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.

这一部分将继续阐明kappa的“个体发生”的分子机制 阿片受体（KOR），即在发育阶段和分化过程中产生KOR蛋白 神经元在以往的供资周期的研究已经划定了几个监管途径的控制 KOR mRNA的产生在发育阶段，主要是转录控制。到此结束 第一阶段的研究集中在KOR mRNA合成的调控上，这是KOR的一个标志。 神经发生涉及视黄酸（维生素A）和一氧化氮的信号通路，需要 KOR基因调控区的染色质重塑，以及表观遗传控制的最新发现。 重要的是，KOR的个体发生似乎是受转录后调控的， mRNA的稳定性、转运和翻译等机制。这一更新部分将侧重于 翻译机制，从我们的初步研究，已确定Netrin-1作为一个 KOR的翻译刺激因子，以及作为KOR的翻译阻遏因子的Grb 7。 两个具体目标是： 1.阐明Netrin-1/Grb 7通路激活KOR翻译的机制。我们将重点关注i） Grb 7对KOR翻译起始的调控，包括其分子和生物化学的研究 特征和功能结构域，以及由Grb 7结合的KOR RNA序列，其可以被 Netrin-1，和ii）由Grb 7靶向的特异性翻译起始步骤，包括起始因子，和 KOR mRNA的亚细胞分布和可能的环化。 2. Grb 7/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（I） 进行功能获得和丧失研究，以验证Netrin-1/Grb 7在KOR个体发育中的相关性 使用干细胞和原代神经元，和ii）检查KOR合成在神经元中的生理相关性。 神经元活动，例如在特定的疼痛回路中和在神经损伤期间或作为应激反应。