Regulation of presynaptic cannabinoid CB1 receptors

突触前大麻素 CB1 受体的调节

• 批准号: 10065422
• 负责人: Courtney Anne Bouchet
• 金额: $ 4.55万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10065422
• 关键词: Absence of pain sensation; Acute; Address; Agonist; Animal Model; Binding; Biological Assay; Biotinylation; Brain region; CNR1 gene; Cannabinoids; Cell Line; Cells; Dangerousness; Development; Educational process of instructing; Electrophysiology (science); Environment; Exposure to; FRAP1 gene; Fostering; Foundations; Freund' s Adjuvant; Funding; Future; G-Protein-Coupled Receptors; Goals; Health Sciences; Inflammation; Institutes; Institution; Kinetics; Knowledge; Learning; Light; Link; Mediating; Membrane; Mentors; Mentorship; Molecular; Neuraxis; Neurons; Neurosciences; Nociception; Opioid; Optics; Oregon; Output; Pain; Pain management; Postdoctoral Fellow; Postsynaptic Membrane; Presynaptic Receptors; Presynaptic Terminals; Process; Proteins; Publications; Puromycin; Receptor Inhibition; Receptor Signaling; Regulation; Research; Role; Running; Scientist; Signal Transduction; Sirolimus; Site; Students; Surface; Synapses; System; Time; Training; Translations; Universities; Wolves; Work; career; chronic pain; desensitization; experimental study; gamma-Aminobutyric Acid; inhibitor/antagonist; midbrain central gray substance; mu opioid receptors; new therapeutic target; patch clamp; presynaptic; professor; programs; radioligand; receptor binding; receptor function; receptor internalization; receptor recycling; response; skills; symposium; translation assay

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.

项目概要 大麻素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轶事，在分子水平上，我们确实 不完全了解大麻素的脊髓上作用以及大麻素在过程中经历的适应 炎。导水管周围灰质的腹外侧区 (vlPAG) 是脊髓上皮炎的重要部位。 大麻素的作用。 大麻素 1 受体 (CB1R) 位于 vlPAG 内的突触前末端，当 激活，抑制 GABA 从终端释放。抑制 vlPAG 中 GABA 的释放会抑制 vlPAG 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- vlPAG 中 CB1R 的诱导调节，包括脱敏、内化以及 蛋白质翻译。目标 2 将研究持续炎症诱导的对 CB1R 信号传导的适应。我 假设 CB1R 受到蛋白质翻译的调节，而蛋白质翻译则因持续炎症而失调。 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。该提案的完成将揭示该机制的机制 CB1R 调节，中枢神经系统中最丰富的 G 蛋白偶联受体。此外，这些 实验将揭示持续炎症引起的适应，这对于 开发针对大麻素系统的新疗法来治疗疼痛。 为了研究 CB1R 调节和炎症诱导的适应，我将学习全细胞补丁 结合钳电生理学、放射性配体结合、生物素化和嘌呤霉素测定 苏珊·英格拉姆博士和玛丽娜·沃尔夫博士的指导。连同合作和顶级研究 俄勒冈健康与科学大学和神经科学研究生项目营造的环境 通过沃勒姆研究所，博士。英格拉姆和沃尔夫将提供指导，使我能够成为一名 成功的独立科学家。沃勒姆研究所举办各种研讨会和学生举办的午餐会 我将参加该活动以了解正在进行的研究并会见潜在的博士后顾问和未来 合作者。我将通过 Vollum Work-in-Progress 实习生研讨会传播我的研究成果， 在科学会议上的演讲和书面出版物。最终，研究环境 俄勒冈健康与科学大学和博士的指导。英格拉姆和沃尔夫奠定了基础 我可以发展成为一名成功的研究科学家，并成为专注于研究的教授 学术机构。