MOLECULAR STUDIES OF A KAPPA OPIOID RECEPTOR

阿片受体的三个主要类别，Mu，Delta和Kappa，已经被 根据它们在药理、生理和组织上的差异来定义 分发。当在体内受到刺激时，阿片受体激活一个 影响钙通道、钾离子的各种细胞内反应 通道和腺苷环化酶的活性导致许多经典的 阿片类药物中毒的快感、镇痛和生理效应 依赖。阿片受体的分子特征是 直到最近小鼠增量阿片受体的表达克隆 据报道。基于这个序列，我们和其他人开发了克隆 导致分离Kappa和Mu阿片受体的策略 CDNA。在组织中表达每种阿片受体的能力 培养使他们能够进行详细的药理和生理研究。 在之前的应用中，我们提出了R21，一种我们拥有的新型受体 克隆、编码基于关键氨基酸保守性的阿片受体 酸及其与小鼠增量阿片受体的总体同源性。拥有 证明了R21编码一种大鼠kappa阿片受体，我们现在 准备好继续进行这种受体的分子研究。在……里面 特别是，我们建议将生理学研究扩展到包括 人和大鼠kappa阿片受体与钙、钾的偶联 频道。在解剖学层面上，作为评估 我们将开始长期使用阿片剂对kappa受体表达的影响 以确定其在人脑材料中的分布。一个 阿片类药物的免疫调节作用已经被证明，最近我们 已有证据表明kappa阿片受体mRNA在小鼠体内表达 胸腺瘤细胞系。这为我们提供了一个绝佳的机会 探索阿片类药物暴露对免疫系统细胞的影响。至 更好地了解kappa阿片受体的表达是如何被控制的 将表征人类和大鼠的基因，此外还将 尝试确定可用于遗传连锁和 协会研究。最终，小鼠kappa阿片受体基因将 成为目标并被击倒。这些动物将是一个有价值的模型 评估kappa受体在过程中的作用的系统 从突触传递到行为。最后，我们克隆了一个 其序列和解剖分布表明它是一种 阿片受体基因家族的成员。我们建议继续 利用DNA序列分析来表征这种有趣的受体， 体外诱变和表达研究。

Three major classes of opioid receptors, mu, delta and kappa, have been defined based on differences in their pharmacology, physiology and tissue distribution. When stimulated in vivo the opioid receptors activate a variety of intracellular reactions that effect calcium channels, potassium channels and adenylyl cyclase activity resulting in many of the classical effects of opiate intoxication including euphoria, analgesia and physical dependence. The molecular characterization of the opioid receptors was slow until the recent expression cloning of a mouse delta opioid receptor was reported. Based on this sequence we and others developed cloning strategies that led to the isolation of both kappa and mu opioid receptor cDNAs. The ability to express each of the opioid receptors in tissue culture permits their detailed pharmacological and physiological study. In a previous application we proposed that R21, a novel receptor we had cloned, encoded an opioid receptor based on the conservation of key amino acids and its overall homology with a mouse delta opioid receptor. Having demonstrated that R21 encodes a rat kappa opioid receptor, we are now ready to proceed with the molecular studies of this receptor. In particular we propose to extend out physiological studies to include the coupling of human and rat kappa opioid receptors to calcium and potassium channels. At the anatomical level as a first step towards evaluating the effects of chronic opiate use on kappa receptor expression we will begin to characterize its distribution in human brain material. An immunomodulatory role for opioids has been demonstrated and recently we obtained evidence that kappa opioid receptor mRNA is expressed in a mouse thymoma cell line. This provides us with an excellent opportunity to explore the effects of opioid exposure on cells of the immune system. To better understand how kappa opioid receptor expression is controlled we will characterize both the human and rat genes and in addition will attempt to identify markers that can be used in genetic linkage and association studies. Eventually the mouse kappa opioid receptor gene will be targeted and knocked out. These animals will be a valuable model system in which to evaluate the kappa receptor's role in processes ranging from synaptic transmission to behavior. Finally, we have cloned a receptor whose sequence and anatomical distribution suggest that it is a member of the opioid receptor gene family. We propose to continue the characterization of this interesting receptor using DNA sequence analysis, in vitro mutagenesis and expression studies.