Investigating the role of cannabinoid receptors in oligodendrocyte development

项目摘要 髓鞘是包裹在神经元轴突周围的保护鞘，以促进快速、跳跃的传导， 保持轴突的完整性，并提供代谢支持。它是由特殊的神经胶质细胞制造的，称为 中枢神经系统内的少突胶质细胞(OL)。髓鞘的重要性在 神经系统疾病，如多发性硬化症(MS)，髓鞘中断可导致 神经功能受损，最终可能导致轴突变性和其他症状。而当 因此髓鞘的重要性是显而易见的，目前还没有预防或帮助重新髓鞘形成的治疗选择， 部分原因是髓鞘形成的分子机制还不完全清楚。然而， 内源性大麻素系统(ECS)作为治疗多种疾病的潜在靶点而受到越来越多的关注。 包括MS在内的神经系统疾病，但对ECS之间的关系尚无明确了解 和OL发育或髓鞘形成。旨在了解推动OL的分子机制 我们实验室对培养的大鼠原代成骨细胞进行了蛋白质组学分析，其中 髓鞘形成是通过遗传方法增加的。在一系列显著上调的蛋白质中 髓鞘增多的OL是两种ECS酶：FAAH1和DAGL-。这一增长发生在OL期间 前体(OPC)分化，提示ECS可能在OL发育的这个阶段发挥作用。这 我在斑马鱼体内激活ECS的初步研究支持了这一模型。对于这项提议，我的目标是 通过持续的药理作用确定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，并精确 多维受体激活和实时成像。在目标1中，我将使用一系列的药理和 遗传学方法剖析CB1和CB2在OL发育和髓鞘形成的不同阶段中的作用。 在目标2中，我将通过钙成像研究CB1激活的下游信号效应，使用 以OL系细胞为基因靶点的新型光笼配基。

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.