GABA(A) RECEPTOR TRAFFICKING AND EPILEPTOGENESIS

描述（由申请人提供）：此提案描述了一个为期5年的培训计划，以发展神经科学领域的搜索职业。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而富有成效的博士后培训，现在他正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。本课题将探讨GABAA受体（GABAAR）在癫痫发生过程中组装和转运的分子调控机制。先前的研究表明，在颞叶癫痫（TLE）动物模型中，GABAAR亚基表达、突触定位和功能损伤发生了改变，但引发这些变化的分子机制尚不清楚。拟议的研究旨在阐明在癫痫发生过程中，GABAAR与运输、锚定和/或聚类所需的支架蛋白的相互作用是否被破坏。由于BDNF长期以来一直与癫痫发生机制有关，最近发现其调节GABAAR的运输和表达，因此还将进行实验来研究BDNF在调节蛋白质-蛋白质相互作用、运输和GABAAR锚定中的作用。最后，将从对照组和癫痫大鼠中分离天然GABAAR多蛋白复合物，并使用蛋白质组学方法鉴定其成分以生成GABAAR相关蛋白质组。一旦完成，这将是第一个详细分析在癫痫发生过程中通过蛋白质-蛋白质相互作用调节GABAAR功能的研究。这些研究代表了阐明癫痫中GABAAR失调的分子成分的基本步骤，并可能为癫痫治疗提供新的靶点和新方法。艾米·布鲁克斯-卡亚尔博士将担任首席研究员科学发展的主要导师。Brooks-Kayal博士是癫痫病发生过程中GABAAR作用研究的公认领导者。布鲁克斯-卡亚尔博士是科罗拉多大学儿科教授，也是儿童医院神经内科主任。她培养了许多研究生、博士后和初级教师。Mark Dell’acqua博士将担任该奖项的联合导师。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，并将提供开发贩运和成像研究的专业知识。科罗拉多大学，特别是加州大学丹佛分校的转化癫痫研究项目，是一个理想的指导独立科学事业的环境，结合了一个合作的环境，在神经科学项目和药理学，生理学，儿科和药物科学系拥有广泛的教师，这将最大限度地发挥主要研究者发展学术事业的潜力。

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. PUBLIC HEALTH RELEVANCE: Approximately 200,000 new cases of seizures and epilepsy occur ery ear and an estimated $15.5 billion dollars are needed to provide care for the ~3 million Americans that suffer chronic seizures or epilepsy. The most common form of epilepsy, Temporal Lobe Epilepsy, is in many cases refractory to current medications. The proposed studies aim to develop a better understanding of the molecular mechanisms that trigger epilepsy development to provide new therapeutic targets and generate novel strategies to ameliorate epilepsy progression.