MOR PHosphorylation in Opiod Tolerance and Dependence

描述(由申请人提供)： 要了解药物成瘾的生物学基础，遗传学和神经科学是两个非常重要的因素。朝着这个方向，PL的研究开发和培训目标是追求基于基因打靶技术和活体动物模型的新方法的开发和应用。此外，与药物滥用研究相关的教育和服务活动已纳入建议，以进一步促进律师的职业发展。激动剂诱导的阿片受体磷酸化被认为是一个重要的受体调节过程，促进了急性受体脱敏，触发了受体的内化和再循环，参与了耐受的形成。然而，这还没有在神经系统和整个动物中得到证实。因此，提出的研究计划旨在验证这样的假设，即MOR的磷酸化是激动剂依赖的，也是区域或细胞特异性的，以及受体的微构象和细胞信号成分影响磷酸化的程度和体内差异耐受和依赖的发展。以原代培养的DRG神经元为模型，PI建议检测DRG神经元中MOR的磷酸化模式，并利用磷酸化缺失的突变体受体来评估其在脱敏和其他受体调节过程中的作用。PI还建议确定受体二聚化对MOR磷酸化的影响，探索受体二聚化对受体磷酸化的潜在区域差异和阿片类药物耐受发展的可能贡献。PI进一步建议通过使用敲入方法产生MOR磷酸化缺陷突变小鼠来改变动物的MOR磷酸化状态，从而在体内检验这一假说。这项提议的一个意义在于，这项研究旨在了解阿片类药物在生理相关环境中的作用机制。

DESCRIPTION (provided by applicant): To understand the biological basis of drug addiction, genetics and neuroscience are two very important elements. Towards this direction, the Pl's research development and training goals are to pursue the development and application of new methodologies based on gene targeting technology and in vivo animal models. In addition, drug abuse research related educational and severice activities have been incorporated into the proposal to further enhance Pl's career development. Agonist-induced opioid receptor phosphorylation is believed to be an important receptor regulatory process that promotes acute receptor desensitization, triggers internalization and recycling of the receptors, and participates the development of tolerance. However, this has not been demonstrated in neuronal systems and whole animals. Therefore, the proposed research plan is aimed to test the hypothesis that MOR phosphorylation is agonist-dependent as well as region or cell specific, and that the receptor's microconformation and cellular signaling components influence the extent of phosphorylation and the development of differential tolerance and dependence in vivo. Using primary culture of DRG neurons as a model, the PI proposes to examine the MOR phosphorylation pattern in DRG neuron and to evaluate its contribution to desensitization in and other receptor regulatory processes by using phosphorylation-deficient mutant receptor. The PI also proposes to determine the effect of receptor dimerization on MOR phosphorylation, exploring the possible contributions of receptor dimerization to the potential regional variation in receptor phosphorylation and the development of opioid tolerance. The PI further proposes to test the hypothesis in vivo by modifying the MOR phosphorylation status in animal through generating MOR phosphorylation-deficient mutant mice using a knock-in approach. A significance of the proposal is that the study is designed to understand the mechanism of opioid action in a physiologically relevant setting.