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.

项目摘要 大麻素正在作为治疗慢性疼痛的阿片类药物的替代品。但是，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，我们会做 不完全了解大麻素和适应大麻素的跨脊索上部作用，大麻素在 炎。周围灰色（VLPAG）的腹外侧区域是脊柱上部的重要部位 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 divlpag中GABA释放的VLPAG 输出神经元导致镇痛（或动物模型中的抗伤害感受）。在AIM 1中，我将调查激动剂 - VLPAG中CB1R的诱导调节，包括脱敏，内在化以及对 蛋白质翻译。 AIM 2将研究持续炎症引起的CB1R信号传导的适应。我 假设CB1RS受蛋白质翻译的调节，蛋白质翻译受到持续炎症的失调。 突触前CB1R调节的这些特征将使CB1R与突触前MU- 阿片受体，不会脱敏。该提案的完成将阐明 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 实验将揭示持续性炎症引起的适应，这对 针对大麻素系统治疗疼痛的新治疗剂的开发。 为了研究CB1R调节和炎症引起的适应，我将学习全细胞斑块 夹紧电生理学，放射性物质结合，生物素化和紫霉素分析合并后 Susan Ingram博士和Marina Wolf博士的指导。以及合作和最高的研究 由俄勒冈健康与科学大学培养的环境和神经科学研究生计划 通过Vollum Institute博士。英格拉姆和沃尔夫将提供指导，使我成为一个 成功的独立科学家。 Vollum Institute举办了各种研讨会和学生举办的午餐 我将参加有关正在进行的研究并结识潜在的博士后顾问和未来的信息 合作者。我将通过Vollum-In-In-Orogress培训生研讨会来传播我的研究结果， 科学会议和书面出版物的演讲。最终，研究环境 俄勒冈州健康与科学大学以及博士的指导。英格拉姆和沃尔夫为 我可以发展成为一位成功的研究科学家，并成为以研究为中心的教授 学术机构。