CELLULAR AND MOLECULAR BIOLOGY CORE

获取5-羟色胺(5-HT)系统的新的细胞和分子知识是必要的 了解5-羟色胺神经生物学如何驱动临床易损性特征和成瘾。该计划的目标是 5-羟色胺和兴奋剂成瘾翻译中心的分子和细胞生物学核心(核心B) (TCSSA)是通过将临床观察与平行的细胞和分子联系起来，最大限度地提高治疗进展 生物学知识。为此，核心B将(1)建立一个稳定转基因的天然5- HT2AR和5-HT2CR以及筛选相关配体的多态和异构体(项目3) 内表型和治疗反应(项目1和2)；(2)启动和协调基因 在人类(项目1)和大鼠(项目2)中的测定，以及基因与内表型的相关性 与5-羟色胺受体和/或5-羟色胺受体的表达平行；(3)测定5-羟色胺受体的功能活性 细胞内5-HT2AR和/或5-HT2CR模型中的配体(项目3)使用细胞内(Ca)动员和/或 肌醇磷酸蓄积法和免疫沉淀法测定与G蛋白的结合 和其他结合伙伴；(4)5-HT2AR和/或5-HT2CR在特定的 大鼠的脑区由内表型(项目2)确定，使用与人类和 培养的细胞；(5)利用这些功能活性分析来确定项目3中哪些新的配体将 将在动物行为模型中进行测试(项目2)，因为我们正在塑造未来的人体实验(项目1)。 筛选以确定现有和新的配体的效力、效力和特异性(项目3)将提供 决定在啮齿动物行为(活体)模型中测试哪些新分子的客观依据(项目 2)。来自培养细胞、动物模型和人类受试者的综合数据将有助于开发一种 现有的和新的(项目3)5-HT2AR和5-HT2CR配体和意志的全面功能图 确保所有项目的进展，以开发针对可卡因的创新药物治疗方法 上瘾。 LAV摘要。目前还没有有效的、可获得的治疗兴奋剂成瘾的药物 可用。利用培养的细胞，我们将筛选新发明的药物，用于在啮齿动物身上进行测试 模拟人类吸毒并最终在临床试验中作为治疗糖尿病的新药的分析 兴奋剂成瘾。

Acquisition of new cellular and molecular knowledge of the serotonin (5-HT) system is necessary to understand how 5-HT neurobiology drives clinical vulnerability profiles and addiction. The objective of the Molecular and Cell Biology Core (Core B) of the Translational Center for Serotonin and Stimulant Addiction (TCSSA) is to maximize therapeutic advances by tying clinical observations to parallel cellular and molecular biology knowledge. To this end, Core B will (1) develop a library of stably-transfected cell lines of native 5- HT2AR and 5-HT2CR and select polymorphisms and isoforms for screening ligands (Project 3) to relate to endophenotypes and treatment responses (Projects 1 and 2); (2) initiate and coordinate genotype determinations in humans (Project 1) and rats (Project 2), and to correlate genotype with endophenotype in parallel with 5-HT^R and/or 5-HT2CR expression in platelets; (3) measure functional activity of 5-HT2R ligands (Project 3) in cellular 5-HT2AR and/or 5-HT2CR models using intracellular (Ca,++) mobilization and/or inositol phosphate accumulation assays and immunoprecipitation assays to determine association with Gproteins and other binding partners; (4) correlate 5-HT2AR and/or 5-HT2CR expression and function in specific brain regions of rats identified by endophenotype (Project 2) using assays paralleling those in humans and in cultured cells; (5) utilize these functional activity assays to determine which new ligands from Project 3 will be tested in animal behavioral models (Project 2) as we shape future experiments in humans (Project 1). Screening to determine the potency, efficacy and specificity of extant and new ligands (Project 3) will provide the objective basis for deciding which new molecules to test in the rodent behavior (in vivo) models (Project 2). Combined data from cultured cells, animal models and human subjects will aid in developing a comprehensive functional picture of extant and novel (Project 3) 5-HT2AR and 5-HT2CR ligands and will assure advancements in all Projects to develop innovative new pharmacotherapeutic approaches to cocaine addiction. Lav Abstract. No effective, accessible medication for the treatment of stimulant addiction is currently available. Using cultured cells, we will screen newly created drugs for their potential to be tested in rodent assays that model human drug-taking and ultimately in clinical trials as novel medications for treatment of stimulant addiction.