Department of Neurology and Program in Neuroscience, University of California, San Francisco, San Francisco, California, USA.
Department of Pediatrics, University of California, San Francisco, San Francisco, California, USA.
Nat Med. 2014 Aug;20(8):954-960. doi: 10.1038/nm.3618. Epub 2014 Jul 6.
Functional screening for compounds that promote remyelination represents a major hurdle in the development of rational therapeutics for multiple sclerosis. Screening for remyelination is problematic, as myelination requires the presence of axons. Standard methods do not resolve cell-autonomous effects and are not suited for high-throughput formats. Here we describe a binary indicant for myelination using micropillar arrays (BIMA). Engineered with conical dimensions, micropillars permit resolution of the extent and length of membrane wrapping from a single two-dimensional image. Confocal imaging acquired from the base to the tip of the pillars allows for detection of concentric wrapping observed as 'rings' of myelin. The platform is formatted in 96-well plates, amenable to semiautomated random acquisition and automated detection and quantification. Upon screening 1,000 bioactive molecules, we identified a cluster of antimuscarinic compounds that enhance oligodendrocyte differentiation and remyelination. Our findings demonstrate a new high-throughput screening platform for potential regenerative therapeutics in multiple sclerosis.
功能筛选促进髓鞘再生的化合物是多发性硬化症合理治疗开发的主要障碍。髓鞘再生的筛选存在问题，因为髓鞘形成需要轴突的存在。标准方法无法解决细胞自主性效应，不适合高通量格式。在这里，我们使用微柱阵列（BIMA）描述了一种用于髓鞘形成的二进制指示物。通过工程设计成锥形尺寸，微柱允许从单个二维图像中解析膜包裹的程度和长度。从柱子的底部到顶部获取的共聚焦成像允许检测到观察到的同心包裹，表现为“环”状髓鞘。该平台以 96 孔板的形式格式化，适用于半自动随机采集和自动检测和定量。在筛选 1000 种生物活性分子后，我们确定了一组抗毒蕈碱化合物，这些化合物可增强少突胶质细胞分化和髓鞘再生。我们的发现展示了一种新的高通量筛选平台，可用于多发性硬化症的潜在再生治疗。
