想的设置导师走向独立的科学事业，结合了一个协作环境，具有广泛的教师在神经科学计划和药理学，生理学，儿科学和药物科学的部门，将最大限度地发挥主要研究者的潜力，发展学术生涯。