Regulation of presynaptic cannabinoid CB1 receptors

项目摘要 大麻类药物正在成为治疗慢性疼痛的阿片类药物的替代品。然而，很多 支持大麻素作为治疗选择的证据是坊间传闻，在分子水平上，我们确实是这样做的。 不完全了解大麻素的脊椎上作用和大麻素在 发炎。中脑导水管周围灰腹外侧区(VlPAG)是脊髓上的一个重要部位。 大麻素的作用。 大麻素1受体(CB1Rs)位于vlPAG内的突触前终末，当 激活，抑制GABA从终端释放。抑制vlPAG中GABA的释放可解除vlPAG的抑制 产生镇痛(或动物模型中的抗伤害性)的输出神经元。在目标1中，我将调查激动剂- 对vlPAG中CB1R的诱导调节包括脱敏、内化以及 蛋白质翻译。目的2将研究持续性炎症诱导的CB1R信号的适应性。我 假设CB1Rs由蛋白质翻译调节，而蛋白质翻译受到持续炎症的失调。 突触前CB1R调节的这些特征将CB1R与突触前MU区分开来。 阿片受体，不会脱敏。这项提案的完成将有助于揭示 CB1R调节，中枢神经系统中含量最丰富的G蛋白偶联受体。此外，这些 实验将揭示持续炎症诱导的适应，这可能对 针对大麻素系统治疗疼痛的新疗法的开发。 为了研究CB1R的调节和炎症诱导的适应，我将学习全细胞贴片 钳夹电生理、放射性配基结合、生物素化和嘌呤霉素联合检测 苏珊·英格拉姆博士和玛丽娜·沃尔夫博士的指导。与合作和一流的研究 俄勒冈健康与科学大学和神经科学研究生项目培育的环境 通过Vollum研究所，英格拉姆博士和沃尔夫博士将提供指导，使我成为一名 成功的独立科学家。Vollum Institute举办各种研讨会和学生举办的午餐 我将参加其中，了解正在进行的研究，并会见潜在的博士后导师和未来 合作者。我将通过Vollum正在进行的实习生研讨会来传播我的研究成果， 在科学会议和书面出版物上发表演讲。归根结底， 俄勒冈健康与科学大学以及英格拉姆和沃尔夫博士的指导为 我可以发展成为一名成功的研究科学家，并成为一名专注于研究的教授 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.