CORTICAL CIRCUITRY RELATED TO NEUROTRANMISSION PROTEINS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。这项工作将前额叶皮质回路与多巴胺神经传递的分子成分联系起来。这一结果将与精神分裂症患者的工作记忆回路有关。工作记忆功能依赖于d1受体激活的最佳水平，并受到d1受体过度和过度刺激的影响。希望我们的结果将有助于更好地理解D1受体对工作记忆的复杂调节机制，并最终更好地理解抗精神病药物的作用。我们将确定终止于含有D1和蛋白磷酸酶-1(PP1)亚型PP1(α)和PP1(Gamma)1的脊椎的传入来源。我们将神经解剖束示踪剂注射到年轻成年猕猴的大脑中，以标记前额叶皮质的顶叶、丘脑和后扣带回传入，并在前额叶皮质内标记局部水平轴突。我们正在使用适合于电子显微镜的双重标记技术来染色标记的轴突以及D1、PP1(α)或PP1(Gamma)1。我们正在使用连续切片电子显微镜来确定标记的轴突的突触后靶是否包含这些蛋白。这些蛋白质的分布细分了皮质脊椎，结果将提示皮层回路的某些方面对工作记忆功能至关重要，并扩大了我们对皮质回路专门化机制的理解。我们使用包埋后免疫金标记法检测了PP1靶向蛋白(刺亲素和神经肽)以及PP1(α)和PP1(Gamma1)在脊椎中的分布。我们的数据表明，D1R在不同的神经元区段中可以接触到不同的信号转导蛋白的补体，并提示两种PP1亚型的功能差异。最后，我们使用单标记法和包埋前双标记法研究了I组mGluR(mGluR1a和mGluR5)的亚细胞定位，并确定了它们在彼此之间、在猴皮质和大鼠皮质之间以及在皮质和基底节之间的分布的一些差异。该项目的资金于2005年8月结束，目前处于无费用延期年。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This work relates prefrontal cortical circuitry with the molecular components of dopamine neurotransmission. The results will be related to the circuitry of working memory, which is dysfunctional in schizophrenia. Working memory function depends on an optimal level of D1 receptor activation and is impaired by both over and under stimulation of D1 receptor. It is hoped that our results will lead to a better understanding of the mechanism of the D1 receptor's complex modulation of working memory, and ultimately to a better understanding of antipsychotic drug action. We will identify the source of afferents that terminate onto spines that contain D1 and protein phosphatase-1 (PP1) isoforms PP1(alpha) and PP1(gamma)1. We have injected neuroanatomical tract tracers into the brains of young adult macaque monkeys to label parietal, thalamic and posterior cingulate afferents to prefrontal cortex, and within prefrontal cortex to label local horizontal axons. We are using double-labeling techniques appropriate for electron microscopy to stain the labeled axons and either D1, PP1(alpha) or PP1(gamma)1. We are using serial section electron microscopy to determine if the postsynaptic targets of labeled axons contain these proteins. The distribution of these proteins subdivides cortical spines, and the results will suggest aspects of cortical circuitry that are critical for working memory function and extend our understanding of the mechanisms by which cortical circuitry may be specialized. We have used post-embedding immunogold labeling to examine the distribution of PP1 targeting proteins (spinophilin and neurabin) and PP1(alpha) and PP1(gamma1) in spines. Our date indicates that the D1 receptor has access to different complements of signal transduction proteins in different neuronal compartments and suggests functional differences for the two PP1 isoforms. Finally we have used single and pre-embedding double label methods to examine the subcellular localization of group I mGluRs (mGluR1a and mGluR5) and determined a number of differences in their distribution, both between each other, between monkey cortex and rat cortex and between cortex and basal ganglia. The funding for this project ended in August of 2005 and it is currently in a no cost extension year.