CANNABINOID RECEPTOR CHARACTERIZATION IN XENOPUS OOCYTES

描述：（申请人摘要） 大麻是一种被广泛滥用的精神活性物质，然而，某些 其药理学方面具有医药应用的潜力。 大麻素，δ-9-四氢大麻酚（δ 9-THC），是主要的 大麻的活性成分。 它产生了一个中央和 外周效应包括欣快、镇痛和免疫调节。 大麻素的作用归因于与G蛋白的相互作用 偶联受体表示为CB 1和CB 2。 大麻素配体，WIN 55，212 - 2，增强G-蛋白偶联内向整流的活性 钾通道（GIRK1和GIRK4或GIRK1/4），当通道 在非洲爪蟾卵母细胞中与人脑大麻素受体（CB 1）共表达 （X. 卵母细胞）。 目前的建议将首先侧重于评估 大麻素配体在X.卵母细胞共表达 CB1受体和GIRK1/4通道。 CB2受体并没有像 由于在体外和体内缺乏CB 2，因此作为CB 1进行了彻底研究 测定。 接下来，大麻素配体的活性将在X. 共表达CB2受体和GIRKI/4通道的卵母细胞系统。 通过定点突变改变受体蛋白中的氨基酸 已被证明是一种有效的方法， 对正常的受体功能很重要。 X.卵母细胞表达突变体 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导 转导 这些研究将增加我们对 大麻素配体-受体相互作用，并扩大我们对 大麻素受体对GIRK通道的影响 该信息可帮助 以促进更特异性的治疗剂的开发， 我们对大麻滥用的理解

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.