Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Apr 1;98(4):1247–1255. doi: 10.1083/jcb.98.4.1247

Cell cycle dynamics of an M-phase-specific cytoplasmic factor in Xenopus laevis oocytes and eggs

PMCID: PMC2113233  PMID: 6425302

Abstract

We have examined the regulation of maturation-promoting factor (MPF) activity in the mitotic and meiotic cell cycles of Xenopus laevis eggs and oocytes. To this end, we developed a method for the small scale extraction of eggs and oocytes and measured MPF activity in extracts by a dilution end point assay. We find that in oocytes, MPF activity appears before germinal vesicle breakdown and then disappears rapidly at the end of the first meiotic cycle. In the second meiotic cycle, MPF reappears before second metaphase, when maturation arrests. Thus, MPF cycling coincides with the abbreviated cycles of meiosis. When oocytes are induced to mature by low levels of injected MPF, cycloheximide does not prevent the appearance of MPF at high levels in the first cycle. This amplification indicates that an MPF precursor is present in the oocyte and activated by posttranslational means, triggered by the low level of injected MPF. Furthermore, MPF disappears approximately on time in such oocytes, indicating that the agent for MPF inactivation is also activated by posttranslational means. However, in the absence of protein synthesis, MPF never reappears in the second meiotic cycle. Upon fertilization or artificial activation of normal eggs, MPF disappears from the cytoplasm within 8 min. For a period thereafter, the inactivating agent remains able to destroy large amounts of MPF injected into the egg. It loses activity just as endogenous MPF appears at prophase of the first mitotic cycle. The repeated reciprocal cycling of MPF and the inactivating agent during cleavage stages is unaffected by colchicine and nocodazole and therefore does not require the effective completion of spindle formation, mitosis, or cytokinesis. However, MPF appearance is blocked by cycloheximide applied before mitosis; and MPF disappearance is blocked by cytostatic factor. In all these respects, MPF and the inactivating agent seem to be tightly linked to, and perhaps participate in, the cell cycle oscillator previously described for cleaving eggs of Xenopus laevis (Hara, K., P. Tydeman, and M. Kirschner, 1980, Proc. Natl. Acad. Sci. USA, 77:462- 466).

