Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN 37232, USA.
Department of Medicine, Division of Infectious Diseases, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
STAR Protoc. 2021 Dec 10;2(4):100998. doi: 10.1016/j.xpro.2021.100998. eCollection 2021 Dec 17.
A key facet of epithelial differentiation is the assembly of actin-based protrusions known as microvilli, which amplify apical membrane surface area for various cell functions. To probe mechanisms of microvillus assembly, we developed a protocol using spinning disk confocal microscopy to directly visualize microvillus biogenesis on the surface of cultured porcine kidney epithelial cell monolayers engineered to express fluorescent proteins. This protocol offers access to the molecular details of individual protrusion growth events at high spatiotemporal resolution. For complete details on the use and execution of this protocol, please refer to Gaeta et al. (2021).
上皮细胞分化的一个关键方面是肌动蛋白为基础的突起的组装，这些突起被称为微绒毛，它们为各种细胞功能扩大了顶端膜表面积。为了探究微绒毛组装的机制，我们开发了一种使用旋转盘共聚焦显微镜的方案，该方案可直接在表达荧光蛋白的培养猪肾上皮细胞单层表面上可视化微绒毛的生物发生。该方案可在高时空分辨率下获得单个突起生长事件的分子细节。有关该方案的使用和执行的完整详细信息，请参阅 Gaeta 等人（2021 年）。
