Quantifying cell-cycle-dependent chromatin dynamics during interphase by live 3D tracking

• Published in: iScience 2022-05, Vol.25 (5), p.104197-104197, Article 104197
• Main Authors: Naor, Tal, Nogin, Yevgeni, Nehme, Elias, Ferdman, Boris, Weiss, Lucien E., Alalouf, Onit, Shechtman, Yoav
• Format: Article
• Language: English
• Subjects: Biological sciences; Biophysics; Chromosome organization; Optical imaging
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2022.104197

The study of cell cycle progression and regulation is important to our understanding of fundamental biophysics, aging, and disease mechanisms. Local chromatin movements are generally considered to be constrained and relatively consistent during all interphase stages, although recent advances in our understanding of genome organization challenge this claim. Here, we use high spatiotemporal resolution, 4D (x, y, z and time) localization microscopy by point-spread-function (PSF) engineering and deep learning-based image analysis, for live imaging of mouse embryonic fibroblast (MEF 3T3) and MEF 3T3 double Lamin A Knockout (LmnaKO) cell lines, to characterize telomere diffusion during the interphase. We detected varying constraint levels imposed on chromatin, which are prominently decreased during G0/G1. Our 4D measurements of telomere diffusion offer an effective method to investigate chromatin dynamics and reveal cell-cycle-dependent motion constraints, which may be caused by various cellular processes. [Display omitted] •PSF engineering allows scan-free, high spatiotemporal live 3D telomere tracking•During the G0/G1 phase, telomere motion is less constrained than in other phases•There is observable difference between lateral (xy) and axial (z) chromatin motion•In Lamin A-depleted cells, motion constraint was reduced Optical imaging; Biological sciences; Chromosome organization; Biophysics