MOLECULAR MECHANISMS OF PROENKEPHALIN GENE REGULATION

脑啡肽原基因调控的分子机制

• 批准号: 3212224
• 负责人: MICHAEL J COMB
• 金额: $ 13.56万
• 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/3212224
• 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

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，还有更复杂的功能，包括情感、 情绪、动力和强化。 阿片类信号传导的控制 主要发生在释放、前体加工、 和合成。 阿片肽的转录控制 合成是阿片类信号传导的一个重要方面，可能 有助于麻醉耐受、依赖的机制， 和退缩以及情绪、焦虑和抑郁的病理学 其他精神和神经系统疾病。 更一般地说 术语，对调节分子机制的理解 阿片类药物前体脑啡肽原的基因表达不会 只会提高我们对上面列出的过程的理解，但是 也将有助于理解机制 调节其他关键神经信号的合成 成分，包括神经递质受体和离子 渠道。 脑啡肽原基因的跨突触调节 表达是检查分子的理想模型系统 控制神经信号可塑性的机制是因为 多种形式的跨突触调节 在各种模型系统中得到证明，并且因为基因 其监管区域特征明确。 初级 我提出的研究的目的是确定分子 阿片肽前体表达的调节机制 基因，脑啡肽原，响应突触输入。 具体来说，我建议对 DNA 结合进行深入分析 介导跨突触转录激活的蛋白质 脑啡肽原基因通过与已经存在的相互作用 还鉴定了 cAMP 和佛波酯诱导型 DNA 增强子 作为尝试识别和表征其他功能 重要的DNA元件。 在本提案所涉期间， 研究将集中于实现分子描述 突触信号之间的功能相互作用 细胞内第二信使途径，DNA增强子结合 介导突触信号和单个 DNA 元件 包含诱导增强子。 最终，所有必需的 和诱导增强子复合物的调节成分将 被识别并彻底表征，以便系统 可以在功能上重建，并且第二个的机制 信使和蛋白激酶诱导增强子功能 调查了。 拟议的研究显然具有基础性质 毫无疑问，这将具有根本性意义 了解调节神经细胞的机制 信号传导、阿片类生理学、长期信息存储、 细胞生长、分化。