Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Mar 1;88(3):618–629. doi: 10.1083/jcb.88.3.618

A permeabilized cell model for studying cell division: a comparison of anaphase chromosome movement and cleavage furrow constriction in lysed PtK1 cells

PMCID: PMC2112770  PMID: 6111566

Abstract

After lysis in a Brij 58-polyethylene glycol medium, PtK1 cells are permeable to small molecules, such as erythrosin B, and to proteins, such as rhodamine-labeled FAB, myosin subfragment-1, and tubulin. Holes are present in the plasma membrane, and the mitochondria are swollen and distorted, but other membrane-bounded organelles of the lysed cell model are not noticeably altered. After lysis, the mitotic apparatus is functional; chromosomes move poleward and the spindle elongates. Cells lysed while in cytokinesis will continue to divide for several minutes. Addition of crude tubulin extracts, MAP-free tubulin, or taxol to the lysis medium retards anaphase chromosome movements but does not affect cleavage. On the other hand, N-ethylmaleimide-modified myosin subfragment-1, phalloidin, and cytochalasin B inhibit cleavage but have no effect on anaphase chromosome movements under identical lysis conditions. These results suggest that actomyosin plays no functional role in anaphase chromosome movement in mammalian tissue culture cells and that microtubule depolymerization is a rate-limiting step for chromosome-to-pole movements.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aubin J. E., Weber K., Osborn M. Analysis of actin and microfilament-associated proteins in the mitotic spindle and cleavage furrow of PtK2 cells by immunofluorescence microscopy. A critical note. Exp Cell Res. 1979 Nov;124(1):93–109. doi: 10.1016/0014-4827(79)90260-x. [DOI] [PubMed] [Google Scholar]
  2. Cande W. Z. A permeabilized cell model for studying cytokinesis using mammalian tissue culture cells. J Cell Biol. 1980 Nov;87(2 Pt 1):326–335. doi: 10.1083/jcb.87.2.326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cande W. Z., Lazarides E., McIntosh J. R. A comparison of the distribution of actin and tubulin in the mammalian mitotic spindle as seen by indirect immunofluorescence. J Cell Biol. 1977 Mar;72(3):552–567. doi: 10.1083/jcb.72.3.552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cande W. Z., Snyder J., Smith D., Summers K., McIntosh J. R. A functional mitotic spindle prepared from mammalian cells in culture. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1559–1563. doi: 10.1073/pnas.71.4.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cande W. Z., Wolniak S. M. Chromosome movement in lysed mitotic cells is inhibited by vanadate. J Cell Biol. 1978 Nov;79(2 Pt 1):573–580. doi: 10.1083/jcb.79.2.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dancker P., Löw I., Hasselbach W., Wieland T. Interaction of actin with phalloidin: polymerization and stabilization of F-actin. Biochim Biophys Acta. 1975 Aug 19;400(2):407–414. doi: 10.1016/0005-2795(75)90196-8. [DOI] [PubMed] [Google Scholar]
  7. Dietz R. Die Assembly-Hypothese der Chromosomenbewegung und die Veränderungen der Spindellänge während der Anaphase I in Spermatocyten von Pales ferruginea (Tipulidae, Diptera. Chromosoma. 1972;38(1):11–76. doi: 10.1007/BF00319955. [DOI] [PubMed] [Google Scholar]
  8. Fuge H. Ultrastructure of the mitotic spindle. Int Rev Cytol Suppl. 1977;(6):1–58. [PubMed] [Google Scholar]
  9. Fujiwara K., Pollard T. D. Fluorescent antibody localization of myosin in the cytoplasm, cleavage furrow, and mitotic spindle of human cells. J Cell Biol. 1976 Dec;71(3):848–875. doi: 10.1083/jcb.71.3.848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goodno C. C. Inhibition of myosin ATPase by vanadate ion. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2620–2624. doi: 10.1073/pnas.76.6.2620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Griffith L. M., Pollard T. D. Evidence for actin filament-microtubule interaction mediated by microtubule-associated proteins. J Cell Biol. 1978 Sep;78(3):958–965. doi: 10.1083/jcb.78.3.958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HOFFMANN-BERLING H. Die Bedeutung des Adenosintriphosphat für die Zell- und Kernteilungsbewegungen in der Anaphase. Biochim Biophys Acta. 1954 Oct;15(2):226–236. doi: 10.1016/0006-3002(54)90063-3. [DOI] [PubMed] [Google Scholar]
