Component for Institution # 264313

对药物滥用和成瘾的研究需要采取多学科的方法。生化 药理学核心为不从事生化药理学的研究人员提供服务 测定。本核心中提供的技术允许询问与药物相关的受体， 虐待和他们的信号。大多数使用核心的研究人员要么是行为科学家， 化学家核心小组提供的服务对这些调查人员很有价值， 研究并发表了许多出版物。核心还提供了赠款的初步数据 申请和几个NIH赠款被授予。内核的滥用药物受体 迄今为止研究的包括阿片受体（MOPR、DOPR、KOPR、NOPR）、大麻素（CB 1和 CB 2）受体、烟碱胆碱能受体（nAChR）以及多巴胺和σ受体。在 在所要求的资金期限内，核心将执行以下服务：（1）放射性配体结合到 （2）对G受体的评估 蛋白偶联受体（GPCR）激活，包括cAMP水平，[35 S]GTPγS结合，p44/42 MAP 激酶磷酸化，β-抑制蛋白募集;（3）使用GPCR的激动剂偏倚的测定 [35 S]GTPγS结合或cAMP和β-抑制蛋白募集和CRISPR细胞系;（4） 放射性配体与阿片和多巴胺受体的结合和（5）阿片激动剂的放射自显影- 促进脑切片中[35 S]GTPγS结合;（6）MOPR、KOPR和 （7）MOPR、DOPR和DOPR的内化; 培养细胞中的KOPR和CB 1以及小鼠脑中的KOPR; MOPR、KOPR和phospho-KOPR;（9）培训其他实验室的人员。为 在申请资助期间，核心将与NIH资助的至少15个合作项目 全国各地的调查人员。创新之处在于拟议的研究，这些研究将解决广泛的 与药物滥用有关的各种问题，包括说明对青少年和成人的影响 慢性暴露于吗啡、可卡因或尼古丁对MOPR、多巴胺和/或nAChR表达的影响 和/或信号传导;寻找KOPR激动剂， KOPR磷酸化（可能偏向G蛋白）; AMP活化蛋白激酶（AMPK）的作用 MOPR和nAChR表达的缺失;筛选CB 1受体的变构调节剂， 确定它们如何改变配体结合;以及研究CB 1和CB 2的激动剂偏倚 通过使用具有单个G蛋白或抑制蛋白缺失的CRISPR细胞系，研究 是新颖的，核心将扩大这些研究项目超出原来的范围。因此， 将对药物滥用研究领域做出重大贡献。

Research on drug abuse and addiction requires multi-disciplinary approaches. The Biochemical Pharmacology Core provides services for researchers who do not perform biochemical pharmacology assays. The techniques provided in this Core allow interrogation of receptors associated with drugs of abuse and their signaling. Most researchers who used the Core are either behavioral scientists or chemists. The services provided by the Core are valuable to these investigators, expanded their research and have resulted in many publications. The Core also has provided preliminary data for grant applications and several NIH grants were awarded. The receptors of drugs of abuse that the Core has investigated to date include opioid receptors (MOPR, DOPR, KOPR, NOPR), cannabinoid (CB1 and CB2) receptors, nicotinic cholinergic receptors (nAChRs) and dopamine and sigma receptors. In the requested funding period, the Core will perform the following services: (1) Radioligand binding to the aforementioned receptors related to drug abuse and others when necessary; (2) Assessment of G protein-coupled receptor (GPCR) activation, including cAMP level, [35S]GTPγS binding, p44/42 MAP kinases phosphorylation, β-arrestin recruitment; (3) Determination of agonist bias for GPCRs using [35S]GTPγS binding or cAMP and β-arrestin recruitment and CRISPR cell lines; (4) Autoradiography of radioligand binding to opioid and dopamine receptors and (5) Autoradiography of opioid agonist- promoted [35S]GTPγS binding in brain sections; (6) Immunoblotting of MOPR, KOPR and phosphorylated KOPR in cultured cells and in rodent brains; (7) Internalization of MOPR, DOPR and KOPR and CB1 in cultured cells and KOPR in mouse brains; (8) Providing purified antibodies against MOPR, KOPR and phospho-KOPR to researchers; (9) Training of personnel of other laboratories. For the requested grant period, the Core will perform at least 15 collaborative projects with NIH-funded investigators across the country. The innovation lies in the proposed studies, which will address a wide variety of questions related to drug abuse, including elucidation of impact of adolescent and adult chronic exposure to morphine, cocaine or nicotine on MOPR, dopamine and/or nAChR expression and/or signaling in adults and offspring, respectively; search for KOPR agonists that do not cause KOPR phosphorylation (likely G protein-biased); effects of AMP-activated protein kinase (AMPK) deletion on expression of MOPR and nAChR; screening for allosteric modulators of CB1 receptors and determination of how they alter ligand binding; and investigation of agonist biases for CB1 and CB2 receptors by using CRISPR cell lines with either individual G proteins or arrestins deleted. The studies are novel and the Core will augment these research projects beyond the original scope. Thus, the Core will make significant contribution to the area of substance abuse research.