Molecular Mechanisms of Ontogenesis of K-Opioid Receptors

K-阿片受体个体发生的分子机制

• 批准号: 7612853
• 负责人: Li-Na Wei
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-30 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7612853
• 关键词: Adult; Area; Binding; Biochemical; Brain; Cell physiology; Cells; Chromatin; DNA; Development; Elements; Embryo; Epigenetic Process; Family; Funding; Genetic Transcription; Goals; Growth; Growth Factor; Heart; Ligands; Messenger RNA; Molecular; Molecular Conformation; Names; Neuronal Differentiation; Neurons; Nitric Oxide; Nucleic Acid Regulatory Sequences; Opioid; Opioid Receptor; Pain; Pathway interactions; Peptide Initiation Factors; Phase; Phenotype; Physiological; Pregnancy; Process; Production; Promoter Regions; RNA Sequences; RNA-Binding Proteins; Receptor Gene; Regulation; Regulatory Pathway; Signal Pathway; Signal Transduction; Staging; Stem cells; Transcriptional Regulation; Translation Initiation; Translational Regulation; Translations; Tretinoin; Undifferentiated; Vitamin A; biological adaptation to stress; c-myc Genes; cell type; chromatin remodeling; demethylation; early embryonic stage; human NTN1 protein; kappa opioid receptors; loss of function; mRNA Stability; nerve injury; netrin-1; neurogenesis; programs; promoter; receptor; response; stem; transcription factor

This component will continue elucidating the molecular mechanisms underlying "ontogenesis" of kappa opioid receptor (KOR), i.e. production of KOR protein during developmental stages and differentiating neurons. Studies in the previous funding cycles have delineated several regulatory pathways for the control of KOR mRNA production during developmental stages, principally transcriptional control. This concludes the first phase of studies focusing on the regulation of KOR mRNA synthesis, a hall markd of KOR neurogenesis involving signaling pathways of retinoic acid (vitamin A) and nitric oxide and requires chromatin remodeling of KOR gene regulatory regions, as well as more recent findings in epigenetic control. Importantly, ontogenesis of KOR appears to be controlled, most crucially, by post-transcriptional mechanisms such as mRNA stability, transport and translation. This renewal component will focus on translational mechanism by extending from our preliminary studies that have identified Netrin-1 as a translational stimulator for KOR, and Grb7 as a translational represser of KOR. Two specfiic aims are: 1. To elucidate the mechanism that activates KOR translation via Netrin-1/Grb7 pathway. We will focus on i) regulation of translational initiation of KOR by Grb7, including studies of its molecular and biochemical features and funcitonal domains, and KOR RNA sequences bound by Grb7 which can be activated by Netrin-1, and ii) specific translational initiation step that is targeted by Grb7 including initiation factors and subcellular distribution and possible circularization of KOR mRNA. 2. Pharmacological and physiological relevance of Grb7/Netrin-1 signaling to KOR ontogenesis. We will i) perform gain- and loss-of-function studies to validate the relevance of Netrin-1/Grb7 in KOR ontogenesis using stem cells and primary neurons, and ii) examine the physiological relevance of KOR synthesis in neuronal activity such as in a specific pain circiitry and during nerve injury or as a stress response.

该组件将继续阐明 kappa“个体发生”的分子机制 阿片受体 (KOR)，即在发育阶段和分化阶段产生 KOR 蛋白 神经元。之前的资助周期中的研究已经描绘了控制的几种监管途径 发育阶段 KOR mRNA 产生的影响，主要是转录控制。至此结束 第一阶段研究重点关注KOR mRNA合成调控，标志着KOR的殿堂 神经发生涉及视黄酸（维生素 A）和一氧化氮的信号通路，并且需要 KOR 基因调控区域的染色质重塑，以及表观遗传控制的最新发现。 重要的是，最关键的是，KOR 的个体发生似乎是由转录后控制的 mRNA 稳定性、运输和翻译等机制。该更新部分将重点关注 我们的初步研究已确定 Netrin-1 是一种翻译机制 Grb7 作为 KOR 的翻译刺激子，Grb7 作为 KOR 的翻译抑制子。 两个具体目标是： 1. 阐明通过Netrin-1/Grb7途径激活KOR翻译的机制。我们将重点关注 i) Grb7 对 KOR 翻译起始的调控，包括其分子和生化研究 特征和功能域，以及与 Grb7 结合的 KOR RNA 序列，Grb7 可以被激活 Netrin-1，和 ii) Grb7 靶向的特定翻译起始步骤，包括起始因子和 KOR mRNA 的亚细胞分布和可能的环化。 2. Grb7/Netrin-1 信号传导与 KOR 个体发生的药理学和生理学相关性。我们将我） 进行功能获得和功能丧失研究以验证 Netrin-1/Grb7 在 KOR 个体发生中的相关性 使用干细胞和原代神经元，以及 ii) 检查 KOR 合成的生理相关性 神经元活动，例如在特定的疼痛循环中和神经损伤期间或作为应激反应。