MOLECULAR STUDIES OF ANESTHETIC-MUSCARINIC INTERACTIONS

麻醉剂-毒蕈碱相互作用的分子研究

• 批准号: 2685053
• 负责人: MARCEL E DURIEUX
• 金额: $ 8.76万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2685053
• 关键词: CHO cells; G protein; Xenopus oocyte; analog; anesthetics; biological signal transduction; chimeric proteins; drug interactions; drug screening /evaluation; gene mutation; halothane; intermolecular interaction; isoflurane; ketamine; molecular cloning; molecular site; muscarine; muscarinic receptor; receptor coupling; site directed mutagenesis

Muscarinic acetylcholine receptors play important roles in both the peripheral and central nervous systems. They play a prominent role in the brain, and muscarinic antagonists are known to interfere with wakefulness, memory and learning. In addition, they are part of sympathetic and parasympathetic functioning, exercising profound influences on heart rate and sweating as well as on bronchial and gastrointestinal smooth muscle tone and glandular activity. Volatile anesthetics have been shown to inhibit muscarinic signaling, and some non-volatile anesthetic agents, such as ketamine and althesin, similarly appear to affect the system. These actions on muscarinic receptors could explain some of the effects as well as side-effects of the drugs. However, the studies are difficult to interpret as most times mixtures of receptor subtypes were used; single, high doses of anesthetic were studied; many studies address effects on binding rather than functional assays; and no molecular localization of the site of action was attempted. The molecular cloning of the five subtypes of muscarinic receptor now allows us to study these interactions in isolation and more detail. In this proposal, m1, m2 and m3 receptors muscarinic receptors will be expressed in Xenopus oocytes, and the effects of halothane, isoflurane, desflurane, ketamine, propofol and althesin on their functioning will be studied. Specific aims are (1) to compare these anesthetics as to their interactions with the muscarinic receptor subtypes; (2) to determine their sites of action in the muscarinic signaling cascade; (3) to localize using mutation studies the intramolecular sites of action of anesthetic drugs on muscarinic signaling; and (4) to compare the findings in Xenopus oocytes with those obtained in mammalian cells. The results of these studies will provide a better understanding of anesthetic-protein interactions in general, and of anesthetic interference with the clinically important muscarinic signaling system in particular.

毒蕈碱型乙酰胆碱受体在这两个过程中都起着重要的作用。 外周和中枢神经系统。他们在这方面发挥着重要作用， 脑，并且已知毒蕈碱拮抗剂干扰觉醒， 记忆和学习此外，他们是同情和 副交感神经功能，行使对心率的深刻影响 以及支气管和胃肠道平滑肌 音调和腺体活动。挥发性麻醉剂已被证明 抑制毒蕈碱信号传导，和一些非挥发性麻醉剂， 如氯胺酮和althesin，同样似乎影响系统。 这些对毒蕈碱受体的作用可以解释一些影响， 以及药物的副作用。然而，这些研究很难 解释为大多数时候使用受体亚型的混合物;单一， 研究了高剂量的麻醉剂;许多研究涉及对 结合而不是功能测定;并且没有分子定位， 行动的地点被尝试。 毒蕈碱受体5种亚型的分子克隆 使我们能够更详细地研究这些相互作用。在 该建议，m1，m2和m3受体毒蕈碱受体将被 在非洲爪蟾卵母细胞中表达，氟烷，异氟烷， 地氟烷、氯胺酮、异丙酚和althesin对他们的功能将是 研究了具体目的是（1）比较这些麻醉剂的 与毒蕈碱受体亚型的相互作用;（2）确定其 毒蕈碱信号级联反应的作用位点;（3）使用 突变研究麻醉药物作用的分子内位点， 毒蕈碱信号传导;（4）比较非洲爪蟾卵母细胞中的发现 与在哺乳动物细胞中获得的那些相同。这些研究的结果将 更好地了解麻醉剂-蛋白质相互作用， 一般的，和麻醉干扰的临床重要的 尤其是毒蕈碱信号系统。