Glutamate Transporter Expression in Schizophrenia

描述（由申请人提供）：这个指导研究职业奖（K08）申请的总体目标是提供创新技术的培训，使申请人能够在转化心理健康研究中提出和回答最大可能的问题。本研究培训计划有两个总体目标。第一个是培养申请人在研究死后组织中基因表达的新前沿技术。第二是让申请人学习如何在啮齿动物大脑中执行和评估诱导基因缺失或过度表达。为了达到这些目标，申请人计划了一系列补充的教育和研究培训经历，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PCR和病毒载体基因操作技术的扩展实验室经验。与此申请的总体目标相一致，我们设计了一个研究项目，最大限度地提高申请人对创新研究方法的接触，并建立在他之前对精神分裂症中谷氨酸转运蛋白转录异常的发现的基础上。这些数据提示精神分裂症患者皮质丘脑回路中谷氨酸转运蛋白表达异常。因此，我们假设精神分裂症患者皮质丘脑回路中兴奋性氨基酸转运蛋白（EAATs）的表达和调控发生改变，从而导致精神病理。为了验证这一假设，我们将测量微解剖细胞群体中EAAT mRNA的表达，并检测精神分裂症患者和对照组中EAAT蛋白的翻译后修饰。在这些人体研究的指导下，我们将描述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改变相关的电生理和神经化学后果。这些研究将把精神分裂症中基因表达的变化与谷氨酸突触组成和功能的电路特异性改变联系起来。在这组实验结束时，申请人将学习到用于研究严重精神疾病中基因表达异常和调控的创新的、最先进的技术，提供开发独立研究计划所需的工具。

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.