Regulation of presynaptic cannabinoid CB1 receptors

项目摘要 大麻素正在成为阿片类药物治疗慢性疼痛的替代品。然而，许多 支持大麻素作为一种治疗选择的证据是轶事，在分子水平上， 不完全了解大麻素的超脊髓作用和大麻素在治疗过程中的适应性， 炎症中脑导水管周围灰质腹外侧区（vlPAG）是脊髓上神经节的重要部位， cannabinoids的作用 大麻素1受体（CB 1 R）位于vlPAG内的突触前末梢上，并且当 激活，抑制GABA从终端释放。抑制vlPAG中的GABA释放解除vlPAG的抑制 输出神经元导致镇痛（或动物模型中的抗伤害感受）。在目标1中，我将研究激动剂- VIPAG中CB 1 R的诱导调节，包括脱敏、内化，以及VIPAG中CB 1 R的作用。 蛋白质翻译目的2将研究持续性炎症诱导的CB 1 R信号转导适应。我 假设CB 1 R受蛋白质翻译调节，而蛋白质翻译因持续性炎症而失调。 突触前CB 1 R调节的这些特征将CB 1 R与突触前μ-受体区分开来。 阿片受体，它不会脱敏。完成这项建议将有助于了解 CB 1 R调节，中枢神经系统中最丰富的G蛋白偶联受体。此外，这些 实验将揭示持续性炎症引起的适应性变化，这对人类的免疫系统可能很重要。 开发靶向大麻素系统用于治疗疼痛的新疗法。 为了研究CB 1 R调节和炎症诱导的适应，我将学习全细胞贴片 钳位电生理学、放射性配体结合、生物素化和嘌呤霉素试验 苏珊·英格拉姆博士和玛丽娜·沃尔夫博士的指导。与合作和顶级研究一起， 由俄勒冈州健康与科学大学和神经科学研究生课程培养的环境 通过Vollum研究所，英格拉姆和沃尔夫博士将提供指导，使我成为一个 成功的独立科学家Vollum Institute举办各种研讨会和学生午餐会 我将参加，以了解正在进行的研究，并满足潜在的博士后顾问和未来 合作者我将通过Vollum正在进行的实习生研讨会传播我的研究成果， 在科学会议和书面出版物上发表演讲。最终，研究环境在 俄勒冈州健康与科学大学和英格拉姆和沃尔夫博士的指导提供了基础， 我可以发展成为一名成功的研究科学家，成为一名教授， 学术机构。

Project Summary Cannabinoids are emerging as alternatives to opioids for the treatment of chronic pain. However, much of the evidence supporting cannabinoids as a treatment option is anecdotal and, on the molecular level, we do not fully understand the supraspinal actions of cannabinoids and adaptations that cannabinoids undergo during inflammation. The ventrolateral region of the periaqueductal gray (vlPAG) is an important site for supraspinal actions of cannabinoids. Cannabinoid 1 receptors (CB1Rs) are located on presynaptic terminals within the vlPAG and, when activated, suppress GABA release from the terminal. Inhibition of GABA release in the vlPAG disinhibits vlPAG output neurons resulting in analgesia (or anti-nociception in animal models). In Aim 1, I will investigate agonist- induced regulation of the CB1R in the vlPAG including desensitization, internalization, as well as a role for protein translation. Aim 2 will investigate adaptations to CB1R signaling induced by persistent inflammation. I hypothesize that CB1Rs are regulated by protein translation which is dysregulated by persistent inflammation. These features of presynaptic CB1R regulation would distinguish the CB1R from the presynaptic mu- opioid receptor, which does not desensitize. Completion of this proposal will shed light onto mechanisms of the CB1R regulation, the most abundant G-protein coupled receptor in the central nervous system. Further, these experiments will reveal adaptations induced by persistent inflammation that could be important for the development of new therapeutics targeting the cannabinoid system for the treatment of pain. To investigate CB1R regulation and inflammation-induced adaptations, I will learn whole-cell patch clamp electrophysiology, radioligand binding, biotinylation, and puromycin assays under the combined mentorship of Dr. Susan Ingram and Dr. Marina Wolf. Together with the collaborative and top-rated research environment fostered by Oregon Health & Science University and the Neuroscience Graduate Program through the Vollum Institute, Drs. Ingram and Wolf will provide the guidance to empower me to become a successful independent scientist. The Vollum Institute hosts a variety of seminars and student-run lunches which I will attend to learn about ongoing research and meet potential post-doctoral advisors and future collaborators. I will disseminate my research findings through the Vollum Work-in-Progress trainee seminar, presentations at scientific conferences and written publications. Ultimately, the research environment at Oregon Health & Science University and the mentorship of Drs. Ingram and Wolf provides the foundation upon which I can develop into a successful research scientist and become a professor at a research-focused academic institution.