Hallucinogens and 5-HT Receptors: Mechanisms and Effects

描述(申请人提供)：迷幻剂的作用呈现出一幅复杂的图景，因为它们(A)引起与其他激动剂不同的对特定靶点的反应，例如5-HT2受体亚型(5-HT2A，5-HT2C)；(B)调节特定神经元中的多个信号通路；以及(C)在其行为效应中涉及几个大脑区域。在本计划项目的前一阶段，我们开发了一种多尺度方法来解决这个复杂的问题，将对分子和细胞信号事件的见解与参与行为产生的神经元和大脑区域的更大规模的过程联系起来。我们还开发了强大的基因动物模型和蛋白质工程结构，可以用来探索这种特殊设计的结构在体外和体内的后果。这个PPG更新应用的研究计划提出了这样一个多尺度的调查，基于三个项目中迭代研究的跨学科协议。协调和综合的研究跨越规模和组织范围：从项目1中研究的野生型和突变型GPCRs的激活形式的分子结构及其在信号通路中的相互作用；到项目2中追求的基于定量基因组学的方法，称为转录组指纹图谱，它通过这些受体反映不同的信号反应，以及在细胞信号水平上识别参与幻觉剂特异性反应的神经元和特定的分子机制；在项目3中，继续到对5-HT2AR及其与其他受体和信号介体的相互作用进行脑区特异性操作的转基因小鼠。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结构将在项目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以及与项目1中产生的支架蛋白相互作用的机械假说；以及(Iii)为项目3中的敲入/敲除以及特定区域的恢复和删除实验选择机制上最具启发性的基因，以阐明与支持或破坏特定机制相关的行为表型。通过其综合结构和重点，这项研究计划预计将继续对总体上了解药物滥用以及设计基于GPCR新的药物成瘾疗法产生重大影响。 节目特点

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