STED microscopy reveals mitotic stage-dependent CENP-A organization
- 1University of Groningen
Abstract
Chromosome segregation is vital. Its disruption can cause aneuploidy, a hallmark of cancer. Centromere protein A (CENP-A) is an important protein during the segregation as it marks the location of the centromere, where the kinetochore assembles for microtubule attachment. Each mammalian centromere contains hundreds of CENP-A nucleosomes, whose spatial arrangement is expected to be critical for kinetochore function and error-free chromosome segregation. However, previous studies--mostly in fixed cells and focused on metaphase--have yielded conflicting results on CENP-A organization. Given the centromere's sub-diffraction size, we used stimulated emission depletion (STED) super-resolution microscopy to visualize the CENP-A organization in both living (about 45 nm resolution) and fixed (about 30 nm resolution) immortalized human retinal pigment epithelial cells (hTERT RPE-1). We found that CENP-A organization does not adopt a single architecture but spans a spectrum from dense clusters to fragmented subclusters, with mitotic stage-dependent abundance and morphology. CENP-A chromatin is most dispersed in prophase, and progressively compacts during prometaphase and metaphase as microtubules attach. In meta- and anaphase predominantly dense organizations are formed, often with a plate-like morphology. Yet, non-dense and non-plate-like organizations persist through metaphase and anaphase, suggesting CENP-A spatial reorganization is heterogeneous during cell division.
Related
- A quantitative imaging framework reveals density-dependent GPCR oligomerization and organization in living cells
- 4D confocal microscopy of time dependent cell and nuclear osmotic response reveals differential osmoregulation and transport kinetics
- Mitotic MAX bookmarking drives MYC-dependent hypertranscription at TBP-bound promoters
- Replication-coupled chromatin assembly cooperates with ATR/CHK1 signaling to suppress DNA damage and premature MMB-dependent mitotic transcription
- Quantitative FLIM-FRET Microscopy to Monitor Nanoscale Chromatin Compaction In Vivo Reveals Structural Roles of Condensin Complexes
BibTeX
@misc{burgers2026sted,
title = {{STED microscopy reveals mitotic stage-dependent CENP-A organization}},
author = {T. C. Q. Burgers and S. J. Dijt and L. Vitrano and G. van den Bogaart and R. Vlijm},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.03.748884},
url = {https://www.biorxiv.org/content/10.64898/2026.09.03.748884v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - STED microscopy reveals mitotic stage-dependent CENP-A organization
AU - T. C. Q. Burgers
AU - S. J. Dijt
AU - L. Vitrano
AU - G. van den Bogaart
AU - R. Vlijm
PY - 2026
DA - 2026/09/03
PB - bioRxiv
DO - 10.64898/2026.09.03.748884
UR - https://www.biorxiv.org/content/10.64898/2026.09.03.748884v1
AB - Chromosome segregation is vital. Its disruption can cause aneuploidy, a hallmark of cancer. Centromere protein A (CENP-A) is an important protein during the segregation as it marks the location of the centromere, where the kinetochore assembles for microtubule attachment. Each mammalian centromere contains hundreds of CENP-A nucleosomes, whose spatial arrangement is expected to be critical for kinetochore function and error-free chromosome segregation. However, previous studies--mostly in fixed cells and focused on metaphase--have yielded conflicting results on CENP-A organization. Given the centromere's sub-diffraction size, we used stimulated emission depletion (STED) super-resolution microscopy to visualize the CENP-A organization in both living (about 45 nm resolution) and fixed (about 30 nm resolution) immortalized human retinal pigment epithelial cells (hTERT RPE-1). We found that CENP-A organization does not adopt a single architecture but spans a spectrum from dense clusters to fragmented subclusters, with mitotic stage-dependent abundance and morphology. CENP-A chromatin is most dispersed in prophase, and progressively compacts during prometaphase and metaphase as microtubules attach. In meta- and anaphase predominantly dense organizations are formed, often with a plate-like morphology. Yet, non-dense and non-plate-like organizations persist through metaphase and anaphase, suggesting CENP-A spatial reorganization is heterogeneous during cell division.
ER -