Studies of mouse kappa opioid receptor gene regulation

描述(由申请人提供)： 这项建议的目的是继续我们正在进行的调查 小鼠kappa阿片受体基因表达的调控机制 它目前由一架R01提供资金。遵循相同的原则 如原项目所建议的方向，本次续展申请进一步 有助于理解阿片受体调控中的基本问题 通过组织培养和转基因小鼠模型进行表达。它是 已知阿片类药物在体内具有广泛的药理和生理作用 动物。像许多其他药物一样，阿片类药物通过特定的受体作用于 目标单元格。它们在成人神经系统中的表达非常有限 在发育阶段表明其特异性和水平 阿片受体的表达必须受到严格的调控，并引发一个重要的 在动态平衡状态下对其表达的控制问题。在 在之前的颁奖期间，我们选择了KOR基因作为解决这一问题的模式 问题，我们的数据表明KOR基因受到多层次的调控 涉及正转录调控和负转录调控的回路 机制和几个转录后事件，如选择性剪接 还有翻译。负性转录调控是通过 维生素A激素(维甲酸)的作用，而选择性剪接 产生具有不同RNA稳定性的KOR mRNA亚型，并翻译 效率，并可差异化地转运到神经元胞体和 纤维。 在与原始项目相同的方向上，我们将继续解决我们的 KOR基因维持动态平衡的基本假说 整合不同层级的监管事项。在转录杂志上 在干细胞分化过程中，KOR在两个阶段受到调节， 一种内分泌因子维甲酸所产生的最初的负面影响， 其次是未知的积极机制，以重新激活神经元中的KOR基因。 最终，我们将了解到：a)KOR基因的分子/遗传学基础 转录调控，特别是在染色质的背景下，b)KOR 差异mRNA稳定性的基因转录后调控 和运输，以及c)KOR基因调控机制从细胞延伸而来 培养动物(转基因小鼠)。此次续订的一个重要特点 提议是我们将在一个更具生理学意义的 通过研究染色质水平上的调节事件，也可以改变细胞的状态。 此外，转基因小鼠模型的应用对于 了解与以下相关的药理学问题的遗传基础 阿片受体系统。

DESCRIPTION (provided by applicant): The purpose of this proposal is to continue our ongoing investigation on the regulatory mechanisms of the mouse kappa opioid receptor (KOR) gene expression which is currently funded by an R01. Following the same direction as proposed in the original project, this renewal application further serves to understand the fundamental problems in the control of opioid receptor expression, by using both tissue culture and transgenic mouse models. It is known that opiates exert extensive pharmacological and physiological effects in animals. Like many other drugs, opioids act through specific receptors on target cells. Their very restricted expression in the adult nervous system and during developmental stages suggests that the specificity and the level of opioid receptor expression must be tightly regulated, and raises an important question of the control for their expression in a homeostatic state. In the previous award period, we have selected the KOR gene as a model to address this question, and our data indicate that KOR gene is under a multi-level regulatory circuit that involves positive and negative transcriptional regulatory mechanisms and several post-transcriptional events such as alternative splicing and translation. The negative transcriptional control is mediated through the action of vitamin A hormone (the retinoic acid), whereas alternative splicing generates KOR mRNA isoforms that have distinct RNA stability, and translation efficiency, and can be differentially transported to neuron cell bodies and fibers. In the same direction as the original project, we will continue to address our principle hypothesis that KOR gene is maintained at a homeostatic state by integrating different levels of regulatory events. At the transcriptional level, KOR is regulated in two phases during stem cell differentiation, with an initial negative effect exerted by an endocrine factor, the retinoic acid, followed by unknown positive mechanisms to reactivate KOR gene in neurons. Ultimately, we will learn: a) the molecular/genetic basis of KOR gene transcriptional regulation, particularly in the context of chromatin, b) KOR gene post-transcriptional control with regard to differential mRNA stability and transport, and c) KOR gene regulatory mechanisms extended from cell cultures to animals (transgenic mice). One important feature of this renewal proposal is that we will test our hypotheses in a more physiologically relevant condition by also studying the regulatory events at the chromatin level. Additionally, the application of transgenic mouse models is essential for understanding the genetic basis of pharmacological problems that are related to the opioid receptor system.