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

• 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
• 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

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.

吗啡耐受性的分子机制尚未牢固确定。当前模型 [通过＆-arrestin途径脱敏的J-阿片类受体（MOR）无法解释众多 观察其他神经递质受体活性（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 吗啡耐受性的发展。由于吗啡可以在延长期间激活和脱敏DOR 治疗，我们的工作假设是在含DOR中发生的信号事件 吗啡治疗期间的神经元有助于耐受性发展。我们的工作假设也是 吗啡的信号事件与其他阿片类药物激动剂的事件不同。为了证明 这些假设，依赖激动剂的信号事件将建立用于DOR的吗啡激活。在 我们对MOR信号传导的研究，我们已经证明吗啡与其他激动剂的不同 激活ERK1/2的途径。像e丁碱这样的激动剂通过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 在激活的ERK1/2的差异转运和产生的转录本。因此，信号传导 将建立表示DOR细胞模型中吗啡的途径和信号事件。这 PKC依赖性途径可能参与吗啡介导的ERK1/2的DOR激活 被研究。将定义涉及的特定PKC子类型。差异的原因 选择途径的激动剂将通过使用新颖的蛋白质 - 蛋白质相互作用来研究途径 蛋白酶测定系统。平行研究将使用原发性神经元培养物进行。封锁 在体内耐受性发育中表达DOR神经元中表达DOR神经元中的特定PKC亚型将是 探索。通过选择性地灭活吗啡信号通路，然后在信号后降低 含DOR神经元的事件，可能阻断吗啡耐受性而不改变吗啡 可以完成含有MOR的神经元中的活动。