Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Dec 2;123(6):1475–1489. doi: 10.1083/jcb.123.6.1475

Interpolar spindle microtubules in PTK cells

PMCID: PMC2290872  PMID: 8253845

Abstract

Spindle microtubules (MTs) in PtK1 cells, fixed at stages from metaphase to telophase, have been reconstructed using serial sections, electron microscopy, and computer image processing. We have studied the class of MTs that form an interdigitating system connecting the two spindle poles (interpolar MTs or ipMTs) and their relationship to the spindle MTs that attach to kinetochores (kMTs). Viewed in cross section, the ipMTs cluster with antiparallel near neighbors throughout mitosis; this bundling becomes much more pronounced as anaphase proceeds. While the minus ends of most kMTs are near the poles, those of the ipMTs are spread over half of the spindle length, with at least 50% lying > 1.5 microns from the poles. Longitudinal views of the ipMT bundles demonstrate a major rearrangement of their plus ends between mid- and late anaphase B. However, the minus ends of these MTs do not move appreciably farther from the spindle midplane, suggesting that sliding of these MTs contributes little to anaphase B. The minus ends of ipMTs are markedly clustered in the bundles of kMTs throughout anaphase A. These ends lie close to kMTs much more frequently than would be expected by chance, suggesting a specific interaction. As sister kinetochores separate and kMTs shorten, the minus ends of the kMTs remain associated with the spindle poles, but the minus ends of many ipMTs are released from the kMT bundles, allowing the spindle pole and the kMTs to move away from the ipMTs as the spindle elongates.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

