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Both actin and myosin inhibitors affect spindle architecture in PtK1 cells: does an actomyosin system contribute to mitotic spindle forces by regulating attachment and movements of chromosomes in mammalian cells?

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Abstract

Immunocytochemical techniques are used to analyze the effects of both an actin and myosin inhibitor on spindle architecture in PtK1 cells to understand why both these inhibitors slow or block chromosome motion and detach chromosomes. Cytochalasin J, an actin inhibitor and a myosin inhibitor, 2, 3 butanedione 2-monoxime, have similar effects on changes in spindle organization. Using primary antibodies and stains, changes are studied in microtubule (MT), actin, myosin, and chromatin localization. Treatment of mitotic cells with both inhibitors results in detachment or misalignment of chromosomes from the spindle and a prominent buckling of MTs within the spindle, particularly evident in kinetochore fibers. Evidence is presented to suggest that an actomyosin system may help to regulate the initial and continued attachment of chromosomes to the mammalian spindle and could also influence spindle checkpoint(s).

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Abbreviations

BDM:

2,3 butanedione 2-monoximine

CJ:

cytochalasin J

kMTs:

kinetochore microtubules

MTs:

microtubules

nkMTs:

nonkinetochore microtubules

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Acknowledgments

We would like to thank the Barton L. Weller Endowment to JAS for support for this research and Laura Daddow for reviewing this manuscript.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Denver, Denver, CO, 80208, USA

    Judith A. Snyder, Yen Ha, Claire Olsofka & Reema Wahdan

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  1. Judith A. Snyder
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  2. Yen Ha
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  3. Claire Olsofka
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  4. Reema Wahdan
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Correspondence to Judith A. Snyder.

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Snyder, J.A., Ha, Y., Olsofka, C. et al. Both actin and myosin inhibitors affect spindle architecture in PtK1 cells: does an actomyosin system contribute to mitotic spindle forces by regulating attachment and movements of chromosomes in mammalian cells?. Protoplasma 240, 57–68 (2010). https://doi.org/10.1007/s00709-009-0089-9

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  • DOI: https://doi.org/10.1007/s00709-009-0089-9

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