  13. HOFFMANN-BERLING H. Die glycerinwasserextrahierte Telophasezelle als Modell der Zytokinese. Biochim Biophys Acta. 1954 Nov;15(3):332–339. doi: 10.1016/0006-3002(54)90034-7. [DOI] [PubMed] [Google Scholar]
  14. Hartwig J. H., Stossel T. P. Cytochalasin B and the structure of actin gels. J Mol Biol. 1979 Nov 5;134(3):539–553. doi: 10.1016/0022-2836(79)90366-8. [DOI] [PubMed] [Google Scholar]
  15. Heppel L. A., Makan N. Methods for rapidly altering the permeability of mammalian cells. J Supramol Struct. 1977;6(3):399–409. doi: 10.1002/jss.400060313. [DOI] [PubMed] [Google Scholar]
  16. Mabuchi I., Okuno M. The effect of myosin antibody on the division of starfish blastomeres. J Cell Biol. 1977 Jul;74(1):251–263. doi: 10.1083/jcb.74.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Margolis R. L., Wilson L., Keifer B. I. Mitotic mechanism based on intrinsic microtubule behaviour. Nature. 1978 Mar 30;272(5652):450–452. doi: 10.1038/272450a0. [DOI] [PubMed] [Google Scholar]
  18. Meeusen R. L., Bennett J., Cande W. Z. Effect of microinjected N-ethylmaleimide-modified heavy meromyosin on cell division in amphibian eggs. J Cell Biol. 1980 Sep;86(3):858–865. doi: 10.1083/jcb.86.3.858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Meeusen R. L., Cande W. Z. N-ethylmaleimide-modified heavy meromyosin. A probe for actomyosin interactions. J Cell Biol. 1979 Jul;82(1):57–65. doi: 10.1083/jcb.82.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miller M. R., Castellot J. J., Jr, Pardee A. B. A general method for permeabilizing monolayer and suspension cultured animal cells. Exp Cell Res. 1979 May;120(2):421–425. doi: 10.1016/0014-4827(79)90404-x. [DOI] [PubMed] [Google Scholar]
  21. Nicklas R. B. Chromosome movement: current models and experiments on living cells. Soc Gen Physiol Ser. 1975;30:97–117. [PubMed] [Google Scholar]
  22. Nicklas R. B. Mitosis. Adv Cell Biol. 1971;2:225–297. doi: 10.1007/978-1-4615-9588-5_5. [DOI] [PubMed] [Google Scholar]
  23. Petzelt C. Biochemistry of the mitotic spindle. Int Rev Cytol. 1979;60:53–92. doi: 10.1016/s0074-7696(08)61259-0. [DOI] [PubMed] [Google Scholar]
  24. Sanger J. W. Mitosis in beating cardiac myoblasts treated with cytochalasin-B. J Exp Zool. 1977 Sep;201(3):463–469. doi: 10.1002/jez.1402010313. [DOI] [PubMed] [Google Scholar]
  25. Sanger J. W. Presence of actin during chromosomal movement. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2451–2455. doi: 10.1073/pnas.72.6.2451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schiff P. B., Horwitz S. B. Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1561–1565. doi: 10.1073/pnas.77.3.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schloss J. A., Milsted A., Goldman R. D. Myosin subfragment binding for the localization of actin-like microfilaments in cultured cells. A light and electron microscope study. J Cell Biol. 1977 Sep;74(3):794–815. doi: 10.1083/jcb.74.3.794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schroeder T. E. Dynamics of the contractile ring. Soc Gen Physiol Ser. 1975;30:305–334. [PubMed] [Google Scholar]
  29. Snyder J. A., McIntosh J. R. Initiation and growth of microtubules from mitotic centers in lysed mammalian cells. J Cell Biol. 1975 Dec;67(3):744–760. doi: 10.1083/jcb.67.3.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang Y. L., Taylor D. L. Distribution of fluorescently labeled actin in living sea urchin eggs during early development. J Cell Biol. 1979 Jun;81(3):672–679. doi: 10.1083/jcb.81.3.672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weeds A. G., Taylor R. S. Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin. Nature. 1975 Sep 4;257(5521):54–56. doi: 10.1038/257054a0. [DOI] [PubMed] [Google Scholar]
  32. Wehland J., Osborn M., Weber K. Phalloidin-induced actin polymerization in the cytoplasm of cultured cells interferes with cell locomotion and growth. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5613–5617. doi: 10.1073/pnas.74.12.5613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Weingarten M. D., Lockwood A. H., Hwo S. Y., Kirschner M. W. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. 1975 May;72(5):1858–1862. doi: 10.1073/pnas.72.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES