NOVEL MOLECULAR SITE FOR ANTIDOPAMINERGIC EFFECTS

这些神经药理学研究的应用将研究生化 以及多巴胺受体D_1亚类的分子机制 造成他们的精神药理效应。几个假设将是 检测：D1R存在生化异质性；只有 这些受体中的一个子集与cAMP的合成有关； 这些D1多巴胺受体的一个子集与D2在功能上相互作用 受体作为同一多分子复合体的一部分；而且它是 有可能在分子水平上对这种受体类别进行建模并使用 这样的模型可以设计出对某些药物具有选择性的药物 D1R亚群。这些假设将通过以下方式进行验证 研究D1样受体表观多样性的基础，例如通过 比较[~3H]-SCH23390结合位点的出现与 多巴胺敏感型腺苷环化酶活性(DA-ACase)或 选定的药物可导致多巴胺介导的行为。减损和 药理学研究将被用来比较边缘区域和纹状体 这些功能和受体特征的基础。几个系列 刚性和半刚性多巴胺激动剂和拮抗剂的比较 采用体内和体外药理学方法。这些数据将 用于计算机辅助分子模拟研究，以模拟活性 D_1受体的位置。尽管最初这些研究将假设 单一受体，假设生物学研究最终将 提供允许至少两种类型的站点的数据(例如，一个链接到 腺苷环化酶的刺激，以及一个不是)被建模和定义。 分子模型研究将不断修改，以纳入 受体增溶、纯化和表征的数据 研究，这将是本研究的主要重点。结果就是 我们证明了SCH23390在生理上顽强地与其结合在一起 重要受体(S)，纯化实验将严重依赖 使用4-烷基苯基取代类似物的亲和层析 我们将要合成的SCH23390。净化的或可获得的 部分纯化的受体(S)会导致抗体的升高 这些蛋白质的合成，为分子生物学研究的开展奠定了基础 这些蛋白质。我们将对这些组织进行免疫组织化学定位 受体，并进行免疫中和研究。分子 生物学研究将针对所涉及的生理机制。 在D1受体的表达和调节中(例如，合成， 翻译后修饰、表达部位等)。

These neuropharmacological studies application will study the biochemical and molecular mechanisms by which the D1 subclass of dopamine receptors cause their psychopharmacological effects. Several hypotheses will be tested: that there is biochemical heterogeneity of D1 receptors; that only a subset of these receptors are linked biochemically to cAMP synthesis; that a subset of these D1 dopamine receptors interact functionally with D2 receptors as part of the same multimolecular complex; and that it is possible to model this receptor class at the molecular level and to use such models to design drugs that have selectivity for certain subpopulations of D1 receptors. These hypotheses will be tested by studying the basis for apparent multiplicity of D1-like receptors, e.g., by comparing the occurrence of [3H]-SCH23390 binding sites versus dopamine-sensitive adenylate cyclase activity (DA-ACase) or the potency of selected drugs to cause dopaminergically-mediated behaviors. Lesioning and pharmacological studies will be used to compare limbic areas to striatum on the basis of these functional and receptor characteristics. Several series of rigid and semi-rigid dopamine agonists and antagonists will be compared using both in vivo and in vitro pharmacological methods. These data will be used in computer-assisted molecular modeling studies to model the active site of D1 receptors. Although initially these studies will assume a single receptor, it is assumed that the biological studies ultimately will provide data permitting at least two types of sites (e.g., one linked to stimulation of adenylate cyclase, and one not) to be modeled and defined. The molecular modeling studies will be modified continually to incorporate the data from receptor solubilization, purification, and characterization studies which will be a major emphasis of this research. As a consequence of our demonstration that SCH23390 binds tenaciously to its physiologically important receptor(s), the purification experiments will rely heavily on affinity chromatography using a 4-alkylphenyl-substituted analog of SCH23390 that we will synthesize. The availability of purified or partially purified receptor(s) will lead to raising of antibodies against these proteins, and to the initiation of molecular biological study of these proteins. We will perform immunohistochemical localization of these receptors, and also conduct immunoneutralization studies. The molecular biological studies will be aimed at the physiological mechanisms involved in expression and regulation of D1 receptors (e.g., synthesis, posttranslational modifications, sites of expression, etc.).