Abstract
In interphase Chinese hamster ovary (CHO) cells, the centrosome is attached to the nucleus very firmly. This nuclear-centrosome complex is isolated as a coherent structure by lysis and extraction of cells with Triton X-100 in a low ionic strength medium. Under these conditions, the ultrastructure of the centrioles attached to the nucleus can be discerned by electron microscopy of whole-mount preparations. The structural changes of the centrioles as a function of the cell cycle were monitored by this technique. Specifically, centriolar profiles were placed into six categories according to their orientation and the length ratio of daughter and parent centrioles. The proportion of centrioles in each category was plotted as a frequency histogram. The morphological changes in the centriole cycle were characterized by three distinguishable events: nucleation, elongation, and disorientation. The progress of centrioles through these stages was determined in synchronous populations of cells starting from S or M phase, in cells inhibited in DNA synthesis by addition of thymidine, and in cytoplasts. The results provide a quantitative description of the events of the centriole cycle. They also show that, in complete cells, nucleation, elongation, and disorientation are not dependent upon DNA synthesis. However, in cytoplasts, although elongation and disorientation occur as in normal cells, nucleation is blocked. Procentriole formation appeared to be inhibited by the removal of the nucleus. We suggest that coordination of centriole replication and nuclear replication may depend upon a signal arising from the nucleus.
Full Text
The Full Text of this article is available as a PDF (900.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aaronson R. P., Blobel G. On the attachment of the nuclear pore complex. J Cell Biol. 1974 Sep;62(3):746–754. doi: 10.1083/jcb.62.3.746. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BOOTSMA D., BUDKE L., VOS O. STUDIES ON SYNCHRONOUS DIVISION OF TISSUE CULTURE CELLS INITIATED BY EXCESS THYMIDINE. Exp Cell Res. 1964 Jan;33:301–309. doi: 10.1016/s0014-4827(64)81035-1. [DOI] [PubMed] [Google Scholar]
- Bornens M. Is the centriole bound to the nuclear membrane? Nature. 1977 Nov 3;270(5632):80–82. doi: 10.1038/270080a0. [DOI] [PubMed] [Google Scholar]
- Connolly J. A., Kalnins V. I. Visualization of centrioles and basal bodies by fluorescent staining with nonimmune rabbit sera. J Cell Biol. 1978 Nov;79(2 Pt 1):526–532. doi: 10.1083/jcb.79.2.526. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- DeFoor P. H., Stubblefield E. Effects of actinomycin D, amethopterin, and 5-fluro-2'-deoxyuridine on procentriole formation in Chinese hamster fibroblasts in culture. Exp Cell Res. 1974 Mar 30;85(1):136–142. doi: 10.1016/0014-4827(74)90223-7. [DOI] [PubMed] [Google Scholar]
- Goldman R. D., Pollack R., Chang C. M., Bushnell A. Properties of enucleated cells. III. Changes in cytoplasmic architecture of enucleated BHK21 cells following trypsinization and replating. Exp Cell Res. 1975 Jun;93(1):175–183. doi: 10.1016/0014-4827(75)90437-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Kato K. H., Sugiyama M. On the de novo formation of the centriole in the activated sea urchin egg. Dev Growth Differ. 1971 Dec;13(4):359–366. doi: 10.1111/j.1440-169x.1971.00359.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Murray R. G., Murray A. S., Pizzo A. The fine structure of mitosis in rat thymic lymphocytes. J Cell Biol. 1965 Aug;26(2):601–619. doi: 10.1083/jcb.26.2.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nadezhdina E. S., Fais D., Chentsov Y. S. On the association of centrioles with the interphase nucleus. Eur J Cell Biol. 1979 Jun;19(2):109–115. [PubMed] [Google Scholar]
- Prescott D. M., Kirkpatrick J. B. Mass enucleation of cultured animal cells. Methods Cell Biol. 1973;7:189–202. doi: 10.1016/s0091-679x(08)61777-x. [DOI] [PubMed] [Google Scholar]
- Rattner J. B., Phillips S. G. Independence of centriole formation and DNA synthesis. J Cell Biol. 1973 May;57(2):359–372. doi: 10.1083/jcb.57.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robbins E., Jentzsch G., Micali A. The centriole cycle in synchronized HeLa cells. J Cell Biol. 1968 Feb;36(2):329–339. doi: 10.1083/jcb.36.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Starger J. M., Brown W. E., Goldman A. E., Goldman R. D. Biochemical and immunological analysis of rapidly purified 10-nm filaments from baby hamster kidney (BHK-21) cells. J Cell Biol. 1978 Jul;78(1):93–109. doi: 10.1083/jcb.78.1.93. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wheatley D. N. Pericentriolar virus-like particles in Chinese hamster ovary cells. J Gen Virol. 1974 Aug;24(2):395–399. doi: 10.1099/0022-1317-24-2-395. [DOI] [PubMed] [Google Scholar]
- Wise G. E., Prescott D. M. Ultrastructure of enucleated mammalian cells in culture. Exp Cell Res. 1973 Sep;81(1):63–72. doi: 10.1016/0014-4827(73)90111-0. [DOI] [PubMed] [Google Scholar]
- XEROS N. Deoxyriboside control and synchronization of mitosis. Nature. 1962 May 19;194:682–683. doi: 10.1038/194682a0. [DOI] [PubMed] [Google Scholar]