Super-resolution microscopy reveals nanoscale architecture and regulation of podosome clusters in primary macrophages

Fen Hu; Donglan Zhu; Hao Dong; Ping Zhang; Fulin Xing; Wan Li; Rui Yan; Jun Zhou; Ke Xu; Leiting Pan; Jingjun Xu

doi:10.1016/j.isci.2022.105514

Super-resolution microscopy reveals nanoscale architecture and regulation of podosome clusters in primary macrophages

iScience. 2022 Nov 5;25(12):105514. doi: 10.1016/j.isci.2022.105514. eCollection 2022 Dec 22.

Authors

Fen Hu¹, Donglan Zhu¹, Hao Dong¹, Ping Zhang¹, Fulin Xing¹, Wan Li², Rui Yan², Jun Zhou³, Ke Xu², Leiting Pan^{1

3

4}, Jingjun Xu^{1

4}

Affiliations

¹ The Key Laboratory of Weak-Light Nonlinear Photonics of Education Ministry, School of Physics and TEDA Institute of Applied Physics, Nankai University, Tianjin 300071, China.
² Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.
³ State Key Laboratory of Medicinal Chemical Biology, Frontiers Science Center for Cell Responses, College of Life Sciences, Nankai University, Tianjin 300071, China.
⁴ Shenzhen Research Institute of Nankai University, Shenzhen, Guangdong 518083, China.

Abstract

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.

Keywords: Biological sciences; Cell biology; Immunology; Organizational aspects of cell biology.