MOLECULAR MECHANISMS OF PROENKEPHALIN GENE REGULATION

脑啡肽原基因调控的分子机制

• 批准号: 3069538
• 负责人: MICHAEL J COMB
• 金额: $ 5.92万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3069538
• 关键词: DNA footprinting; behavioral genetics; biological signal transduction; calcium flux; enkephalins; gene expression; genetic enhancer element; genetic regulation; genetic transcription; laboratory mouse; molecular cloning; neural transmission; nucleic acid probes; nucleoproteins; oxidative phosphorylation; phorbols; polymerase chain reaction; protein kinase; protein purification; protein sequence; protein structure function; second messengers; site directed mutagenesis; synapses; tissue /cell culture; transcription factor; transfection

My long range goals are to develop an understanding of how neural activity regulates gene expression, and to ultimately understand how these processes contribute to environmental and drug induced stable/adaptive changes in the nervous system. A better understanding of theses processes will help us understand the adaptive physiologic changes underlying drug addiction, tolerance, withdrawal, and drug-seeking behaviors. The molecular mechanisms underlying learning, memory, long-term effects of addictive drugs, and other stable/adaptive changes in the nervous system are largely unknown. With the discovery that neurons transduce environmental inputs into changes in gene expression, the hypothesis has emerged that trans-synaptic regulation of gene expression may induce the biochemical changes underlying many forms of neural signaling plasticity. To understand these processes it is necessary to identify both the pathways and mechanisms mediating the effects of learning, psychotropic drugs, and other synaptically mediated processes on gene expression. Neural regulation of the opioid precursor, proenkephalin, is an excellent model system to study mechanisms underlying synaptic regulation of gene expression because: i) transcription is both activated and repressed by synaptic signals, ii) in the brain, events such as electrical stimulation which induce LTP also induce dramatic and long lasting changes in proenkephalin gene expression, iii) the gene products, enkephalins, are important signaling molecules regulating diverse neural pathways including analgesia, reward and motivational states, and hormone release. The primary objective of this research proposal is to identify and characterize nucleoprotein complexes which regulate proenkephalin gene expression and to determine how these nucleoprotein complexes mediate synaptic regulation of proenkephalin transcription via their interaction with a well characterized second messenger inducible DNA enhancer. In the period covered by this proposal, studies will focus on defining components of ENKTF-1 and Fos/Jun nucleoprotein complexes which mediate synaptic regulation of proenkephalin transcription. In addition their functional and biochemical interactions with the proenkephalin inducible DNA enhancer and signals transmitted through intracellular signaling pathways will be investigated. Because the transcription factors described above are activated by both neurotransmitters and pharmacologic agents they can be considered to be important targets of drug action. Progress in understanding these mechanisms should provide insight into the pathogenesis and treatment of many disorders affecting the nervous system.

我的长期目标是了解神经活动 调节基因表达，并最终了解这些过程 有助于环境和药物诱导的稳定/适应性变化， 神经系统 更好地理解这些过程将有助于我们 了解药物成瘾背后的适应性生理变化， 耐受性、戒断和寻求药物的行为。 学习记忆的分子机制， 成瘾性药物和神经系统中的其他稳定/适应性变化 大部分都是未知的。 随着发现神经元 环境输入到基因表达的变化，该假设有 研究表明，基因表达的跨突触调节可能会诱导基因表达。 许多形式的神经信号可塑性的生化变化。 为了理解这些过程，有必要确定这两种途径 以及调节学习效果的机制，精神药物， 其他突触介导的基因表达过程。 神经 阿片样物质前体前脑啡肽的调节是一个很好的模型， 研究基因突触调控机制系统 表达，因为：i）转录被激活和抑制， 突触信号，ii）在大脑中，事件，如电刺激 这些诱导LTP的蛋白质也诱导了 脑啡肽原基因表达，iii）基因产物，脑啡肽， 重要的信号分子调节不同的神经通路，包括 镇痛、奖励和激励状态，以及激素释放。 的 本研究提案的主要目标是识别和表征 调节脑啡肽原基因表达的核蛋白复合物， 确定这些核蛋白复合物如何介导突触调节 脑啡肽原转录通过它们与一个充分表征的 第二信使可诱导的DNA增强子。 所涉期间 根据提议，研究将侧重于确定ENKTF-1和Fos/Jun的组成部分， 介导脑啡肽原突触调节的核蛋白复合物 转录。 此外，它们的功能和生物化学相互作用 与前脑啡肽诱导的DNA增强子和信号传输 通过细胞内信号通路将被研究。 因为 上述转录因子被两种细胞激活， 神经递质和药理剂，它们可以被认为是 毒品的重要目标。 了解这些进展 机制应该提供深入了解的发病机制和治疗 许多影响神经系统的疾病。