BRAIN CANNABINOID RECEPTOR SIGNALING AND PHARMACOLOGY

脑大麻素受体信号传导和药理学

• 批准号: 2013569
• 负责人: DEBORAH L LEWIS
• 金额: $ 16.63万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-10 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2013569
• 关键词: Cannabis; G protein; calcium channel; cannabinoid receptor; complementary DNA; guanosine triphosphate; laboratory rat; molecular cloning; neurons; neuropharmacology; potassium channel; receptor binding; receptor coupling; site directed mutagenesis; tissue /cell culture; voltage /patch clamp; voltage gated channel

DESCRIPTION: (Applicant's Abstract) Marijuana (Cannabis sativa) has had long history of use for both medicinal and recreational purposes. Since the cloning of a central nervous system cannabinoid receptor and its localization in specific brain areas; questions concerning the role of this receptor in normal brain function have arisen. Since little is known about the role of the cannabinoid receptor in normal brain function, our long term goals are to understand the basic cell biology of this receptor with the idea that this may help us understand the role of the cannabinoid receptor in learning and memory, sensory perception including pain, appetite and nausea. The first aim of our proposed research is to investigate the function of the rat brain cannabinoid receptor at the cellular electrophysiological level using a novel neuronal expression system. Preliminary experiments demonstrate that dissociated adult rat sympathetic ganglion neurons microinjected with cRNA in vitro transcribed from rat brain cannabinoid receptor cDNA express a functional cannabinoid receptor within 18 hours. Expression is both robust and reliable. The heterologously expressed rat brain cannabinoid receptor inhibits voltage-dependent calcium channel activity. This is the first demonstration of expression and functional coupling of the cloned rat brain cannabinoid receptor in an adult neuron. The ability to express an identified cannabinoid receptor and receptor mutants in normal adult mammalian neurons will facilitate identification of ion channel targets, cellular pathways and structure/function studies of the cannabinoid receptor. The second aim of our proposed research is to test the hypothesis that the cannabinoid receptor is spontaneously active and can couple to G proteins in the absence of endogenous ligands. The role of spontaneous cannabinoid receptor activity in normal brain function or drug abuse behavior is completely unknown. Our experiments will address the functional consequences of spontaneous activity of both cloned and native cannabinoid receptors. The third aim of our research is to test the functional coupling and spontaneous activity of the cannabinoid receptors in their native environment. Native cannabinoid receptors in both central and peripheral nervous system neurons will be studied using cellular electrophysiological techniques. If the cannabinoid receptor is spontaneously active in its native environment, this activity will change both normal physiology and the outcome of antagonist signaling. A spontaneously active cannabinoid receptor will have important functional consequences and negative antagonists could have potential therapeutic benefits.

描述：(申请人摘要) 大麻(大麻)有很长的药用历史 和娱乐目的。自从克隆中枢神经系统以来 大麻素受体及其在特定脑区的定位 关于这种受体在正常大脑功能中的作用的研究已经出现。 由于对大麻素受体在正常人群中的作用知之甚少 大脑功能，我们的长期目标是了解基本的细胞生物学 这可能有助于我们理解这一受体的作用 学习记忆、感觉知觉中的大麻素受体 包括疼痛、食欲和恶心。我们提出的研究的第一个目的是 是为了研究大鼠脑内大麻素受体在 一种新的神经元表达在细胞电生理水平上的应用 系统。初步实验表明，分离的成年大鼠 显微注射CRNA体外转录的交感神经节神经元 表达功能性大麻素的大鼠脑大麻素受体基因 在18小时内收到。表达既健壮又可靠。这个 异源表达的大鼠脑大麻素受体抑制 电压依赖性钙通道活动。这是第一次演示 克隆的大鼠脑大麻素的表达及功能偶联 成体神经元中的受体。表达一种身份识别的能力 正常成年哺乳动物神经元中的大麻素受体及其突变体 将有助于识别离子通道目标、细胞通路和 大麻素受体的结构/功能研究。第二个目标是 我们提出的研究是为了验证这样一种假设，即大麻素 受体是自发活跃的，在缺乏的情况下可以与G蛋白偶联 内源性配体。自发性大麻素受体的作用 大脑功能正常或药物滥用行为的活动完全 未知。我们的实验将解决以下功能后果 克隆和天然大麻素受体的自发活性。这个 我们研究的第三个目标是测试功能耦合和自发 大麻素受体在其自然环境中的活性。原生的 中枢和外周神经系统神经元中的大麻素受体 将使用细胞电生理技术进行研究。如果 大麻素受体在其天然受体中自发活跃 环境，这种活动将改变正常的生理和 拮抗剂信号传递的结果。一种自发活性的大麻素 受体会有重要的功能后果和负面影响 拮抗剂可能具有潜在的治疗益处。