Glutamate Transporter Expression in Schizophrenia

描述(由申请者提供)：这项指导研究职业生涯奖(K08)申请的总体目标是提供创新技术方面的培训，使申请者能够在转化性心理健康研究中提出和回答最大可能的问题。这项研究培训计划有两个总体目标。第一是培训申请者使用新的尖端技术来研究死后组织中的基因表达。第二个是让申请者学习如何在啮齿类动物的大脑中进行和评估可诱导的基因缺失或过度表达。为了实现这些目标，申请者计划了一系列补充的教育和研究培训经验，包括生物统计学、生物信息学、临床流行病学、遗传学和信号转导方面的正式课程，以及与顾问一起学习电生理学、激光捕获显微镜(LCM)、定量聚合酶链式反应和病毒载体基因操作技术的扩展实验室经验。与这项申请的总体目标一致，我们设计了一个研究项目，最大限度地扩大申请人接触创新研究方法的机会，并建立在他之前对精神分裂症谷氨酸转运体转录异常的发现的基础上。这些数据表明精神分裂症患者皮质丘脑环路中谷氨酸转运体表达异常。因此，我们假设，兴奋性氨基酸转运体(EAATs)的表达和调节在精神分裂症的皮质丘脑回路中发生了改变，参与了这种疾病的精神病理学。为了评估这一假设，我们将测量EAAT mRNA在显微解剖细胞群体中的表达，并检查精神分裂症患者和对照组中EAAT蛋白的翻译后修饰。在这些人类研究的指导下，我们将表征与使用病毒介导的基因转移和/或cre-loxP重组酶系统产生的转运蛋白基因表达的皮质丘脑相关的电生理和神经化学后果。这些研究将把精神分裂症患者基因表达的变化与谷氨酸突触组成和功能的回路特异性变化联系起来。在这组实验的结论中，申请者将学习到研究严重精神疾病中基因表达和调控异常的创新的、最先进的技术，为开发独立的研究计划提供必要的工具。

DESCRIPTION (provided by applicant): The overall goal of this Mentored Research Career Award (K08) application is to provide training in innovative techniques that will permit the applicant to ask and answer the largest possible questions in translational mental health research. This research training plan has two overall objectives. The first is to train the applicant in new cutting-edge techniques to study gene expression in postmortem tissue. The second is for the applicant to learn how to perform and evaluate inducible gene deletion or overexpression in the rodent brain. To achieve these objectives, the applicant has planned a series of complementary educational and research training experiences, including formal coursework in biostatistics, bioinformatics, clinical epidemiology, genetics and signal transduction, as well as extended laboratory experiences with consultants to learn electrophysiology, laser capture microscopy (LCM), quantitative PCR, and gene manipulation techniques with viral vectors. In concert with this application's overall objectives, we have designed a research project that maximizes the applicant's exposure to innovative research methodologies and builds on his previous findings of glutamate transporter transcript abnormalities in schizophrenia. These data suggest abnormalities of glutamate transporter expression in corticothalamic circuits in schizophrenia. Accordingly, we hypothesize that expression and regulation of excitatory amino acid transporters (EAATs) is altered in corticothalamic circuits in schizophrenia, contributing to psychopathology in this illness. To evaluate this hypothesis we will measure EAAT mRNA expression in populations of microdissected cells and examine post-translational modification of EAAT proteins in subjects with schizophrenia and a control group. Guided by these human studies, we will then characterize the electrophysiological and neurochemical consequences associated with corticothalamic alterations of transporter gene expression generated using viral mediated gene transfer and/or the cre-loxP recombinase system. These studies will link identified changes in gene expression in schizophrenia with circuit specific alterations in glutamate synapse composition and function. At the conclusion of this set of experiments, the applicant will have learned innovative, state-of-the-art techniques for investigating abnormalities of gene expression and regulation in severe psychiatric illness, providing the tools needed to develop an independent research program.