Molecular Mechanisms of Ontogenesis of K-Opioid Receptors

K-阿片受体个体发生的分子机制

基本信息

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
项目类别：
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

项目摘要

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.

该成分将继续阐明Kappa“个体发生”的分子机制阿片受体（KO），即在发育阶段生产Kor蛋白质和区分神经元。先前的资金周期的研究已经描述了对照的几种调节途径发育阶段的KOR mRNA产生，主要是转录控制。这是总结研究的第一阶段，重点介绍了Kor mRNA合成的调节，这是Kor的Hall Markd 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基因调节区域的染色质重塑，以及表观遗传控制中的最新发现。重要的是，最关键的是，Kor的个体发生似乎是通过转录后控制的 MRNA稳定性，运输和翻译等机制。此更新组件将重点放在通过从我们的初步研究中延长的转化机制，这些研究已将Netrin-1确定为一个 Kor的翻译刺激器和GRB7作为KOR的翻译压抑者。两个特定目标是： 1。阐明通过Netrin-1/grb7途径激活KOR翻译的机制。我们将专注于我） GRB7对KOR的翻译启动的调节，包括对其分子和生化的研究特征和funcitonal域，以及由GRB7绑定的Kor RNA序列，可以通过 Netrin-1和ii）GRB7针对的特定翻译起始步骤，包括启动因子和 kor mRNA的亚细胞分布和可能的循环。 2。grb7/netrin-1信号与kor非正式发生的药理和生理相关性。我们会）执行功能丧失研究，以验证Netrin-1/grb7在KON ONTTOGENESID中的相关性使用干细胞和原发性神经元，ii）检查KOR合成在神经元活性，例如在特定的疼痛循环中，神经损伤或作为压力反应。