Abstract
We have microinjected aphidicolin, a specific inhibitor of DNA polymerase alpha, into syncytial Drosophila embryos. This treatment inhibits DNA synthesis and, as a consequence, nuclear replication. We demonstrate that under these conditions several cycles of both centrosome replication and cortical budding continue, although the cycles have a longer periodicity than is normally found. As in uninjected embryos, when the cortical buds are present, the embryos have nuclei containing decondensed chromatin surrounded by nuclear membranes as judged by bright annular staining with an anti-lamin antibody. As the buds recede, the unreplicated chromatin condenses and lamin staining becomes weak and diffuse. Thus, both cytoplasmic and nuclear aspects of the mitotic cycle continue following the inhibition of DNA replication in the Drosophila embryo.
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- Brachet J., de Petrocellis B. The effects of aphidicolin, an inhibitor of DNA replication, on sea urchin development. Exp Cell Res. 1981 Sep;135(1):179–189. doi: 10.1016/0014-4827(81)90310-4. [DOI] [PubMed] [Google Scholar]
- Dabauvalle M. C., Doree M., Bravo R., Karsenti E. Role of nuclear material in the early cell cycle of Xenopus embryos. Cell. 1988 Feb 26;52(4):525–533. doi: 10.1016/0092-8674(88)90465-5. [DOI] [PubMed] [Google Scholar]
- Evans T., Rosenthal E. T., Youngblom J., Distel D., Hunt T. Cyclin: a protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division. Cell. 1983 Jun;33(2):389–396. doi: 10.1016/0092-8674(83)90420-8. [DOI] [PubMed] [Google Scholar]
- Foe V. E., Alberts B. M. Studies of nuclear and cytoplasmic behaviour during the five mitotic cycles that precede gastrulation in Drosophila embryogenesis. J Cell Sci. 1983 May;61:31–70. doi: 10.1242/jcs.61.1.31. [DOI] [PubMed] [Google Scholar]
- Freeman M., Glover D. M. The gnu mutation of Drosophila causes inappropriate DNA synthesis in unfertilized and fertilized eggs. Genes Dev. 1987 Nov;1(9):924–930. doi: 10.1101/gad.1.9.924. [DOI] [PubMed] [Google Scholar]
- Freeman M., Nüsslein-Volhard C., Glover D. M. The dissociation of nuclear and centrosomal division in gnu, a mutation causing giant nuclei in Drosophila. Cell. 1986 Aug 1;46(3):457–468. doi: 10.1016/0092-8674(86)90666-5. [DOI] [PubMed] [Google Scholar]
- Fuchs J. P., Giloh H., Kuo C. H., Saumweber H., Sedat J. Nuclear structure: determination of the fate of the nuclear envelope in Drosophila during mitosis using monoclonal antibodies. J Cell Sci. 1983 Nov;64:331–349. doi: 10.1242/jcs.64.1.331. [DOI] [PubMed] [Google Scholar]
- Gerhart J., Wu M., Kirschner M. Cell cycle dynamics of an M-phase-specific cytoplasmic factor in Xenopus laevis oocytes and eggs. J Cell Biol. 1984 Apr;98(4):1247–1255. doi: 10.1083/jcb.98.4.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hara K., Tydeman P., Kirschner M. A cytoplasmic clock with the same period as the division cycle in Xenopus eggs. Proc Natl Acad Sci U S A. 1980 Jan;77(1):462–466. doi: 10.1073/pnas.77.1.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ikegami S., Amemiya S., Oguro M., Nagano H., Mano Y. Inhibition by aphidicolin of cell cycle progression and DNA replication in sea urchin embryos. J Cell Physiol. 1979 Sep;100(3):439–444. doi: 10.1002/jcp.1041000307. [DOI] [PubMed] [Google Scholar]
- Ikegami S., Taguchi T., Ohashi M., Oguro M., Nagano H., Mano Y. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha. Nature. 1978 Oct 5;275(5679):458–460. doi: 10.1038/275458a0. [DOI] [PubMed] [Google Scholar]
- Kilmartin J. V., Wright B., Milstein C. Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol. 1982 Jun;93(3):576–582. doi: 10.1083/jcb.93.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kimelman D., Kirschner M., Scherson T. The events of the midblastula transition in Xenopus are regulated by changes in the cell cycle. Cell. 1987 Feb 13;48(3):399–407. doi: 10.1016/0092-8674(87)90191-7. [DOI] [PubMed] [Google Scholar]
- Nagano H., Hirai S., Okano K., Ikegami S. Achromosomal cleavage of fertilized starfish eggs in the presence of aphidicolin. Dev Biol. 1981 Jul 30;85(2):409–415. doi: 10.1016/0012-1606(81)90272-4. [DOI] [PubMed] [Google Scholar]
- Nishioka D., Balczon R., Schatten G. Relationships between DNA synthesis and mitotic events in fertilized sea urchin eggs: aphidicolin inhibits DNA synthesis, nuclear breakdown and proliferation of microtubule organizing centers, but not cycles of microtubule assembly. Cell Biol Int Rep. 1984 Apr;8(4):337–346. doi: 10.1016/0309-1651(84)90161-9. [DOI] [PubMed] [Google Scholar]
- Rosenthal E. T., Hunt T., Ruderman J. V. Selective translation of mRNA controls the pattern of protein synthesis during early development of the surf clam, Spisula solidissima. Cell. 1980 Jun;20(2):487–494. doi: 10.1016/0092-8674(80)90635-2. [DOI] [PubMed] [Google Scholar]
- Sluder G., Begg D. A. Control mechanisms of the cell cycle: role of the spatial arrangement of spindle components in the timing of mitotic events. J Cell Biol. 1983 Sep;97(3):877–886. doi: 10.1083/jcb.97.3.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sluder G., Lewis K. Relationship between nuclear DNA synthesis and centrosome reproduction in sea urchin eggs. J Exp Zool. 1987 Oct;244(1):89–100. doi: 10.1002/jez.1402440111. [DOI] [PubMed] [Google Scholar]
- 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]
- Sluder G. Role of spindle microtubules in the control of cell cycle timing. J Cell Biol. 1979 Mar;80(3):674–691. doi: 10.1083/jcb.80.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Standart N., Minshull J., Pines J., Hunt T. Cyclin synthesis, modification and destruction during meiotic maturation of the starfish oocyte. Dev Biol. 1987 Nov;124(1):248–258. doi: 10.1016/0012-1606(87)90476-3. [DOI] [PubMed] [Google Scholar]
- Warn R. M., Flegg L., Warn A. An investigation of microtubule organization and functions in living Drosophila embryos by injection of a fluorescently labeled antibody against tyrosinated alpha-tubulin. J Cell Biol. 1987 Oct;105(4):1721–1730. doi: 10.1083/jcb.105.4.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warn R. M., Magrath R., Webb S. Distribution of F-actin during cleavage of the Drosophila syncytial blastoderm. J Cell Biol. 1984 Jan;98(1):156–162. doi: 10.1083/jcb.98.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whitfield W. G., Millar S. E., Saumweber H., Frasch M., Glover D. M. Cloning of a gene encoding an antigen associated with the centrosome in Drosophila. J Cell Sci. 1988 Apr;89(Pt 4):467–480. doi: 10.1242/jcs.89.4.467. [DOI] [PubMed] [Google Scholar]