CANNABINOID RECEPTOR CHARACTERIZATION IN XENOPUS OOCYTES

描述：(申请人摘要) 然而，大麻是一种被广泛滥用的精神活性物质，然而， 其药理方面具有潜在的医药应用价值。 大麻素，三角洲-9-四氢大麻酚(Delta9-THC)是主要的 大麻的有效成分。它产生了多个中央和 外周效应包括快感、止痛和免疫调节。 大麻素的作用被归因于与G蛋白的相互作用 偶联受体表示CB1和CB2。大麻素配体，Win 55,212-2，增强G蛋白偶联内向整流活性 钾通道(GIRK1和GIRK4或GIRK1/4) 与人脑大麻素受体(CB1)在非洲爪哇卵母细胞中的共表达 (X.卵母细胞)。目前的提案将首先集中于评估 大麻素配体在共表达大麻素的X.卵母细胞系统中的活性 CB1受体和GIRK1/4通道。CB2受体还没有被当作 由于在体外和体内缺乏CB2，因此被深入研究为CB1 化验。接下来，将用X来比较大麻素配体的活性。 共表达CB2受体和GIRKI/4通道的卵母细胞系统 用定点突变技术改变受体蛋白中的氨基酸 已经证明是一种有效的方法，可以用来绘制 对正常的受体功能很重要。X.表达突变的卵母细胞 大麻素受体结构和GIRK1/4将用于确定 CB1和CB2受体结构域参与配体识别和信号传递 转导。这些研究将增加我们对……本质的认识。 大麻素配体-受体相互作用和拓宽我们对 大麻素受体对GIRK通道的影响。这些信息可能会有所帮助 推动更具特异性的治疗剂的发展和进步 我们对大麻是一种滥用毒品的理解。

DESCRIPTION: (Applicant's Abstract) Marijuana is a widely abused psychoactive substance, however, certain aspects of its pharmacology have the potential for medicinal applications. The cannabinoid, delta-9-tetrahydrocannabinol (delta9-THC), is the principal active constituent of marijuana. It produces a multiplicity of central and peripheral effects including euphoria, analgesia, and immunomodulation. Cannabinoid effects have been attributed to interactions with the G-protein coupled receptors denoted CB1 and CB2. The cannabinoid ligand, WIN 55,212-2, enhances the activity of G-protein coupled inwardly rectifying potassium channels (GIRK1 and GIRK4 or GIRK1/4) when the channels are co-expressed with human brain cannabinoid receptor (CB1) in Xenopus oocytes (X. oocytes). The current proposal will first focus on evaluating the activities of cannabinoid ligands in a X. oocyte system co-expressing the CB1 receptor and GIRK1/4 channels. The CB2 receptor has not been as thoroughly studied as CB1 due to the lack of CB2 in vitro and in vivo assays. Next the activities of cannabinoid ligands will be compared in a X. oocyte system co-expressing the CB2 receptor and GIRKI/4 channels. Alterations of amino acids in receptor proteins by site directed mutagenesis has proven to be an effective method with which to map regions that are important for normal receptor function. X. oocytes expressing mutant cannabinoid receptor constructs and GIRK1/4 will be used to determine which CB1 and CB2 receptor domains are involved in ligand recognition and signal transduction. These studies will increase our knowledge of the nature of cannabinoid ligand-receptor interaction and broaden our understanding of cannabinoid receptor influence on GIRK channels. This information may help to promote the development of more specific therapeutic agents and advance our understanding of marijuana as a drug of abuse.