PSYCHOTROPIC DRUG RESPONSIVE TRANSCRIPTION FACTORS

描述(申请人摘要)： 吸毒成瘾是美国的主要公共卫生问题之一 并在很大程度上抵制目前可用的预防和 治疗策略。因此，近年来， 对数量众多的实验室感兴趣，包括我自己的，在定义 细胞内信号转导途径调节和调节长期 神经元对多种生理和药理学的适应 刺激物，包括滥用药物。作为这项持续努力的一部分，我 请求续签我的K02奖。这次续约将使我能够获得 我在分子生物学方法方面的进一步专业知识 以前的研究。 本提案中概述的研究的具体目标侧重于 界定即刻早期的Egr家族调控的新方面 基因转录因子。特别是，我们在 初步研究表明，有明显的急性期和延迟期 由Egr反应元件介导的转录反应。作为 延迟阶段可能与理解长期 滥用药物对神经功能的影响，我们计划进行 实验的目的是：1)确定经历过的Egr家庭成员 Egr家族表达的“延迟”波，以及2)如何决定这两个 Egr家族成员表达的不同阶段受重复或慢性影响 刺激。此外，在旨在确定 在延迟阶段表达的EGR家族成员，我们已经获得 有证据表明其中一种蛋白Egr-3在体内被磷酸化。就像小事一样 了解Egr家族成员是如何被磷酸化调控的，我们计划 3)绘制Egr-3在基础条件下的磷酸化位点图， 在第二信使通路被激活后，以及4)评估 磷酸化对其转录调控活性的影响。 通过开展以下工作获得进一步的分子生物学方法专业知识 这项提案中概述的研究将为 继续进行未来的研究，旨在定义信号通路， 调节神经元的可塑性和精神药物的作用。收盘 约翰斯大学神经科学系成员之间的互动 霍普金斯大学，其中包括分子神经生物学的领先专家，提供了 为我的科学发展提供了理想的环境。

DESCRIPTION (Applicant's Abstract): Drug addiction is one of the major public health problems in the United states and is largely resistant to currently available prevention and treatment strategies. Accordingly, in recent years, there has been intense interest in numberous laboratories, including my own, in defining the intracellular signalling pthways that regulate and mediate the long-term adaptation of neurons to a wide variety of physiological and pharmacological stimuli, including drugs of abuse. As part of this ongoing effort, I am requesting renewal of my K02 award. This renewal will enable me to gain further expertise in molecular biological approaches I have employed in previous studies. The specific aims of the research outlined in this proposal focus on defining novel aspects of regulation of the Egr family of immediate early gene transcripiton factors. In particular, we have obtained evidence in preliminary studies that there are distinct acute and delayed phases to the transcriptional responses mediated by the Egr response element. As the delayed phase may be especially relevant to understanding the long-term effects of drugs of abuse on neuronal funciton, we plan to conduct experiments aimed at: 1) defining the Egr family members experessed durng the "delayed" wave of Egr family expression, and 2) determine how these two phases of Egr family member expression are influenced by repeated or chronic stimulation. In addition, in preliminary studies aimed at identifying the Egr family members expresed during the delayed phase, we have obtained evidence that one of these, Egr-3, is phosphorylated in vivo. As little is known about how Egr family members are regulated by phosphorylation, we plan to 3) map the sites of phosphorylation of Egr-3 under basal conditions and following activation of second messenger pathways, and 4) to assess the effects of phosphorylation on its transcriptional regulatory activity. Acquiring further expertise in molecular biological approaches by carrying out the studies outlined in this proposal will provide a firm foundation for pursuing future studies aimed at defining the signalling pathways that regulate neuronal plasticity and psychotropic drug actions. The close interaction among members of the Department of Neuroscience at Johns Hopkins, which include leading experts in molecular neurobiology, provides an ideal environment for my scientific development.