MU OPIOID RECEPTOR GENE IN HEROIN ADDICTS

海洛因成瘾(二乙酰吗啡)是一个主要的社会问题， 影响了美国100多万人。在身体里， 在大脑中，海洛因被水解成吗啡，吗啡作用于u阿片类药物 受体并产生愉悦效应，从而赋予 增强药物的性能，为开发 上瘾。海洛因成瘾可以通过治疗来控制，主要是 美沙酮维持，但海洛因成瘾的生物学基础， 关于Mu受体的遗传多样性及其影响 受体功能的多样性。克隆了人类Mu阿片类药物 受体，现在可以研究它们之间的序列差异。 不同个体及其与受体变化的关系 功能。 这一建议的假设是Mu中的dna序列变异 受体基因存在于人类体内，这种自然发生的多态现象 影响受体功能对吗啡的反应。为了测试这一点 假设，血液样本将从海洛因成瘾者和正常人群中采集 志愿者建立一个包含DNA和可存活DNA的存储库 淋巴细胞系，储存重复样本，其中一组设在印第安纳 一所大学，一所洛克菲勒大学。这些基因的多态现象 个体将通过扩增Mu受体编码来确定 用聚合酶链式反应进行测序，并筛选序列变异。一种相关性 多态和受体功能变化之间的关系将通过以下方式进行研究 突变Mu受体基因以匹配多态。这些突变 受体将在培养的细胞中表达，并测试其 在配基显带、抑制腺苷环化酶活性方面的作用，以及 钾通道的激活。吗啡作用的时间进程将 研究急性刺激、慢性暴露和急性戒断。 此外，还将研究蛋白激酶对受体功能的调节。 作为药物引起细胞变化的分子机制。 这是一种研究其生物学意义的新方法。 麻醉剂受体。通过使用一组控制良好的受试者。使用 关于健康和药物滥用史的详细信息，详细的分子 对每个受试者的Mu受体基因的分析应该会产生有价值的 关于Mu受体基因和功能多样性的信息 提供对其结构-功能关系的直接评估。 此外，这种方法也可以很容易地应用于其他蛋白质。 参与滥用药物的生理途径，产生重要的 关于这些蛋白质的生物学机制的信息，以及 为药物成瘾的生物学基础提供了广阔的视角。

Addiction to heroin (diacetylmorphine) is a major social problem, affecting over a million people in the United States. In the body and brain, heroin is hydrolyzed to morphine, which acts at the mu opioid receptor and results in an euphoric effect, thus conferring the reinforcing properties of the drug and contributing to the development of addiction. Heroin addiction can be managed through treatment, primarily methadone maintenance, but the biological basis of heroin addiction, with respect to the genetic diversity of the mu receptor and the impact of such diversity on receptor function. With the cloning of the human mu opioid receptor, it is possible now to study the sequence variations among different individuals and correlate them with changes in receptor function. The hypothesis for this proposal is that DNA sequence variations in the mu receptor gene exist in humans, and such natural occurring polymorphisms affect the receptor function in response to morphine. To test this hypothesis, blood samples will be collected from heroin addicts and normal volunteers to establish a repository that contains DNA and viable lymphocyte cell lines, storing duplicate samples with one set at Indiana University and one at Rockefeller University. Polymorphisms in these individuals will be determined by amplifying the mu receptor coding sequence using PCR and screening for sequence variations. A correlation between polymorphisms and changes in receptor function will be examined by mutating the mu receptor cDNA to match the polymorphisms. These mutated receptors will be expressed in cultured cells, and tested for their functions in ligand banding, inhibition of adenylyl cyclase activity, and activation of a potassium channel. The time course of morphine effect will be studied upon acute stimulation, chronic exposure, and acute withdrawal. Also, modulation of receptor function by protein kinases will be examined as a molecular mechanism for drug-induced changes in the cell. This is a novel approach for studying the biological significance of narcotic receptors. By using a well-controlled group of subjects. with detailed information on health and drug abuse history, detailed molecular analysis of each subject's mu receptor gene should yield valuable information on the genetic and functional diversity of the mu receptor and provide a direct assessment of its structure-function relationship. Furthermore, this approach can be readily applied to other proteins involved in the physiological pathways of abused drugs, yielding important information regarding the biological mechanisms for these proteins, and provide a broad perspective on the biological basis of drug addiction.