Agonist-Dependent Signaling and Post-Signaling Events of DOR

DOR 的激动剂依赖性信号传导和信号后事件

基本信息

批准号：
7612856
负责人：
HORACE LOH
金额：
$ 17.83万
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/7612856
关键词：
ADRBK2 gene Accounting Adverse effects Affect Agonist Arrestin Arrestins Beta-Adrenergic Receptor Kinase 2 Biological Assay Cell Nucleus Cell model Cells Chronic Clinical Complex Corpus striatum structure Cytosol Data Dependence Development Drug Interactions Elements Embryo Endopeptidases Etorphine Event Fentanyl Fibroblasts G-Protein-Coupled Receptors G-protein-coupled receptor kinase 3 Goals In Vitro Knockout Mice Mediating Modeling Molecular Monitor Morphine Mus N-Methyl-D-Aspartate Receptors N-Methylaspartate Neuroblastoma Neurons Neurotransmitter Receptor Nuclear Opioid Opioid Analgesics Opioid Receptor Pain management Pathway interactions Peptide Hydrolases Peptides Pharmaceutical Preparations Phosphotransferases Progress Reports Protein Isoforms Protein Kinase Receptor Signaling Role Signal Pathway Signal Transduction Small Interfering RNA Specificity Spinal cord posterior horn Subfamily lentivirinae System Transcript Work arrestin B base cell growth regulation desensitization design in vivo inhibitor/antagonist member novel protein protein interaction receptor receptors for activated C kinase research study small hairpin RNA transcription factor

项目摘要

The molecular mechanism for morphine tolerance has not been firmly established yet. The current model of [j-opioid receptor (MOR) desensitization via the &-Arrestin pathway cannot account for the numerous observations that other neurotransmitter receptor activities, such as NMDA, could contribute to morphine tolerance. The activity of other opioid receptors, such as the 6-opioid receptor (DOR), could be implicated in morphine tolerance development also. Since morphine can activate and desensitize DOR during prolonged treatment, our working hypothesis is that the post-signaling events occurring within the DOR-containing neurons during morphine treatment contribute to tolerance development. Our working hypothesis also is that morphine has post-signaling events distinct from those of other opioid agonists. In order to demonstrate these hypotheses, agonist-dependent signaling events will be established for morphine activation of DOR. In our studies with MOR signaling, we have demonstrated that morphine differs from other agonists in its pathway to activate ERK1/2. Agonists such as etorphine activate ERK1/2 via the B-Arrestin-dependent pathway, while morphine activates ERK1/2 via the PKC-dependent pathway. This divergent activation results in differential translocation of the activated ERK1/2 and the transcripts produced. Therefore, the signaling pathway and the post-signaling events of morphine in cell models expressing DOR will be established. The possible involvement of the PKC-dependent pathway on morphine-mediated DOR activation of ERK1/2 will be studied. The specific PKC subtypes involved will be defined. The reasons for the differences among agonists in selecting a pathway will be investigated by monitoring protein-protein interactions using a novel protease assay system. Parallel studies will be conducted with primary neuronal cultures. The blockade of specific PKC subtypes in DOR-expressing neurons on in vivo morphine tolerance development will be explored. By selectively inactivating the morphine signaling pathway, and subsequently its post-signaling events in DOR-containing neurons, possible blockade of morphine tolerance without altering morphine activities in MOR containing neuron could be accomplished.

吗啡耐受的分子机制尚未明确。目前的型号为 [j-阿片受体 (MOR) 通过 &-Arrestin 途径脱敏不能解释许多观察到其他神经递质受体活性（例如 NMDA）可能有助于吗啡宽容。其他阿片受体的活性，例如 6-阿片受体 (DOR)，可能与吗啡耐受性也发生。由于吗啡可以在长时间使用时激活 DOR 并使 DOR 脱敏治疗中，我们的工作假设是，包含 DOR 的信号后事件发生在吗啡治疗期间的神经元有助于耐受性的发展。我们的工作假设也是吗啡具有与其他阿片类激动剂不同的后信号事件。为了证明这些假设将建立吗啡激活 DOR 的激动剂依赖性信号传导事件。在我们对 MOR 信号传导的研究表明，吗啡与其他激动剂的不同之处在于激活 ERK1/2 的途径。埃托啡等激动剂通过 B-Arrestin 依赖性激活 ERK1/2 途径，而吗啡通过 PKC 依赖性途径激活 ERK1/2。这种不同的激活结果激活的 ERK1/2 和产生的转录物的差异易位。因此，信令将建立表达 DOR 的细胞模型中吗啡的途径和信号后事件。这 PKC 依赖性途径可能参与吗啡介导的 ERK1/2 DOR 激活被研究。所涉及的具体 PKC 亚型将被定义。造成差异的原因将通过使用一种新的方法监测蛋白质-蛋白质相互作用来研究选择途径的激动剂蛋白酶测定系统。将利用原代神经元培养物进行平行研究。的封锁 DOR表达神经元中特定的PKC亚型对体内吗啡耐受发展的影响将是探索过。通过选择性地灭活吗啡信号通路及其后信号传导含有 DOR 的神经元中的事件，可能在不改变吗啡的情况下阻断吗啡耐受可以完成含有神经元的 MOR 的活动。