Hallucinogens and 5-HT Receptors: Mechanisms and Effects

描述（由申请人提供）：致幻剂作用呈现复杂的情况，因为它们（a）对特定靶点（如5-HT 2受体亚型（5-HT 2A、5-HT 2C））引起与其他激动剂不同的反应;（B）调节特定神经元中的多种信号传导途径;（c）在其行为效应中涉及几个脑区。在这个项目的前一阶段，我们已经开发了一种多尺度方法来解决这个复杂的问题，将分子和细胞信号事件的见解与参与行为生成的神经元和大脑区域中的更大尺度过程联系起来。我们还开发了强大的遗传动物模型和蛋白质工程构建体，可用于在体外和体内探测这种专门设计的构建体的后果。本PPG更新申请的研究计划基于三个项目中迭代研究的跨学科方案，提出了此类多尺度研究。协调和综合的研究跨越规模和组织：从野生型和突变型GPCR的活化形式的分子结构及其在信号通路中的相互作用，这是在项目1中研究;到基于定量基因组学的称为转录组指纹的方法，该方法反映了通过这些受体的差异信号应答，以及在细胞信号传导水平上识别参与致幻剂特异性反应的神经元和特异性分子机制，这在项目2中进行;并在项目3中进一步研究具有脑区域特异性操纵5-HT 2AR及其与其他受体和信号介质相互作用的转基因小鼠。在项目1中从生物信息学、基于结构的分析和模拟中设计的受体及其信号相关蛋白的新构建体将在项目2中表达，以（i）研究致幻剂信号特异性的细胞机制;（ii）测试关于受体二聚化和与项目1中产生的支架蛋白相互作用的机制假设;和（iii）选择在机制上最具揭示性的那些用于项目#3中的敲入/敲除和区域特异性恢复和缺失实验，以阐明与特定机制的支持或破坏相关的行为表型。通过其综合性的结构和重点，这项研究计划预计将继续对理解药物滥用的一般意义，以及对药物成瘾的新的基于GPCR的治疗方法的设计。 程序特征

DESCRIPTION (provided by applicant): Hallucinogen actions present a complicated picture because they (a) cause responses that differ from those of other agonists on specific targets such as 5-HT2 receptor subtypes (5-HT2A, 5- HT2C); (b) modulate multiple signaling pathways in specific neurons; and (c) involve several brain regions in their behavioral effects. In the previous period of this Program Project we have developed a multi-scale approach to this complex problem that connects insights about molecular and cellular signaling events to larger scale processes in the neurons and brain regions involved in the generation of the behaviors. We have also developed the powerful genetic animal models and the protein engineering constructs that can be used to probe the consequences of such specifically designed constructs in vitro and in vivo. The research plan of this PPG renewal application presents such a multi-scale investigation, based on an interdisciplinary protocol of iterative studies in the three Projects. The coordinated and integrated studies span scales of size and organization: from molecular structures of activated forms of the wild type and mutant GPCRs and their interactions in the signaling pathways, that are studied in Project 1; to the quantitative genomics-based approach termed Transcriptome Fingerprinting that reflects differential signaling responses through these receptors, and the identification of neurons and specific molecular mechanisms at the cell signaling level that are involved in hallucinogen-specific responses, pursued in Project 2; and onward to genetically modified mice with brain-region-specific manipulations of the 5-HT2AR and its interactions with other receptors and signaling mediators, in Project 3. New constructs of the receptors and their signaling-associated proteins, engineered in Project 1 from bioinformatics, structure-based analysis and simulations, will be expressed in Project 2 to (i) study of the cellular mechanisms underlying hallucinogen signaling specificity; (ii) test mechanistic hypotheses about receptor dimerization and interactions with scaffolding proteins generated in Project 1; and (iii) select the mechanistically most revealing ones for knock-in/knock-out and region-specific restoration and deletion experiments in Project #3, in order to elucidate the behavioral phenotypes associated with support or disruption of specific mechanisms. Through its integrative construct and focus, this research plan is expected to continue to have significant implications for understanding drug abuse in general, as well as on the design of new GPCR-based therapeutics for drug addiction. PROGRAM CHARACTERISTICS