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Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts.
Open accessAbstract
Incubation of demembranated sperm chromatin in cytoplasmic extracts of unfertilized Xenopus laevis eggs resulted in nuclear envelope assembly, chromosome decondensation, and sperm pronuclear formation. In contrast, egg extracts made with EGTA-containing buffers induced the sperm chromatin to form chromosomes or irregularly shaped clumps of chromatin that were incorporated into bipolar or multipolar spindles. The 150,000 g supernatants of the EGTA extracts could not alone support these changes in incubated nuclei. However, these supernatants induced not only chromosome condensation and spindle formation, but also nuclear envelope breakdown when added to sperm pronuclei or isolated Xenopus liver or brain nuclei that were incubated in extracts made without EGTA. Similar changes were induced by partially purified preparations of maturation-promoting factor. The addition of calcium chloride to extracts containing condensed chromosomes and spindles caused dissolution of the spindles, decondensation of the chromosomes, and re-formation of interphase nuclei. These results indicate that nuclear envelope breakdown, chromosome condensation, and spindle assembly, as well as the regulation of these processes by Ca2+-sensitive cytoplasmic components, can be studied in vitro using extracts of amphibian eggs.
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Cited by
Chapter 30 Cell Cycle Extracts
Murray, A. W. (1991) · Methods in Cell Biology · doi:10.1016/s0091-679x(08)60298-8 · Cited by 953
Blangy, A., Lane, H. A., d'Hérin, P., Harper, M., Kress, M., & Niggt, E. A. (1995) · Cell · doi:10.1016/0092-8674(95)90142-6 · Cited by 952
Gautier, J., Norbury, C., Lohka, M., Nurse, P., & Maller, J. (1988) · Cell · doi:10.1016/0092-8674(88)90206-1 · Cited by 871
Self-citationThe Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis
Dunphy, W. G., Brizuela, L., Beach, D., & Newport, J. (1988) · Cell · doi:10.1016/0092-8674(88)90205-x · Cited by 816
Association of Spindle Assembly Checkpoint Component XMAD2 with Unattached Kinetochores
Chen, R.-H., Waters, J. C., Salmon, E. D., & Murray, A. W. (1996) · Science · doi:10.1126/science.274.5285.242 · Cited by 783
Metadata
- Authors
- Lohka, M. J., & Maller, J. L.
- Venue
- The Journal of cell biology
- Publisher
- Rockefeller University Press
- Published
- 1985-08-01
- Volume
- 101
- Issue
- 2
- Pages
- 518-523
- ISSN
- 0021-9525 (ISSN-L)
- 1540-8140
- OpenAlex
- W2013801092
- Cited by
- 404
- References
- 0
- FWCI
- 8.34
- Type
- journal article
- Language
- en
- Open access
- green
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- Lohka, M. J., & Maller, J. L. (1985). Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts. The Journal of cell biology. https://doi.org/10.1083/jcb.101.2.518
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