Transport and Organization of Individual Vimentin Filaments Within Dense Networks Revealed by Single Particle Tracking and 3D FIB-SEM

Abstract

Vimentin intermediate filaments (VIFs) form complex, tight-packed networks; due to this density, traditional ensemble labeling and imaging approaches cannot accurately discern single filament behavior. To address this, we introduce a sparse vimentin-SunTag labeling strategy to unambiguously visualize individual filament dynamics. This technique confirmed known long-range dynein and kinesin transport of peripheral VIFs and uncovered extensive bidirectional VIF motion within the perinuclear vimentin network, a region we had thought too densely bundled to permit such motility. To examine the nanoscale organization of perinuclear vimentin, we acquired high-resolution electron microscopy volumes of a vitreously frozen cell and reconstructed VIFs and microtubules within a ∼50 µm3 window. Of 583 VIFs identified, most were integrated into long, semi-coherent bundles that fluctuated in width and filament packing density. Unexpectedly, VIFs displayed minimal local co-alignment with microtubules, save for sporadic cross-over sites that we predict facilitate cytoskeletal crosstalk. Overall, this work demonstrates single VIF dynamics and organization in the cellular milieu for the first time.

Citation

Bhuvanasundar Renganathan, Andrew Moore, Wei-Hong Yeo, Alyson Petruncio, David Ackerman, Aubrey Wiegel, The CellMap Team, H. Amalia Pasolli, C. Shan Xu, Gleb Shtengel, Harald F. Hess, Anna S Serpinskaya, Hao F. Zhang, Jennifer Lippincott-Schwartz, and Vladimir I. Gelfand. 2025. “Transport and Organization of Individual Vimentin Filaments Within Dense Networks Revealed by Single Particle Tracking and 3D FIB-SEM.” Journal of Cell Biology. https://doi.org/10.1083/jcb.202406054.

@article{Renganathan2025,
author = {Bhuvanasundar Renganathan and Andrew Moore and Wei-Hong Yeo and Alyson Petruncio and David Ackerman and Aubrey Wiegel and The CellMap Team and H. Amalia Pasolli and C. Shan Xu and Gleb Shtengel and Harald F. Hess and Anna S Serpinskaya and Hao F. Zhang and Jennifer Lippincott-Schwartz and Vladimir I. Gelfand},
doi = {10.1083/jcb.202406054},
journal = {Journal of Cell Biology},
title = {Transport and Organization of Individual Vimentin Filaments Within Dense Networks Revealed by Single Particle Tracking and 3D FIB-SEM},
year = {2025}}