Abstract
Chromosome segregation ensures the equal partitioning of chromosomes at mitosis. However, long chromosome arms may pose a problem for complete sister chromatid separation. In this paper we report on the analysis of cell division in primary cells from field vole Microtus agrestis, a species with 52 chromosomes including two giant sex chromosomes. Dual chromosome painting with probes specific for the X and the Y chromosomes showed that these long chromosomes are prone to mis-segregate, producing DNA bridges between daughter nuclei and micronuclei. Analysis of mitotic cells with incomplete chromatid separation showed that reassembly of the nuclear membrane, deposition of INner CENtromere Protein (INCENP)/Aurora B to the spindle midzone and furrow formation occur while the two groups of daughter chromosomes are still connected by sex chromosome arms. Late cytokinetic processes are not efficiently inhibited by the incomplete segregation as in a significant number of cell divisions cytoplasmic abscission proceeds while Aurora B is at the midbody. Live-cell imaging during late mitotic stages also revealed abnormal cell division with persistent sister chromatid connections. We conclude that late mitotic regulatory events do not monitor incomplete sister chromatid separation of the large X and Y chromosomes of Microtus agrestis, leading to defective segregation of these chromosomes. These findings suggest a limit in chromosome arm length for efficient chromosome transmission through mitosis.
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Acknowledgements
This work was performed at the Cambridge Resource Centre for Comparative Genomics and was supported by a grant from the Wellcome Trust to MAFS.
Microtus agrestis X- and Y-chromosome-specific DNA was kindly provided by Dr. F. Yang. The inner centromere protein (INCENP) antibody was kindly provided by Prof. WC Earnshaw. A Microtus agrestis specimen was supplied by the house cat Tipper.
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Communicated by F. Uhlmann
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Supplementary movie 1, 2, and 3 show Microtus agrestis cell division. The chromosomes were visualized using HistoneH2B-GFP. Movie 1 shows a normal cell division in 10 min total time. Movie 2 and 3 show a division where the two daughter nuclei are connected by a thread of DNA. Movie 2 is taken in 86 frames collected at 16 sec time intervals (23 min total time). Movie 3 shows a cell division that lasts 20 minutes. The first 6 frames were collected at 30 sec intervals. The next 18 frames were collected at one minute intervals (total 20 min, 24 frames). These two daughter cells were observed for another 20 minutes at irregular time intervals, but no further significant movement was observed.
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Rens, W., Torosantucci, L., Degrassi, F. et al. Incomplete sister chromatid separation of long chromosome arms. Chromosoma 115, 481–490 (2006). https://doi.org/10.1007/s00412-006-0077-1
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DOI: https://doi.org/10.1007/s00412-006-0077-1