New cell biology tools to study myelin development, dynamics, and disease

项目摘要/摘要 髓磷脂--神经元轴突周围的电绝缘体--对于脊椎动物来说是快速神经信号传递所必需的， 在多发性硬化症和受伤后的疾病中，它的丢失会导致患者严重残疾。髓磷脂 作为被动电绝缘体的传统角色最近被重新想象为一个动态过程，在这个过程中 少突胶质细胞响应神经元活动建立和重塑髓鞘，并在代谢上提供支持 它们形成髓鞘的神经元。这些新发现开启了许多令人兴奋的新研究问题，例如： 少突胶质细胞如何调节其形态以调节传导速度或其他神经元功能， 在发育过程中还是在成人中枢神经系统(CNS)中？髓鞘动力学在其中扮演了什么角色 除了多发性硬化症以外的高级大脑功能和神经退行性疾病？根本性的差距 目前的知识在于了解少突胶质细胞生物学是如何在所有环境中调节的 发展、动态和疾病。弥合这一知识鸿沟需要构建创新的新工具来 突破了阻碍我们研究少突胶质细胞生物学能力的实验限制 到目前为止，VIVO。这项提议的目标是创建一种用于研究少突胶质细胞生物学的病毒(AAV)工具包 在活体内。该工具包将允许对单个少突胶质细胞进行明亮、稀疏的标记和操作 小鼠中枢神经系统，这将有助于研究确定控制少突胶质细胞形态的机制 (目标1)。此外，这个工具包将允许对记者或微扰者进行亚细胞定位，以在功能上区分开来 少突胶质细胞内的区域，包括髓鞘(目标2)。我们认为这个病毒细胞生物学工具包 将在髓鞘和更广泛的神经科学领域产生广泛的积极影响：i)快速加速 与传统的小鼠遗传学相比，从想法到发现的时间，ii)通过以下方式简化实验设计 即插即用的模块化结构体系结构，iii)解锁可视化和/或干扰单个鼠标的能力 体内的少突胶质细胞，以及iv)创造新的工具来研究不同功能的亚细胞结构 髓鞘。通过这项研究，我们将创建和优化一个简单但功能强大的工具包来研究髓鞘细胞 在小鼠中枢神经系统的活体生物学。我们希望这些工具能够迅速被现场采用，并开放 髓鞘研究中令人兴奋的新领域。我们将把这些工具作为免费资源共享，从而最大限度地 它们的潜在影响揭示了对髓鞘发育、动力学和疾病的新的细胞生物学见解。

PROJECT SUMMARY/ABSTRACT Myelin—the electrical insulator around neuronal axons—is essential in vertebrates for rapid nerve signaling, and its loss in diseases like multiple sclerosis and following injury causes severe disability in patients. Myelin’s traditional role as a passive electrical insulator has recently been reimagined as a dynamic process, in which oligodendrocytes build and remodel myelin sheaths in response to neuronal activity, and metabolically support the neurons they myelinate. These new discoveries open up many exciting new research questions, such as: how do oligodendrocytes regulate their morphology to tune conduction velocity or other neuronal functions, during development or in the adult central nervous system (CNS)? What roles do myelin dynamics play in higher-order brain functions and in neurodegenerative diseases beyond multiple sclerosis? A fundamental gap in current knowledge lies in understanding how oligodendrocyte cell biology is regulated in all of the contexts of development, dynamics, and disease. Bridging this knowledge gap requires building innovative new tools to break through the experimental limitations that have hindered our ability to study oligodendrocyte cell biology in vivo thus far. The goal of this proposal is to create a viral (AAV) toolkit for studying oligodendrocyte cell biology in vivo. This toolkit will allow for bright, sparse labeling and manipulation of single oligodendrocytes in the mouse CNS, which will empower studies to determine the mechanisms controlling oligodendrocyte morphology (Aim 1). In addition, this toolkit will allow subcellular targeting of reporters or perturbants to functionally-distinct regions within oligodendrocytes, including myelin sheaths (Aim 2). We propose that this viral cell biology toolkit will have a broad positive impact across the myelin and broader neuroscience fields, by: i) rapidly accelerating the time from idea to discovery, compared to traditional mouse genetics, ii) simplifying experimental design via “plug-and-play” modular construct architecture, iii) unlocking the ability to visualize and/or perturb single mouse oligodendrocytes in vivo, and iv) creating new tools to study functionally distinct subcellular structures in myelin. From this research, we will create and optimize a simple but powerful toolkit for studying myelin cell biology in vivo in the mouse CNS. It is our hope that these tools can be rapidly adopted by the field, and open exciting new frontiers in myelin research. We will share these tools as a free resource, thereby maximizing their potential impact to reveal new cell biological insights into myelin development, dynamics, and disease.