Investigating the role of cannabinoid receptors in oligodendrocyte development

研究大麻素受体在少突胶质细胞发育中的作用

• 批准号: 10605105
• 负责人: Tania G Miramontes
• 金额: $ 4.77万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605105
• 关键词: 2-arachidonylglycerol; Affect; Agonist; Amides; Animal Model; Axon; Binding; CNR1 gene; CNR2 gene; CRISPR/Cas technology; Calcium; Cannabinoids; Cell Lineage; Cell physiology; Cells; Cellular Membrane; Cellular biology; Central Nervous System; Demyelinating Diseases; Demyelinations; Development; Dimensions; Disease; Dorsal; Endocannabinoids; Enzymes; Event; Experimental Genetics; Fatty Acids; Foundations; Future; G protein-coupled inwardly-rectifying potassium channel; G-Protein-Coupled Receptors; GPR55 receptor; Genetic; Genomics; Health; Image; Impairment; Larva; Ligands; Light; Lipids; Mediating; Membrane; Metabolic; Methods; Modeling; Molecular; Monoacylglycerol Lipases; Multiple Sclerosis; Myelin; Myelin Basic Proteins; NAPE-PLD; Nervous System; Neuroglia; Neurologic Symptoms; Neuronal Dysfunction; Neurons; Oligodendroglia; Optics; Peripheral Nervous System; Peripheral Nervous System Diseases; Physiological; Proteins; Proteomics; Rattus; Receptor Activation; Receptor Signaling; Resolution; Role; Schwann Cells; Series; Signal Transduction; Spinal Cord; Symptoms; System; Testing; Therapeutic; Time; Visualization; Zebrafish; anandamide; axonal degeneration; cannabinoid receptor; cell motility; covalent bond; endogenous cannabinoid system; fatty acid amide hydrolase; genetic analysis; genetic approach; genome editing; glial cell development; in vivo; innovation; interest; lipoprotein lipase; loss of function; mutant; myelination; nervous system disorder; neuroregulation; novel; oligodendrocyte lineage; oligodendrocyte myelination; oligodendrocyte precursor; palmidrol; pharmacologic; precursor cell; prevent; protein expression; receptor; receptor expression; reduce symptoms; remyelination; small molecule; tool; tool development

Project Summary Myelin is the protective sheath that wraps around neuronal axons to facilitate fast, saltatory conduction, maintain axon integrity, and provide metabolic support. It is made by specialized glial cells called oligodendrocytes (OL) in the central nervous system (CNS). The importance of myelin is highlighted in neurological diseases such as multiple sclerosis (MS), where disruption of myelin can result in improper neuronal function and may ultimately lead to axonal degeneration along with other symptoms. While the importance of myelin is thus evident, there are currently no treatment options to prevent or aid in remyelination, in part because the molecular mechanisms of myelin formation are not fully understood. However, the endocannabinoid system (ECS) has gained increasing interest as a potential target to treat several neurological diseases, including MS. Yet there is no clear understanding in the relationship between the ECS and OL development or myelination. Aiming to understand of the molecular mechanisms that drive OL development and myelination, our lab conducted a proteomic analysis of cultured rat primary OLs in which myelination was increased by genetic methods. Among the list of proteins that were significantly upregulated in OLs with increased myelin were two ECS enzymes, FAAH1 and DAGL-. This increase occurred during OL precursor (OPC) differentiation, suggesting the ECS may play a role at this stage of OL development. This model is supported by my preliminary in vivo studies in zebrafish activating the ECS. For this proposal, I aim to define the role of CB1 and CB2 in OL development and myelination through continued pharmacological receptor manipulation, careful genomic analysis using CRISPR-Cas9 mediated genome editing, and precise multidimensional receptor activation and live imaging. In Aim 1, I will use a range of pharmacological and genetic approaches to dissect the roles of CB1 and CB2 in various stages of OL development and myelination. In Aim 2, I will investigate the downstream signaling effects of CB1 activation through calcium imaging, using a novel photocaged ligand that is genetically targeted to OL lineage cells.

项目概要 髓磷脂是包裹神经元轴突的保护鞘，以促进快速、跳跃式传导， 维持轴突完整性，并提供代谢支持。它由称为“特殊神经胶质细胞”的细胞制成。 中枢神经系统 (CNS) 中的少突胶质细胞 (OL) 强调了髓磷脂的重要性。 神经系统疾病，如多发性硬化症 (MS)，髓鞘质破坏可能导致不适当的治疗 神经元功能，最终可能导致轴突变性以及其他症状。 因此，髓磷脂的重要性是显而易见的，目前没有治疗方案可以预防或帮助髓鞘再生， 部分原因是髓磷脂形成的分子机制尚未完全了解。 内源性大麻素系统（ECS）作为治疗多种疾病的潜在靶点引起了越来越多的关注。 包括 MS 在内的神经系统疾病与 ECS 之间的关系尚无明确认识。 OL 发育或髓鞘形成的目的是了解驱动 OL 的分子机制。 发育和髓鞘形成，我们的实验室对培养的大鼠原代 OL 进行了蛋白质组学分析，其中 通过遗传方法增加髓鞘形成是显着上调的蛋白质列表之一。 髓磷脂增加的 OL 是两种 ECS 酶，FAAH1 和 DAGL-，这种增加发生在 OL 期间。 前体（OPC）分化，表明 ECS 可能在 OL 发育的这个阶段发挥作用。 该模型得到了我对斑马鱼激活 ECS 的初步体内研究的支持。 通过持续的药理学定义 CB1 和 CB2 在 OL 发育和髓鞘形成中的作用 受体操作、使用 CRISPR-Cas9 介导的基因组编辑进行仔细的基因组分析以及精确的 在目标 1 中，我将使用一系列药理和实时成像。 遗传学方法剖析 CB1 和 CB2 在 OL 发育和髓鞘形成的各个阶段中的作用。 在目标 2 中，我将通过钙成像研究 CB1 激活的下游信号传导效应，使用 新型光笼配体，其基因靶向 OL 谱系细胞。