Studies of the mouse kappa opioid receptor gene

小鼠κ阿片受体基因的研究

• 批准号: 6864823
• 负责人: Li-Na Wei
• 金额: $ 12.36万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6864823
• 关键词: gene expression; genetic promoter element; genetic regulation; genetic transcription; genetically modified animals; intracellular transport; laboratory mouse; messenger RNA; neurons; neuropharmacology; opioid receptor; receptor expression; retinoate; tissue /cell culture; training

DESCRIPTION: (provided by the applicant) The purpose of this K02 application is to seek salary support for scientific career development of the PI at the Dept. Pharmacology of Univ. Minnesota Medical School. The PI was trained in molecular biology and genetics with special focus on interaction of hormone nuclear receptors and transcription factors. She has initiated a research project (DA-11190) aimed to study regulatory mechanisms for kappa opioid receptor (KOR) gene regulation. In order to develop her sceintific career along the line of drug abuse research and perform several experiments described in this project, she plans to devote more time into her research and receive further training in developmental neurobiology and opioid pharmacology. The project proposed is based upon the currently funded R01 to examine several important regulatory mechanisms underlying KOR gene expression. Three aims are proposed to address two fundamental problems in KOR gene regulation: transcription and post-transcriptional mechanisms for controlling KOR expression. Three aims are: 1)To Study Transcriptional Regulation of Mouse KOR Gene, 2) To Study Post-transcriptional Regulation of Mouse KOR Expression and 3) Transgenic Mouse Models to Study KOR Ontogeny and Regulation. The first aim will address transcriptional control by focusing on a retinoic acid (RA)-regulated silencer element located in intron 1 of the gene, and the positive mechanisms regulating promoters 1 and 2. The mechanisms underlying histone deacetylation on KOR promoters as a result of retinoid depletion will be addressed. Protein factors responsible for the activation of KOR gene in KOR neurons will be identified. An embryonic cell line P19 that is capable of differentiation into various cell lineages including KOR neurons and expresses KOR in an RA-regulated manner will be used. In the second aim, mRNA transport and protein translation efficiency will be examined in relation to the cell- and tissue type-specificity of KOR expression. The information gathered from aims 1 and 2 deals with KOR regulation in vitro. The third aim is to confirm and correct the information generated from in vitro models by using transgenic mouse models. These genetic studies will answer whether and how pharmacological problems, such as opioid addiction and tolerance, may be related to the endocrine system such as retinoid status and the homeostatic control of opioid receptor gene activity. The granting of this award will enable the PI to devote more time into her research projects and take leave of absence to receive training. This award is essential for the PI?s career development as a more productive and contributing scientist in the field of drug abuse research.

描述：（由申请人提供） 本次K02申请的目的是为科研人员寻求薪资支持 大学药理学系PI的职业发展明尼苏达州 医学院。 PI 接受过分子生物学和遗传学方面的培训 特别关注激素核受体和转录的相互作用 因素。她发起了一个研究项目（DA-11190），旨在研究 kappa 阿片受体 (KOR) 基因调控的调控机制。为了 沿着药物滥用研究的方向发展她的科学事业 进行该项目中描述的多项实验，她计划投入更多 投入时间进行研究并接受进一步的发展培训 神经生物学和阿片类药物药理学。拟议的项目是基于 目前资助R01来研究几个重要的监管机制 潜在的 KOR 基因表达。提出三个目标来解决两个问题 KOR基因调控的基本问题：转录和 控制 KOR 表达的转录后机制。三个目标是： 1）研究小鼠KOR基因的转录调控，2）研究 小鼠 KOR 表达的转录后调控和 3) 转基因小鼠 研究 KOR 个体发育和调控的模型。第一个目标将解决 通过关注视黄酸（RA）调节的沉默子进行转录控制 位于该基因内含子1的元件及其正向调节机制 启动子 1 和 2。KOR 上组蛋白脱乙酰化的机制 将解决因类视黄醇耗尽而导致的启动子问题。蛋白质因子 将鉴定负责 KOR 神经元中 KOR 基因激活的因素。 能够分化成多种细胞的胚胎细胞系P19 包括 KOR 神经元的谱系并以 RA 调节的方式表达 KOR 将 被使用。第二个目标，mRNA运输和蛋白质翻译效率 将根据 KOR 的细胞和组织类型特异性进行检查 表达。从目标 1 和 2 收集的信息涉及 KOR 体外调节。第三个目的是确认和更正信息 使用转基因小鼠模型从体外模型产生。这些遗传 研究将回答阿片类药物是否以及如何药理学问题 成瘾性和耐受性，可能与内分泌系统有关，如 类维生素A状态和阿片受体基因活性的稳态控制。 该奖项的授予将使PI能够投入更多的时间 研究项目并请假接受培训。这个奖项是 对于 PI 的职业发展至关重要，使其成为更有生产力和贡献的人 药物滥用研究领域的科学家。