MOLECULAR MECHANISMS OF PROENKEPHALIN GENE REGULATION

脑啡肽原基因调控的分子机制

• 批准号: 3212226
• 负责人: MICHAEL J COMB
• 金额: $ 13.86万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-03-01 至 1992-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3212226
• 关键词: DNA binding protein; DNA footprinting; enkephalins; gene expression; genetic enhancer element; genetic transcription; laboratory mouse; molecular cloning; neural plasticity; neuropeptides; second messengers; transcription factor

Endogenous opioids regulate not only the body's response to pain and stress but also more complex ?unctions including affect, mood, drive, and reinforcement. Control of opioid signalling occurs primarily at the level of release, precursor processing, and synthesis. Transcriptional control of opioid peptide synthesis is an important aspect of opioid signalling and may contribute to the mechanisms of narcotic tolerance, dependence, and withdrawal as well as the pathology of mood, anxiety, and other psychiatric and neurological disorders. In more general terms, an understanding of the molecular mechanisms regulating gene expression of the opioid precursor, proenkephalin, will not only improve our understanding of the processes listed above, but will also contribute to an understanding of the mechanisms regulating the synthesis of other key neuronal signalling components, including neurotransmitter receptors and ion channels. Trans-synaptic regulation of proenkephalin gene expression is an ideal model system to examine the molecular mechanisms controlling neuronal signalling plasticity because multiple forms of trans-synaptic regulation have been demonstrated in a variety of model systems and because the gene and its regulatory regions are well characterized. The primary objective of my proposed research is to determine the molecular mechanisms regulating expression of the opioid peptide precursor gene, proenkephalin, in response to synaptic inputs. Specifically, I propose an in depth analysis of the DNA binding proteins which mediate trans-synaptic activation of transcription of the proenkephalin gene via their interaction with an already identified cAMP and phorbol ester inducible DNA enhancer, as well as attempts to identify and characterize other functionally important DNA elements. In the period covered by this proposal, studies will focus on achieving a molecular description of the functional interactions between synaptic signals transduced by intracellular second messenger pathways, the DNA enhancer binding which mediate synaptic signals, and the individual DNA elements comprising the inducible enhancer. Ultimately, all the essential and modulatory components of the inducible enhancer complex will be identified and thoroughly characterized, such that the system can be functionally reconstituted, and the mechanisms of second messenger and protein kinase inducible enhancer function investigated. The proposed research is clearly of a basic nature and there is no doubt that it will be of fundamental significance for an understanding of the mechanisms regulating nerve cell signalling, opioid physiology, long-term information storage, cell growth, and differentiation.

内源性阿片类药物不仅调节身体对疼痛的反应 和压力，但也有更复杂的功能，包括影响， 心情、干劲和力量。阿片类药物信号的控制 主要发生在释放、前体处理、 和综合体。阿片肽的转录调控 合成是阿片类药物信号的一个重要方面，可能 有助于麻醉药品耐受、依赖、 和戒断以及情绪、焦虑和 其他精神和神经障碍。更一般地说 术语，对调节分子机制的理解 阿片前体前脑啡肽的基因表达不会 只是提高了我们对上面列出的过程的理解，但是 也将有助于理解这些机制 调节其他关键神经元信号的合成 成分，包括神经递质受体和离子 频道。脑啡肽原基因的跨突触调节 Expression是研究分子的理想模型系统 控制神经元信号可塑性的机制是因为 多种形式的跨突触调节已经被 在各种模型系统中进行了演示，因为基因 它的监管区域也有很好的特点。初级阶段 我提出的研究的目的是确定分子 阿片肽前体基因表达的调控机制 基因，原脑啡肽，对突触输入做出反应。 具体地说，我建议对DNA结合进行深入分析 介导跨突触激活转录的蛋白质 原脑啡肽基因通过它们与已经存在的 也鉴定了cAMP和佛波酯诱导的DNA增强子 作为试图识别和表征其他功能的尝试 重要的DNA元素。在本提案所涉期间， 研究将集中在实现分子描述的基础上 转导的突触信号之间的功能相互作用 细胞内第二信使通路，DNA增强子结合 它们传递突触信号，以及单个DNA元件 包括可诱导的增强剂。归根结底，所有必要的 并且可诱导的增强子复合体的调节成分将 被识别并彻底表征，从而使系统 可以在功能上重组，而第二种机制 信使和蛋白激酶诱导的增强子功能 调查过了。拟议的研究显然是基本性质的。 毫无疑问，它将具有根本意义 以了解神经细胞的调节机制 信号传递，阿片生理学，长期信息存储， 细胞的生长和分化。