, Volume 92, Issue 4, pp 313–324 | Cite as

Microtubules, chromosome movement, and reorientation after chromosomes are detached from the spindle by micromanipulation

  • R. Bruce Nicklas
  • Donna F. Kubai


The relationship between chromosome movement and mirotubules was explored by combining micromanipulation of living grasshopper spermatocytes with electron microscopy. We detached chromosomes from the spindle and placed them far out in the cytoplasm. Soon, the chromosomes began to move back toward the spindle and the cells were fixed at a chosen moment. The microtubules seen in three-dimensional reconstructions were correlated with the chromosome movement just prior to fixation. Before movement began, detached chromosomes had no kinetochore microtubules or a single one at most. Renewed movement was always accompanied by the reappearance of kinetochore microtubules; a single kinetochore microtubule appeared to suffice. Chromosome movements and kinetochore microtubule arrangements were unusual after reattachment, but their relationship was not: poleward forces, parallel to the kinetochore microtubule axis (as in normal anaphase), would explain the movement, however odd. The initial arrangement of kinetochore microtubules would have led to aberrant chromosome distribution if it persisted, but instead, reorientation to the appropriate arrangement always followed. Observations on living cells permitted us to place in sequence the kinetochore microtubule arrangements seen in fixed cells, revealing the microtubule transformations during reorientation. From the sequence of events we conclude that chromosome movement can cause reorientation to begin and that in the changes which follow, an unstable attachment of kinetochore microtubules to the spindle plays a major role.


Electron Microscopy Living Cell Developmental Biology Chromosome Distribution Fixed Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Begg DA (1975) Chromosome movement following detachment from the meiotic spindle by micromanipulation. J Cell Biol 67:25aGoogle Scholar
  2. Begg DA, Ellis GW (1979a) Micromanipulation studies of chromosome movement. I. Chromosome-spindle attachment and the mechanical properties of chromosomal spindle fibers. J Cell Biol 82:528–541Google Scholar
  3. Begg DA, Ellis GW (1979b) Micromanipulation studies of chromosome movement. II. Birefringent chromosomal fibers and the mechanical attachment of chromosomes to the spindle. J Cell Biol 82:542–554Google Scholar
  4. Church K, Lin H-P (1982) Meiosis in Drosophila melanogaster. II. The prometaphase-I kinetochore microtubule bundle and kinetochore orientation in males. J Cell Biol 93:365–373Google Scholar
  5. Dietz R (1956) Die Spermatocytenteilungen der Tipuliden. II. Graphische Analyse der Chromosomenbewegung während der Prometaphase I im Leben. Chromosoma 8:183–211Google Scholar
  6. Dietz R (1958) Multiple Geschlechtschromosomen bei den cypriden Ostracoden, ihre Evolution und ihre Teilungsverhalten. Chromosoma 9:359–440Google Scholar
  7. Margolis RL, Wilson L (1981) Microtubule treadmills — Possible molecular machinery. Nature 293:705–711Google Scholar
  8. McIntosh JR, Euteneuer U (1984) Tubulin hooks as probes for microtubule polarity: An analysis of the method and an evaluation of data on microtubule polarity in the mitotic spindle. J Cell Biol 98:525–533Google Scholar
  9. Mitchison T, Kirschner M (1984) Microtubule dynamics and cellular morphogenesis. In: Borisy GG, Cleveland DW, Murphy DB (eds) Molecular biology of the cytoskeleton. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 27–44Google Scholar
  10. Mitchison T, Kirschner M (1985) The properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation. J Cell Biol (in press)Google Scholar
  11. Moens PB, Moens T (1981) Computer measurements and graphics of three-dimensional ultrastructure. J Ultrastruct Res 75:131–141Google Scholar
  12. Nicklas RB (1967) Chromosome micromanipulation. II. Induced reorientation and the experimental control of segregation in meiosis. Chromosoma 21:17–50Google Scholar
  13. Nicklas RB (1971) Mitosis. In: Prescott DM, Goldstein L, McConkey E (eds) Advances in cell biology, vol 2. Appleton-Century-Crofts New York, pp 225–297Google Scholar
  14. Nicklas RB (1977) Chromosome distribution: Experiments on cell hybrids and in vitro. Phil Trans R Soc Lond B227:267–276Google Scholar
  15. Nicklas RB (1983) Measurements of the force produced by the mitotic spindle in anaphase. J Cell Biol 97:542–548Google Scholar
  16. Nicklas RB, Gordon GW (1985) The total length of spindle microtubules depends on the number of chromosomes present. J Cell Biol 100:1–7Google Scholar
  17. Nicklas RB, Staehly CA (1967) Chromosome micromanipulation. I. The mechanics of chromosome attachment to the spindle. Chromosoma 21:1–16Google Scholar
  18. Nicklas RB, Brinkley BR, Pepper DA, Kubai DF, Rickards GK (1979) Electron microscopy of spermatocytes previously studied in life: Methods and some observations on micromanipulated chromosomes. J Cell Sci 35:87–104Google Scholar
  19. Nicklas RB, Kubai DF, Hays TS (1982) Spindle microtubules and their mechanical associations after micromanipulation in anaphase. J Cell Biol 95:91–104Google Scholar
  20. Östergren G (1951) The mechanism of co-orientation in bivalents and multivalents. Hereditas 37:85–156Google Scholar
  21. Rieder CL (1982) The formation, structure, and composition of the mammalian kinetochore and kinetochore fiber. Int Rev Cytol 79:1–58Google Scholar
  22. Salmon ED, Leslie RJ, Saxton WM, Karow ML, McIntosh JR (1984) Spindle microtubule dynamics in sea urchin embryos: Analysis using a fluorescein-labeled tubulin and measurements of fluorescence redistribution after laser photobleaching. J Cell Biol 99:2165–2174Google Scholar
  23. Solari AJ, Counce SJ (1977) Synaptonemal complex karyotyping in Melanoplus differentialis. J Cell Sci 26:229–250Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • R. Bruce Nicklas
    • 1
  • Donna F. Kubai
    • 1
  1. 1.Department of ZoologyDuke UniversityDurhamUSA

Personalised recommendations