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

  1. Aist J. R., Bayles C. J., Tao W., Berns M. W. Direct experimental evidence for the existence, structural basis and function of astral forces during anaphase B in vivo. J Cell Sci. 1991 Oct;100(Pt 2):279–288. doi: 10.1242/jcs.100.2.279. [DOI] [PubMed] [Google Scholar]
  2. Aist J. R., Berns M. W. Mechanics of chromosome separation during mitosis in Fusarium (Fungi imperfecti): new evidence from ultrastructural and laser microbeam experiments. J Cell Biol. 1981 Nov;91(2 Pt 1):446–458. doi: 10.1083/jcb.91.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aist J. R., Liang H., Berns M. W. Astral and spindle forces in PtK2 cells during anaphase B: a laser microbeam study. J Cell Sci. 1993 Apr;104(Pt 4):1207–1216. doi: 10.1242/jcs.104.4.1207. [DOI] [PubMed] [Google Scholar]
  4. Bajer A. S. Functional autonomy of monopolar spindle and evidence for oscillatory movement in mitosis. J Cell Biol. 1982 Apr;93(1):33–48. doi: 10.1083/jcb.93.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bajer A. S., Molè-Bajer J. Reorganization of microtubules in endosperm cells and cell fragments of the higher plant Haemanthus in vivo. J Cell Biol. 1986 Jan;102(1):263–281. doi: 10.1083/jcb.102.1.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bastmeyer M., Fuge H. The distribution of intermicrotubular bridges in meiotic spindles of the crane fly. Chromosoma. 1986;94(5):419–424. doi: 10.1007/BF00328643. [DOI] [PubMed] [Google Scholar]
  7. Belmont L. D., Hyman A. A., Sawin K. E., Mitchison T. J. Real-time visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts. Cell. 1990 Aug 10;62(3):579–589. doi: 10.1016/0092-8674(90)90022-7. [DOI] [PubMed] [Google Scholar]
  8. Brinkley B. R., Cartwright J., Jr Ultrastructural analysis of mitotic spindle elongation in mammalian cells in vitro. Direct microtubule counts. J Cell Biol. 1971 Aug;50(2):416–431. doi: 10.1083/jcb.50.2.416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ding R., McDonald K. L., McIntosh J. R. Three-dimensional reconstruction and analysis of mitotic spindles from the yeast, Schizosaccharomyces pombe. J Cell Biol. 1993 Jan;120(1):141–151. doi: 10.1083/jcb.120.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fuge H. The arrangement of microtubules and the attachment of chromosomes to the spindle during anaphase in tipulid spermatocytes. Chromosoma. 1974 Apr 9;45(3):245–260. doi: 10.1007/BF00283409. [DOI] [PubMed] [Google Scholar]
  11. Fuge H. The three-dimensional architecture of chromosome fibres in the crane fly. II. Amphitelic sex univalents in meiotic anaphase I. Chromosoma. 1985;91(3-4):322–328. doi: 10.1007/BF00328228. [DOI] [PubMed] [Google Scholar]
  12. Heath I. B. Mitosis in the fungus Thraustotheca clavata. J Cell Biol. 1974 Jan;60(1):204–220. doi: 10.1083/jcb.60.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hepler P. K., Jackson W. T. Microtubules and early stages of cell-plate formation in the endosperm of Haemanthus katherinae Baker. J Cell Biol. 1968 Aug;38(2):437–446. doi: 10.1083/jcb.38.2.437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hiramoto Y., Nakano Y. Micromanipulation studies of the mitotic apparatus in sand dollar eggs. Cell Motil Cytoskeleton. 1988;10(1-2):172–184. doi: 10.1002/cm.970100122. [DOI] [PubMed] [Google Scholar]
  15. Hogan C. J., Cande W. Z. Antiparallel microtubule interactions: spindle formation and anaphase B. Cell Motil Cytoskeleton. 1990;16(2):99–103. doi: 10.1002/cm.970160203. [DOI] [PubMed] [Google Scholar]
  16. Horio T., Hotani H. Visualization of the dynamic instability of individual microtubules by dark-field microscopy. Nature. 1986 Jun 5;321(6070):605–607. doi: 10.1038/321605a0. [DOI] [PubMed] [Google Scholar]
  17. Julian M., Tollon Y., Lajoie-Mazenc I., Moisand A., Mazarguil H., Puget A., Wright M. gamma-Tubulin participates in the formation of the midbody during cytokinesis in mammalian cells. J Cell Sci. 1993 May;105(Pt 1):145–156. doi: 10.1242/jcs.105.1.145. [DOI] [PubMed] [Google Scholar]
  18. Masuda H., Hirano T., Yanagida M., Cande W. Z. In vitro reactivation of spindle elongation in fission yeast nuc2 mutant cells. J Cell Biol. 1990 Feb;110(2):417–425. doi: 10.1083/jcb.110.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Masuda H., McDonald K. L., Cande W. Z. The mechanism of anaphase spindle elongation: uncoupling of tubulin incorporation and microtubule sliding during in vitro spindle reactivation. J Cell Biol. 1988 Aug;107(2):623–633. doi: 10.1083/jcb.107.2.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McDonald K. L., Edwards M. K., McIntosh J. R. Cross-sectional structure of the central mitotic spindle of Diatoma vulgare. Evidence for specific interactions between antiparallel microtubules. J Cell Biol. 1979 Nov;83(2 Pt 1):443–461. doi: 10.1083/jcb.83.2.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McDonald K. L., O'Toole E. T., Mastronarde D. N., McIntosh J. R. Kinetochore microtubules in PTK cells. J Cell Biol. 1992 Jul;118(2):369–383. doi: 10.1083/jcb.118.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McIntosh J. R., Cande W. Z., Snyder J. A. Structure and physiology of the mammalian mitotic spindle. Soc Gen Physiol Ser. 1975;30:31–76. [PubMed] [Google Scholar]
