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Differences in centromere positioning of cycling and postmitotic human cell types

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Abstract

Centromere positioning in human cell nuclei was traced in non-cycling peripheral blood lymphocytes (G0) and in terminally differentiated monocytes, as well as in cycling phytohemagglutinin-stimulated lymphocytes, diploid lymphoblastoid cells, normal fibroblasts, and neuroblastoma SH-EP cells using immunostaining of kinetochores, confocal microscopy and three-dimensional image analysis. Cell cycle stages were identified for each individual cell by a combination of replication labeling with 5-bromo-2′-deoxyuridine and immunostaining of pKi67. We demonstrate that the behavior of centromeres is similar in all cell types studied: a large fraction of centromeres are in the nuclear interior during early G1; in late G1 and early S phase, centromeres shift to the nuclear periphery and fuse in clusters. Peripheral location and clustering of centromeres are most pronounced in non-cycling cells (G0) and terminally differentiated monocytes. In late S and G2, centromeres partially decluster and migrate towards the nuclear interior. In the rather flat nuclei of adherently growing fibroblasts and neuroblastoma cells, kinetochores showed asymmetrical distributions with preferential kinetochore location close either to the bottom side of the nucleus (adjacent to the growth surface) or to the nuclear upper side. This asymmetrical distribution of centromeres is considered to be a consequence of chromosome arrangement in anaphase rosettes.

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Acknowledgements

We are grateful to W.C. Earnshaw (University of Edinburgh) for the generous gift of anti-CENP-B and anti-CENP-C antibodies. B. Joffe (Technical University of Munich) has suggested and greatly helped with the analysis of the asymmetrical distribution of peripheral kinetochores in individual cells. This work was supported by grants from the Deutsche Forschungsgemeinschaft to T. Cremer (Cr 59/20, 1–3).

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Authors and Affiliations

  1. Department of Biology II, Humangenetik, Ludwig Maximillians University (LMU), Richard Wagner Str. 10, 80333, Munich, Germany

    Irina Solovei, Lothar Schermelleh, Klaus Düring, Andrea Engelhardt & Thomas Cremer

  2. Kirchhoff Institute of Physics, University of Heidelberg, 69120, Heidelberg, Germany

    Stefan Stein & Christoph Cremer

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  1. Irina Solovei
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  2. Lothar Schermelleh
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  3. Klaus Düring
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  4. Andrea Engelhardt
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  5. Stefan Stein
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  6. Christoph Cremer
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  7. Thomas Cremer
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Correspondence to Irina Solovei.

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Communicated by E.A. Nigg

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Solovei, I., Schermelleh, L., Düring, K. et al. Differences in centromere positioning of cycling and postmitotic human cell types. Chromosoma 112, 410–423 (2004). https://doi.org/10.1007/s00412-004-0287-3

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