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Brachet J., Hanocq F., Van Gansen P. A cytochemical and ultrastructural analysis of in vitro maturation in amphibian oocytes. Dev Biol. 1970 Feb;21(1):157–195. doi: 10.1016/0012-1606(70)90067-9. [DOI] [PubMed] [Google Scholar]
  2. Dorée M. Protein synthesis is not involved in initiation or amplification of the maturation-promoting factor (MPF) in starfish oocytes. Exp Cell Res. 1982 May;139(1):127–133. doi: 10.1016/0014-4827(82)90326-3. [DOI] [PubMed] [Google Scholar]
  3. Drury K. C., Schorderet-Slatkine S. Effects of cycloheximide on the "autocatalytic" nature of the maturation promoting factor (MPF) in oocytes of Xenopus laevis. Cell. 1975 Mar;4(3):269–274. doi: 10.1016/0092-8674(75)90175-0. [DOI] [PubMed] [Google Scholar]
  4. Drury K. Method for the preparation of active maturation promoting factor (MPF) from in vitro matured oocytes of Xenopus laevis. Differentiation. 1978 May 26;10(3):181–186. doi: 10.1111/j.1432-0436.1978.tb00962.x. [DOI] [PubMed] [Google Scholar]
  5. Gerhart J., Ubbels G., Black S., Hara K., Kirschner M. A reinvestigation of the role of the grey crescent in axis formation in xenopus laevis. Nature. 1981 Aug 6;292(5823):511–516. doi: 10.1038/292511a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Huchon D., Crozet N., Cantenot N., Ozon R. Germinal vesicle breakdown in the Xenopus laevis oocyte: description of a transient microtubular structure. Reprod Nutr Dev. 1981;21(1):135–148. doi: 10.1051/rnd:19810112. [DOI] [PubMed] [Google Scholar]
  8. Kauffman S., Wille J. J. The mitotic oscillator in Physarum polycephalum. J Theor Biol. 1975 Nov;55(1):47–93. doi: 10.1016/s0022-5193(75)80108-1. [DOI] [PubMed] [Google Scholar]
  9. Kishimoto T., Kanatani H. Cytoplasmic factor responsible for germinal vesicle breakdown and meiotic maturation in starfish oocyte. Nature. 1976 Mar 25;260(5549):321–322. doi: 10.1038/260321a0. [DOI] [PubMed] [Google Scholar]
  10. Maller J. L., Krebs E. G. Regulation of oocyte maturation. Curr Top Cell Regul. 1980;16:271–311. doi: 10.1016/b978-0-12-152816-4.50012-1. [DOI] [PubMed] [Google Scholar]
  11. Maller J., Wu M., Gerhart J. C. Changes in protein phosphorylation accompanying maturation of Xenopus laevis oocytes. Dev Biol. 1977 Jul 15;58(2):295–312. doi: 10.1016/0012-1606(77)90093-8. [DOI] [PubMed] [Google Scholar]
  12. Masui Y., Clarke H. J. Oocyte maturation. Int Rev Cytol. 1979;57:185–282. doi: 10.1016/s0074-7696(08)61464-3. [DOI] [PubMed] [Google Scholar]
  13. Masui Y., Markert C. L. Cytoplasmic control of nuclear behavior during meiotic maturation of frog oocytes. J Exp Zool. 1971 Jun;177(2):129–145. doi: 10.1002/jez.1401770202. [DOI] [PubMed] [Google Scholar]
  14. Miake-Lye R., Newport J., Kirschner M. Maturation-promoting factor induces nuclear envelope breakdown in cycloheximide-arrested embryos of Xenopus laevis. J Cell Biol. 1983 Jul;97(1):81–91. doi: 10.1083/jcb.97.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nelkin B., Nichols C., Vogelstein B. Protein factor(s) from mitotic CHO cells induce meiotic maturation in Xenopus laevis oocytes. FEBS Lett. 1980 Jan 14;109(2):233–238. doi: 10.1016/0014-5793(80)81094-5. [DOI] [PubMed] [Google Scholar]
  16. Newport J., Kirschner M. A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982 Oct;30(3):675–686. doi: 10.1016/0092-8674(82)90272-0. [DOI] [PubMed] [Google Scholar]
  17. Reynhout J. K., Smith L. D. Studies on the appearance and nature of a maturation-inducing factor in the cytoplasm of amphibian oocytes exposed to progesterone. Dev Biol. 1974 Jun;38(2):394–400. doi: 10.1016/0012-1606(74)90016-5. [DOI] [PubMed] [Google Scholar]
  18. Schuetz A. W., Samson D. Nuclear requirement of post-maturational cortical differentiation of amphibian oocytes: effects of cycloheximide. J Exp Zool. 1979 Nov;210(2):307–319. doi: 10.1002/jez.1402100214. [DOI] [PubMed] [Google Scholar]
  19. Sunkara P. S., Wright D. A., Rao P. N. Mitotic factors from mammalian cells induce germinal vesicle breakdown and chromosome condensation in amphibian oocytes. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2799–2802. doi: 10.1073/pnas.76.6.2799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wasserman W. J., Masui Y. A cytoplasmic factor promoting oocyte maturation: its extraction and preliminary characterization. Science. 1976 Mar 26;191(4233):1266–1268. doi: 10.1126/science.1083070. [DOI] [PubMed] [Google Scholar]
  21. Wasserman W. J., Masui Y. Effects of cyclohexamide on a cytoplasmic factor initiating meiotic naturation in Xenopus oocytes. Exp Cell Res. 1975 Mar 15;91(2):381–388. doi: 10.1016/0014-4827(75)90118-4. [DOI] [PubMed] [Google Scholar]
  22. Wasserman W. J., Smith L. D. The cyclic behavior of a cytoplasmic factor controlling nuclear membrane breakdown. J Cell Biol. 1978 Jul;78(1):R15–R22. doi: 10.1083/jcb.78.1.r15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weintraub H., Buscaglia M., Ferrez M., Weiller S., Boulet A., Fabre F., Baulieu E. E. Mise en évidence d'une activité "MPF" chez Saccharomyces cerevisiae. C R Seances Acad Sci III. 1982 Dec 20;295(13):787–790. [PubMed] [Google Scholar]
  24. Wu M., Gerhart J. C. Partial purification and characterization of the maturation-promoting factor from eggs of Xenopus laevis. Dev Biol. 1980 Oct;79(2):465–477. doi: 10.1016/0012-1606(80)90131-1. [DOI] [PubMed] [Google Scholar]
  25. Ziegler D., Masui Y. Control of chromosome behavior in amphibian oocytes. II. The effect of inhibitors of RNA and protein synthesis on the induction of chromosome condensation in transplanted brain nuclei by oocyte cytoplasm. J Cell Biol. 1976 Mar;68(3):620–628. doi: 10.1083/jcb.68.3.620. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES