CELLULAR AND MOLECULAR BIOLOGY CORE

获取血清素（5-HT）系统的新细胞和分子知识是必要的， 了解5-HT神经生物学如何驱动临床脆弱性和成瘾。的目的 5-羟色胺和兴奋剂成瘾转化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生物学相结合， 生物学知识。为此，核心B将（1）开发稳定转染的天然5-氨基-1，2-二硫代半乳糖苷酶细胞系的文库。 HT 2AR和5-HT 2CR，并选择多态性和同种型用于筛选配体（项目3）， 内表型和治疗反应（项目1和2）;（2）启动和协调基因型 在人类（项目1）和大鼠（项目2）中进行基因型测定，并将基因型与内表型相关联， 与血小板中5-HT 2 R和/或5-HT 2CR表达平行;（3）测量5-HT 2 R和/或5-HT 2CR的功能活性， 在细胞5-HT 2AR和/或5-HT 2CR模型中使用细胞内（Ca，++）动员和/或 肌醇磷酸积累试验和免疫沉淀试验，以确定与G蛋白的结合 （4）将5-HT 2AR和/或5-HT 2CR的表达和功能与特异性结合蛋白的表达和功能相关联。 通过内表型（项目2）确定的大鼠脑区，使用与人类和 （5）利用这些功能活性测定来确定来自项目3的哪些新配体将 在动物行为模型中进行测试（项目2），因为我们塑造了未来的人类实验（项目1）。 筛选以确定现有和新配体的效力、功效和特异性（项目3）将提供 决定在啮齿动物行为（体内）模型中测试哪些新分子的客观基础（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-HT 2AR和5-HT 2CR配体的全面功能图片，并将 确保所有项目取得进展，以开发可卡因的创新性新制药方法 成瘾 拉夫摘要。目前还没有有效的、可获得的治疗兴奋剂成瘾的药物。 available.利用培养的细胞，我们将筛选新开发的药物，以在啮齿动物中进行测试 模拟人类药物服用并最终在临床试验中作为治疗 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.