Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Dec 1;131(5):1125–1131. doi: 10.1083/jcb.131.5.1125

Chromosomes initiate spindle assembly upon experimental dissolution of the nuclear envelope in grasshopper spermatocytes

PMCID: PMC2120643  PMID: 8522577

Abstract

Chromosomes are known to enhance spindle microtubule assembly in grasshopper spermatocytes, which suggested to us that chromosomes might play an essential role in the initiation of spindle formation. Chromosomes might, for example, activate other spindle components such as centrosomes and tubulin subunits upon the breakdown of the nuclear envelope. We tested this possibility in living grasshopper spermatocytes. We ruptured the nuclear envelope during prophase, which prematurely exposed the centrosomes to chromosomes and nuclear sap. Spindle assembly was promptly initiated. In contrast, assembly of the spindle was completely inhibited if the nucleus was mechanically removed from a late prophase cell. Other experiments showed that the trigger for spindle assembly is associated with the chromosomes; other constituents of the nucleus cannot initiate spindle assembly in the absence of the chromosomes. The initiation of spindle assembly required centrosomes as well as chromosomes. Extracting centrosomes from late prophase cells completely inhibited spindle assembly after dissolution of the nuclear envelope. We conclude that the normal formation of a bipolar spindle in grasshopper spermatocytes is regulated by chromosomes. A possible explanation is an activator, perhaps a chromosomal protein (Yeo, J.-P., F. Alderuccio, and B.-H. Toh. 1994a. Nature (Lond.). 367: 288-291), that promotes and stabilizes the assembly of astral microtubules and thus promotes assembly of the spindle.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Aubin J. E., Osborn M., Weber K. Variations in the distribution and migration of centriole duplexes in mitotic PtK2 cells studied by immunofluorescence microscopy. J Cell Sci. 1980 Jun;43:177–194. doi: 10.1242/jcs.43.1.177. [DOI] [PubMed] [Google Scholar]
  2. Church K., Nicklas R. B., Lin H. P. Micromanipulated bivalents can trigger mini-spindle formation in Drosophila melanogaster spermatocyte cytoplasm. J Cell Biol. 1986 Dec;103(6 Pt 2):2765–2773. doi: 10.1083/jcb.103.6.2765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Inoué S., Sato H. Cell motility by labile association of molecules. The nature of mitotic spindle fibers and their role in chromosome movement. J Gen Physiol. 1967 Jul;50(6 Suppl):259–292. [PMC free article] [PubMed] [Google Scholar]
  4. Kallajoki M., Weber K., Osborn M. Ability to organize microtubules in taxol-treated mitotic PtK2 cells goes with the SPN antigen and not with the centrosome. J Cell Sci. 1992 May;102(Pt 1):91–102. doi: 10.1242/jcs.102.1.91. [DOI] [PubMed] [Google Scholar]
  5. Karsenti E., Newport J., Hubble R., Kirschner M. Interconversion of metaphase and interphase microtubule arrays, as studied by the injection of centrosomes and nuclei into Xenopus eggs. J Cell Biol. 1984 May;98(5):1730–1745. doi: 10.1083/jcb.98.5.1730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kiehart D. P. Microinjection of echinoderm eggs: apparatus and procedures. Methods Cell Biol. 1982;25(Pt B):13–31. doi: 10.1016/s0091-679x(08)61418-1. [DOI] [PubMed] [Google Scholar]
  7. Kirschner M., Mitchison T. Beyond self-assembly: from microtubules to morphogenesis. Cell. 1986 May 9;45(3):329–342. doi: 10.1016/0092-8674(86)90318-1. [DOI] [PubMed] [Google Scholar]
  8. Mazia D. The chromosome cycle and the centrosome cycle in the mitotic cycle. Int Rev Cytol. 1987;100:49–92. doi: 10.1016/s0074-7696(08)61698-8. [DOI] [PubMed] [Google Scholar]
  9. Mitchison T. J., Kirschner M. W. Properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation. J Cell Biol. 1985 Sep;101(3):766–777. doi: 10.1083/jcb.101.3.766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nicklas R. B., Kubai D. F., Hays T. S. Spindle microtubules and their mechanical associations after micromanipulation in anaphase. J Cell Biol. 1982 Oct;95(1):91–104. doi: 10.1083/jcb.95.1.91. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nicklas R. B., Kubai D. F. Microtubules, chromosome movement, and reorientation after chromosomes are detached from the spindle by micromanipulation. Chromosoma. 1985;92(4):313–324. doi: 10.1007/BF00329815. [DOI] [PubMed] [Google Scholar]
  12. Nicklas R. B., Ward S. C. Elements of error correction in mitosis: microtubule capture, release, and tension. J Cell Biol. 1994 Sep;126(5):1241–1253. doi: 10.1083/jcb.126.5.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Peter M., Nakagawa J., Dorée M., Labbé J. C., Nigg E. A. In vitro disassembly of the nuclear lamina and M phase-specific phosphorylation of lamins by cdc2 kinase. Cell. 1990 May 18;61(4):591–602. doi: 10.1016/0092-8674(90)90471-p. [DOI] [PubMed] [Google Scholar]
  14. Rattner J. B., Berns M. W. Distribution of microtubules during centriole separation in rat kangaroo (Potorous) cells. Cytobios. 1976;15(57):37–43. [PubMed] [Google Scholar]
  15. Rieder C. L., Alexander S. P. Kinetochores are transported poleward along a single astral microtubule during chromosome attachment to the spindle in newt lung cells. J Cell Biol. 1990 Jan;110(1):81–95. doi: 10.1083/jcb.110.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rieder C. L., Hard R. Newt lung epithelial cells: cultivation, use, and advantages for biomedical research. Int Rev Cytol. 1990;122:153–220. doi: 10.1016/s0074-7696(08)61208-5. [DOI] [PubMed] [Google Scholar]
  17. Roos U. P. Light and electron microscopy of rat kangaroo cells in mitosis. I. Formation and breakdown of the mitotic apparatus. Chromosoma. 1973;40(1):43–82. doi: 10.1007/BF00319836. [DOI] [PubMed] [Google Scholar]
  18. Salmon E. D. Spindle microtubules: thermodynamics of in vivo assembly and role in chromosome movement. Ann N Y Acad Sci. 1975 Jun 30;253:383–406. doi: 10.1111/j.1749-6632.1975.tb19216.x. [DOI] [PubMed] [Google Scholar]
  19. Sawin K. E., Mitchison T. J. Mitotic spindle assembly by two different pathways in vitro. J Cell Biol. 1991 Mar;112(5):925–940. doi: 10.1083/jcb.112.5.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sluder G., Miller F. J., Rieder C. L. The reproduction of centrosomes: nuclear versus cytoplasmic controls. J Cell Biol. 1986 Nov;103(5):1873–1881. doi: 10.1083/jcb.103.5.1873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sluder G., Rieder C. L. Experimental separation of pronuclei in fertilized sea urchin eggs: chromosomes do not organize a spindle in the absence of centrosomes. J Cell Biol. 1985 Mar;100(3):897–903. doi: 10.1083/jcb.100.3.897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Steffen W., Fuge H., Dietz R., Bastmeyer M., Müller G. Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation? J Cell Biol. 1986 May;102(5):1679–1687. doi: 10.1083/jcb.102.5.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Theurkauf W. E., Hawley R. S. Meiotic spindle assembly in Drosophila females: behavior of nonexchange chromosomes and the effects of mutations in the nod kinesin-like protein. J Cell Biol. 1992 Mar;116(5):1167–1180. doi: 10.1083/jcb.116.5.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vernos I., Raats J., Hirano T., Heasman J., Karsenti E., Wylie C. Xklp1, a chromosomal Xenopus kinesin-like protein essential for spindle organization and chromosome positioning. Cell. 1995 Apr 7;81(1):117–127. doi: 10.1016/0092-8674(95)90376-3. [DOI] [PubMed] [Google Scholar]
  25. Yeo J. P., Alderuccio F., Toh B. H. A new chromosomal protein essential for mitotic spindle assembly. Nature. 1994 Jan 20;367(6460):288–291. doi: 10.1038/367288a0. [DOI] [PubMed] [Google Scholar]
  26. Yeo J. P., Forer A., Toh B. H. A homologue of the human regulator of mitotic spindle assembly protein (RMSA-1) is present in crane fly and is associated with meiotic chromosomes. J Cell Sci. 1994 Jul;107(Pt 7):1845–1851. doi: 10.1242/jcs.107.7.1845. [DOI] [PubMed] [Google Scholar]
  27. Zhang D., Nicklas R. B. The impact of chromosomes and centrosomes on spindle assembly as observed in living cells. J Cell Biol. 1995 Jun;129(5):1287–1300. doi: 10.1083/jcb.129.5.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES