MOLECULAR REGULATION OF GABAA RECEPTORS IN THE AMYGDALA

该子项目是利用资源的许多研究子项目之一 由NIH/NCRR资助的中心拨款提供。次级项目的主要支助 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 表示子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 我们在过去一年中取得了相当大的进展。 我们已经完成了所有与恐惧相关的方面，并获得了关于BZD治疗的急性、慢性和停药的试点数据。 我们已经检查了不同水平的试验药物对运动（镇静），PTZ诱导的癫痫发作（抗惊厥作用），以及在单独的动物队列中获得巴甫洛夫背景恐惧（健忘症）的影响。 我们还确定了PTZ剂量水平（25-100 mg/kg）产生癫痫发作的可靠性和允许检测实验诱导的变化的水平。 我们一直致力于最大限度地提高我们在体外和体内用慢病毒载体沉默Gephyrin蛋白的能力。 我们继续使用基因敲除小鼠品系，并致力于用慢病毒介导的GAD 65表达“拯救”GAD 65。 我们已经完成了慢病毒载体的工作，令人信服地减少GAD 67的表达，并已开始使用它也用于行为研究。 我们最近发表了（Heldt et al.，2010，J Neurosci）数据，证明了GABA（A）α 1缺失的行为效应，特别是从杏仁核。 我们还率先使用原代杏仁核培养物来检查GABA（A）受体调节和再循环（Mou et al.，2010，Neuroscience）。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We have made considerable progress in the past year. We have completed all of the fear-related aspects and have obtained pilot data on the acute, chronic, and withdrawal of BZD treatment. We have examined the effects of the various levels of test drugs on motor locomotion (sedation), PTz-induced seizures (anticonvulsant effects), and the acquisition of Pavlovian contextual fear (amnesia) in separate cohorts of animals. We also determined what PTZ dose level (25-100 mg/kg) produced seizures both reliably and at levels which allow detection of experimentally-induced changes. We have been working to maximize our ability to silence the Gephyrin protein in vitro and in vivo with lentiviral vectors. We have continued to work with the knockout mouse line, and have worked to "rescue" GAD65 with lentivirus-mediated GAD65 expression. We have completed work on a lentiviral vector that convincingly decreases GAD67 expression, and have begun using this too for behavioral studies. We have recently published (Heldt et al., 2010, J Neurosci) data demonstrating behavioral effects of GABA(A)alpha1 deletions specifically from amgydala. We have also pioneered the use of primary amgydala cultures to examine GABA(A) receptor modulation and recycling (Mou et al., 2010, Neuroscience).