  23. McIntosh J. R., McDonald K. L., Edwards M. K., Ross B. M. Three-dimensional structure of the central mitotic spindle of Diatoma vulgare. J Cell Biol. 1979 Nov;83(2 Pt 1):428–442. doi: 10.1083/jcb.83.2.428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McIntosh J. R., Roos U. P., Neighbors B., McDonald K. L. Architecture of the microtubule component of mitotic spindles from Dictyostelium discoideum. J Cell Sci. 1985 Apr;75:93–129. doi: 10.1242/jcs.75.1.93. [DOI] [PubMed] [Google Scholar]
  25. Mitchison T. J. Polewards microtubule flux in the mitotic spindle: evidence from photoactivation of fluorescence. J Cell Biol. 1989 Aug;109(2):637–652. doi: 10.1083/jcb.109.2.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mitchison T. J., Salmon E. D. Poleward kinetochore fiber movement occurs during both metaphase and anaphase-A in newt lung cell mitosis. J Cell Biol. 1992 Nov;119(3):569–582. doi: 10.1083/jcb.119.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nislow C., Lombillo V. A., Kuriyama R., McIntosh J. R. A plus-end-directed motor enzyme that moves antiparallel microtubules in vitro localizes to the interzone of mitotic spindles. Nature. 1992 Oct 8;359(6395):543–547. doi: 10.1038/359543a0. [DOI] [PubMed] [Google Scholar]
  28. Oakley B. R., Oakley C. E., Yoon Y., Jung M. K. Gamma-tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans. Cell. 1990 Jun 29;61(7):1289–1301. doi: 10.1016/0092-8674(90)90693-9. [DOI] [PubMed] [Google Scholar]
  29. Paweletz N. Zur Funktion des "Flemming-Körpers" bei der Teilung tierischer Zellen. Naturwissenschaften. 1967 Oct;54(20):533–535. doi: 10.1007/BF00627210. [DOI] [PubMed] [Google Scholar]
  30. Pfarr C. M., Coue M., Grissom P. M., Hays T. S., Porter M. E., McIntosh J. R. Cytoplasmic dynein is localized to kinetochores during mitosis. Nature. 1990 May 17;345(6272):263–265. doi: 10.1038/345263a0. [DOI] [PubMed] [Google Scholar]
  31. Sawin K. E., LeGuellec K., Philippe M., Mitchison T. J. Mitotic spindle organization by a plus-end-directed microtubule motor. Nature. 1992 Oct 8;359(6395):540–543. doi: 10.1038/359540a0. [DOI] [PubMed] [Google Scholar]
  32. Saxton W. M., McIntosh J. R. Interzone microtubule behavior in late anaphase and telophase spindles. J Cell Biol. 1987 Aug;105(2):875–886. doi: 10.1083/jcb.105.2.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schroeder T. E. The contractile ring. I. Fine structure of dividing mammalian (HeLa) cells and the effects of cytochalasin B. Z Zellforsch Mikrosk Anat. 1970;109(4):431–449. [PubMed] [Google Scholar]
  34. Shelden E., Wadsworth P. Interzonal microtubules are dynamic during spindle elongation. J Cell Sci. 1990 Oct;97(Pt 2):273–281. doi: 10.1242/jcs.97.2.273. [DOI] [PubMed] [Google Scholar]
  35. Smirnova E. A., Bajer A. S. Spindle poles in higher plant mitosis. Cell Motil Cytoskeleton. 1992;23(1):1–7. doi: 10.1002/cm.970230102. [DOI] [PubMed] [Google Scholar]
  36. Steffen W. Relationship between the arrangement of microtubules and chromosome behaviour of syntelic autosomal univalents during prometaphase in crane fly spermatocytes. Chromosoma. 1986;94(5):412–418. doi: 10.1007/BF00328642. [DOI] [PubMed] [Google Scholar]
  37. Steuer E. R., Wordeman L., Schroer T. A., Sheetz M. P. Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature. 1990 May 17;345(6272):266–268. doi: 10.1038/345266a0. [DOI] [PubMed] [Google Scholar]
  38. Tippit D. H., Fields C. T., O'Donnell K. L., Pickett-Heaps J. D., McLaughlin D. J. The organization of microtubules during anaphase and telophase spindle elongation in the rust fungus Puccinia. Eur J Cell Biol. 1984 May;34(1):34–44. [PubMed] [Google Scholar]
  39. Tippit D. H., Pillus L., Pickett-Heaps J. D. Near-neighbor analysis of spindle microtubules in the alga Ochromonas. Eur J Cell Biol. 1983 Mar;30(1):9–17. [PubMed] [Google Scholar]
  40. Tippit D. H., Pillus L., Pickett-Heaps J. Organization of spindle microtubules in Ochromonas danica. J Cell Biol. 1980 Dec;87(3 Pt 1):531–545. doi: 10.1083/jcb.87.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tippit D. H., Schulz D., Pickett-Heaps J. D. Analysis of the distribution of spindle microtubules in the diatom Fragilaria. J Cell Biol. 1978 Dec;79(3):737–763. doi: 10.1083/jcb.79.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Waters J. C., Cole R. W., Rieder C. L. The force-producing mechanism for centrosome separation during spindle formation in vertebrates is intrinsic to each aster. J Cell Biol. 1993 Jul;122(2):361–372. doi: 10.1083/jcb.122.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES