Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Mar 1;96(3):776–782. doi: 10.1083/jcb.96.3.776

Microtubule-organizing centers abnormal in number, structure, and nucleating activity in x-irradiated mammalian cells

PMCID: PMC2112387  PMID: 6833383

Abstract

Microtubule-organizing centers (MTOCs) in x-irradiated cells were visualized by immunofluorescence using antibody against tubulin. From two to ten reassembly sites of microtubules appeared after microtubule depolymerization at low temperature in an irradiated mitotic cell, in contrast to nonirradiated mitotic cells, which predominantly show 2 MTOCs. A time-course examination of MTOCs in synchronously cultured cells revealed that the multiple MTOCs appeared not immediately after irradiation but at the time of mitosis. Those multiple MTOCs formed at mitosis were inherited by the daughter cells in the next generation. The structure and capacity of the centrosomes to nucleate microtubules in vitro were then examined by electron microscopy of whole-mount preparations as well as by dark-field microscopy. About 70-80% of the centrosomes derived from nonirradiated cells were composed of a pair of centrioles and pericentriolar material, which initiated greater than 100 microtubules. The fraction of fully active complete centrosomes decreased with time of incubation after irradiation. These were replaced by disintegrated centrosomal components such as dissociated centrioles and pericentriolar cloud, a nucleating site with a single centriole, or only an amorphous structure of pericentriolar cloud. Assembly of less than 20 microtubules onto the amorphous cloud without centrioles was seen in 54% of the initiating sites in mitotic cells 2 d after irradiation. These results suggest that x-irradiation causes disintegration of centrosomes at mitosis when the structural and functional reorganization of centrosomes is believed to occur.

Full Text

The Full Text of this article is available as a PDF (2.0 MB).

Selected References

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

  1. Borisy G. G., Marcum J. M., Olmsted J. B., Murphy D. B., Johnson K. A. Purification of tubulin and associated high molecular weight proteins from porcine brain and characterization of microtubule assembly in vitro. Ann N Y Acad Sci. 1975 Jun 30;253:107–132. doi: 10.1111/j.1749-6632.1975.tb19196.x. [DOI] [PubMed] [Google Scholar]
  2. Brinkley B. R., Cox S. M., Pepper D. A., Wible L., Brenner S. L., Pardue R. L. Tubulin assembly sites and the organization of cytoplasmic microtubules in cultured mammalian cells. J Cell Biol. 1981 Sep;90(3):554–562. doi: 10.1083/jcb.90.3.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DIRKSEN E. R. The presence of centrioles in artificially activated sea urchin eggs. J Biophys Biochem Cytol. 1961 Oct;11:244–247. doi: 10.1083/jcb.11.1.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Brabander M., Geuens G., Nuydens R., Willebrords R., De Mey J. Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5608–5612. doi: 10.1073/pnas.78.9.5608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gould R. R., Borisy G. G. The pericentriolar material in Chinese hamster ovary cells nucleates microtubule formation. J Cell Biol. 1977 Jun;73(3):601–615. doi: 10.1083/jcb.73.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hurwitz C., Tolmach L. J. Time lapse cinemicrographic studies of x-irradiated HeLa S3 cells. I. Cell progression and cell disintegration. Biophys J. 1969 Apr;9(4):607–633. doi: 10.1016/S0006-3495(69)86407-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kuriyama R., Borisy G. G. Centriole cycle in Chinese hamster ovary cells as determined by whole-mount electron microscopy. J Cell Biol. 1981 Dec;91(3 Pt 1):814–821. doi: 10.1083/jcb.91.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kuriyama R., Borisy G. G. Microtubule-nucleating activity of centrosomes in Chinese hamster ovary cells is independent of the centriole cycle but coupled to the mitotic cycle. J Cell Biol. 1981 Dec;91(3 Pt 1):822–826. doi: 10.1083/jcb.91.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Marin G., Bender M. A. Radiation-induced mammalian cell death: lapse-time cinemicrographic observations. Exp Cell Res. 1966 Sep;43(2):413–423. doi: 10.1016/0014-4827(66)90068-1. [DOI] [PubMed] [Google Scholar]
  10. McGill M., Brinkley B. R. Human chromosomes and centrioles as nucleating sites for the in vitro assembly of microtubules from bovine brain tubulin. J Cell Biol. 1975 Oct;67(1):189–199. doi: 10.1083/jcb.67.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Osborn M., Weber K. Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane. Proc Natl Acad Sci U S A. 1976 Mar;73(3):867–871. doi: 10.1073/pnas.73.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sharp G. A., Osborn M., Weber K. Ultrastructure of multiple microtubule initiation sites in mouse neuroblastoma cells. J Cell Sci. 1981 Feb;47:1–24. doi: 10.1242/jcs.47.1.1. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Spiegelman B. M., Lopata M. A., Kirschner M. W. Aggregation of microtubule initiation sites preceding neurite outgrowth in mouse neuroblastoma cells. Cell. 1979 Feb;16(2):253–263. doi: 10.1016/0092-8674(79)90003-5. [DOI] [PubMed] [Google Scholar]
  15. Spiegelman B. M., Lopata M. A., Kirschner M. W. Multiple sites for the initiation of microtubule assembly in mammalian cells. Cell. 1979 Feb;16(2):239–252. doi: 10.1016/0092-8674(79)90002-3. [DOI] [PubMed] [Google Scholar]
  16. TERASIMA T., TOLMACH L. J. Growth and nucleic acid synthesis in synchronously dividing populations of HeLa cells. Exp Cell Res. 1963 Apr;30:344–362. doi: 10.1016/0014-4827(63)90306-9. [DOI] [PubMed] [Google Scholar]
  17. Telzer B. R., Rosenbaum J. L. Cell cycle-dependent, in vitro assembly of microtubules onto pericentriolar material of HeLa cells. J Cell Biol. 1979 Jun;81(3):484–497. doi: 10.1083/jcb.81.3.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Watt F. M., Harris H. Microtubule-organizing centres in mammalian cells in culture. J Cell Sci. 1980 Aug;44:103–121. doi: 10.1242/jcs.44.1.103. [DOI] [PubMed] [Google Scholar]
  19. Yakara I., Kakimoto-Sameshima F. Microtubule organization of lymphocytes and its modulation by patch and cap formation. Cell. 1978 Sep;15(1):251–259. doi: 10.1016/0092-8674(78)90100-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES