Reiser Jochen, Lee Ha Won, Gupta Vineet, Altintas Mehmet M
Department of Medicine, Rush University Medical Center, Chicago, IL.
Department of Medicine, Rush University Medical Center, Chicago, IL.
Adv Chronic Kidney Dis. 2017 May;24(3):183-188. doi: 10.1053/j.ackd.2017.04.001.
Podocytes form the visceral layer of a kidney glomerulus and express a characteristic octopus-like cellular architecture specialized for the ultrafiltration of blood. The cytoskeletal dynamics and structural elasticity of podocytes rely on the self-organization of highly interconnected actin bundles, and the maintenance of these features is important for the intact glomerular filtration. Development of more differentiated podocytes in culture has dramatically increased our understanding of the molecular mechanisms regulating podocyte actin dynamics. Podocytes are damaged in a variety of kidney diseases, and therapies targeting podocytes are being investigated with increasing efforts. Association between podocyte damage and disease severity-or between podocyte recovery and the performance of therapeutic molecules-have been the venues of research for years. In this perspective, more standardized high--content screening has emerged as a powerful tool for visualization and analysis of podocyte morphology. This high-throughput fluorescence microscopy technique is based on an automated image analysis with simultaneous detection of various phenotypes (multiplexing) across multiple phenotypic parameters (multiparametric). Here, we review the principles of high-content screening technology and summarize efforts to carry out small compound screen using podocytes.
足细胞构成肾小球的脏层,呈现出一种特殊的章鱼状细胞结构,专门用于血液超滤。足细胞的细胞骨架动力学和结构弹性依赖于高度相互连接的肌动蛋白束的自组织,维持这些特征对于完整的肾小球滤过很重要。在培养中生成更分化的足细胞极大地增进了我们对调节足细胞肌动蛋白动力学分子机制的理解。足细胞在多种肾脏疾病中受损,针对足细胞的治疗方法正在加大研究力度。足细胞损伤与疾病严重程度之间的关联,或者足细胞恢复与治疗分子疗效之间的关联,多年来一直是研究的重点。从这个角度来看,更标准化的高内涵筛选已成为可视化和分析足细胞形态的强大工具。这种高通量荧光显微镜技术基于自动图像分析,可同时检测多个表型参数(多参数)上的各种表型(多重检测)。在此,我们回顾高内涵筛选技术的原理,并总结利用足细胞进行小分子化合物筛选的工作。