Super-resolution microscopy reveals nanoscale architecture and regulation of podosome clusters in primary macrophages.
Super-resolution microscopy reveals nanoscale architecture and regulation of podosome clusters in primary macrophages.
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超分辨率显微镜揭示了初级巨噬细胞中足体簇的纳米级结构和调控
DOI：
10.1016/j.isci.2022.105514
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发表时间：
2022-12-22
期刊：
影响因子：
5.8
通讯作者：
Xu, Jingju
中科院分区：
文献类型：
--
作者：
Hu, Fen;Zhu, Donglan;Dong, Hao;Zhang, Ping;Xing, Fulin;Li, Wan;Yan, Rui;Zhou, Jun;Xu, Ke;Pan, Leiting;Xu, Jingju
足体是一种重要的肌动蛋白黏附结构，在细胞迁移和基质侵袭中发挥着重要作用。在这里，我们阐明了原代巨噬细胞足体簇的超微结构组织和调控。利用三维随机光学重建显微镜(3D-STORM)，我们获得了∼20/50 nm(横向/轴向)的空间分辨率，以解决足体核心和环组分之间的相互定位，并进一步表明微管在肌球蛋白IIA层穿过足体。微管断裂导致的足小体溶解以前被归因于Rho/Rock-myosin信号转导，但用Y27632或Blebbistatin抑制这一途径只能部分恢复足小体的组装，因此提示微管的物理支持在稳定足小体结构中起到了作用。通过改进的底物包被技术，我们进一步证实了巨噬细胞对基质的降解能力依赖于足体簇的形成。总之，3D-STORM超分辨率显微镜揭示了巨噬细胞中降解基质的足体簇的纳米尺度空间排列和微管依赖的调节。3D-STORM描绘了一幅雪人般的巨噬细胞足体核心3D-STORM图，足体环组件的纳米级排列，微管在肌球蛋白层穿过足体并使其稳定，巨噬细胞基质的降解取决于足体簇的形成生物科学；免疫学；细胞生物学；细胞生物学的组织方面
Podosomes, an important actin-based adhesive architecture, play critical roles in cell migration and matrix invasiveness. Here, we elucidate the ultrastructural organization and regulation of podosome clusters in primary macrophages. With three-dimensional stochastic optical reconstruction microscopy (3D-STORM), we achieve ∼20/50 nm (lateral/axial) spatial resolution to resolve the mutual localization of podosome core and ring components, and further show that microtubules pass through podosomes at the layer of myosin IIA. The microtubule disruption-caused podosome dissolution is previously ascribed to Rho/ROCK-myosin signaling, yet inhibiting this pathway with Y27632 or blebbistatin only partially recovers podosome assembly, thus suggesting the contribution of the physical supporting of microtubules in stabilizing podosome structures. Through improved substrate-coating technique, we further corroborate that the matrix-degrading capability of macrophages depends on the formation of podosome clusters. Together, 3D-STORM super-resolution microscopy reveals the nanoscale spatial arrangement and the microtubule-dependent regulation of the matrix-degrading podosome clusters in macrophages. 3D-STORM delineates a snowman-like portrait of macrophage podosome core 3D-STORM maps the nanoscale arrangement of podosome ring components Microtubules pass through podosomes at the layer of myosin and stabilize them Macrophage matrix degradation depends on the formation of podosome clusters Biological sciences; Immunology; Cell biology; Organizational aspects of cell biology
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影响因子：
9.2
作者：
Collin, Olivier;Na, Sungsoo;Chowdhury, Farhan;Hong, Michael;Shin, Myung Eun;Wang, Fei;Wang, Ning
通讯作者：
Wang, Ning
影响因子：
5.3
作者：
Duplan, Martin Biosse;Zalli, Detina;Baron, Roland
通讯作者：
Baron, Roland
影响因子：
17.1
作者：
Bouissou, Angs;Proag, Amsha;Poincloux, Renaud
通讯作者：
Poincloux, Renaud
影响因子：
5.3
作者：
Daubon, Thomas;Buccione, Roberto;Genot, Elisabeth
通讯作者：
Genot, Elisabeth
影响因子：
6.6
作者：
Evans, JG;Matsudaira, P
通讯作者：
Matsudaira, P