GABA(A) RECEPTOR TRAFFICKING AND EPILEPTOGENESIS

描述(由申请人提供)：本建议书描述了一项为期5年的培训计划，旨在发展神经科学领域的搜索职业生涯。这位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。目前的提案将探索癫痫发生过程中GABAA受体(GABAAR)组装和运输的分子机制。以往的研究表明，在颞叶癫痫(TLE)的动物模型中，GABAAR亚单位的表达、突触定位和功能损伤都发生了变化，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/或聚集所必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的发生机制有关，并且最近被发现调节GABAAR的运输和表达，因此，我们还将通过实验来研究BDNF在调节GABAAR的蛋白质-蛋白质相互作用、运输和锚定方面的作用。最后，将从对照组和癫痫大鼠中分离出天然的GABAAR多蛋白复合体，并使用蛋白质组学方法鉴定其成分，以产生GABAAR相关蛋白质组。一旦完成，这将是第一个关于癫痫发生过程中通过蛋白质-蛋白质相互作用调节GABAAR功能的详细分析。这些研究代表了阐明癫痫中GABAAR失调的分子组成的基本步骤，并可能为癫痫的治疗提供新的靶点和新的方法。艾米·布鲁克斯-卡亚尔博士将担任首席调查员科学发展的主要导师。布鲁克斯-卡亚尔博士在研究GABAAR在癫痫发生中的作用方面是公认的领导者。布鲁克斯-卡亚尔博士是科罗拉多大学儿科教授，也是儿童医院神经科主任。她培养了大量的研究生、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质和突触可塑性来研究信号复合体的组织方面拥有专业知识，并将提供专业知识来开发运输和成像研究。科罗拉多大学，尤其是加州大学丹佛分校的转化性癫痫研究项目，是指导独立科学生涯的理想环境，整合了神经科学项目和药理学、生理学、儿科和制药科学系的广泛教员的合作环境，将最大限度地发挥首席研究员发展学术生涯的潜力。

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.