MOLECULAR STUDIES OF A KAPPA OPIOID RECEPTOR

阿片受体的三个主要类别，μ，δ和κ，已被 根据其药理学、生理学和组织学的差异进行定义 分布 当在体内受到刺激时，阿片受体激活一种 影响钙通道、钾离子通道、 通道和腺苷酸环化酶活性导致许多经典的 阿片类药物中毒的影响，包括欣快，镇痛和身体 依赖 阿片受体的分子特征是 缓慢，直到最近表达克隆小鼠δ阿片受体， 本文报告 基于这个序列，我们和其他人开发了克隆技术， 导致kappa和mu阿片受体分离的策略 cDNA。 在组织中表达每种阿片受体的能力 培养允许对其进行详细的药理学和生理学研究。 在之前的申请中，我们提出了R21，一种新的受体， 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δ阿片受体的总体同源性。 具有 证明R21编码大鼠κ阿片受体，我们现在 准备进行这种受体的分子研究。 在 特别是，我们建议将生理学研究扩展到包括 人和大鼠κ阿片受体与钙和钾的偶联 渠道 在解剖学水平上，作为评估 长期使用阿片类药物对κ受体表达的影响我们将开始 来描述它在人脑中的分布 一个 阿片类药物的免疫调节作用已经得到证实，最近我们 获得的证据表明κ阿片受体mRNA在小鼠中表达， 胸腺瘤细胞系 这为我们提供了一个极好的机会， 探索阿片类药物暴露对免疫系统细胞的影响。 到 更好地了解κ阿片受体的表达是如何控制的， 将描述人类和大鼠的基因，此外， 尝试鉴定可用于遗传连锁的标记， 协会研究。 最终，小鼠κ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 系统中，以评估κ受体的作用，在过程中， 从突触传递到行为 最后，我们克隆了一个 受体的序列和解剖分布表明，它是一个 阿片受体基因家族成员。 我们建议继续 使用DNA序列分析表征这种感兴趣的受体， 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.