Curriculum in Bioinformatics and Computational Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Computational Medicine Program, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Nat Commun. 2022 Jul 27;13(1):4363. doi: 10.1038/s41467-022-32038-0.
Podosomes are actin-enriched adhesion structures important for multiple cellular processes, including migration, bone remodeling, and phagocytosis. Here, we characterize the structure and organization of phagocytic podosomes using interferometric photoactivated localization microscopy, a super-resolution microscopy technique capable of 15-20 nm resolution, together with structured illumination microscopy and localization-based super-resolution microscopy. Phagocytic podosomes are observed during frustrated phagocytosis, a model in which cells attempt to engulf micropatterned IgG antibodies. For circular patterns, this results in regular arrays of podosomes with well-defined geometry. Using persistent homology, we develop a pipeline for semi-automatic identification and measurement of podosome features. These studies reveal an hourglass shape of the podosome actin core, a protruding knob at the bottom of the core, and two actin networks extending from the core. Additionally, the distributions of paxillin, talin, myosin II, α-actinin, cortactin, and microtubules relative to actin are characterized.
足突是富含肌动蛋白的黏附结构，对于多种细胞过程很重要，包括迁移、骨重塑和吞噬作用。在这里，我们使用干涉光激活定位显微镜（一种能够达到 15-20nm 分辨率的超分辨率显微镜技术），结合结构光照明显微镜和基于定位的超分辨率显微镜，对吞噬足突的结构和组织进行了研究。吞噬足突在受挫性吞噬过程中被观察到，这是一种细胞试图吞噬微图案化 IgG 抗体的模型。对于圆形图案，这会导致具有明确定义几何形状的规则排列的足突。我们使用持久同调开发了一个用于半自动化识别和测量足突特征的流水线。这些研究揭示了足突肌动蛋白核心的沙漏形状、核心底部的突出旋钮，以及从核心延伸出的两个肌动蛋白网络。此外，还对相对于肌动蛋白的桩蛋白、黏着斑蛋白、肌球蛋白 II、α-辅肌动蛋白、纽蛋白和微管的分布进行了描述。
