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Protein Kinase Assays for Measuring MPF and MAPK Activities in Mouse and Rat Oocytes and Early Embryos

  • Jacek Z. KubiakEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 957)

Abstract

Protein phosphorylation plays a pivotal role in cell cycle regulation. MPF (M-phase Promoting Factor) and MAPK (Mitogen-activated protein kinase) are two major kinases driving oocyte maturation and early embryonic divisions. Their activities can be measured experimentally with kinase assays that use specific exogenous substrates. The activities of MPF and MAPK are measured using histone H1 kinase and MBP (Myelin Basic Protein) kinase assays, respectively. Here, we describe detailed procedures for measuring these two activities in mouse and rat oocytes and in early mouse embryos. The assays we describe can be performed using very small amounts of biological material and produce clearly discernible measurements of histone H1 and MBP kinase activities.

Key words

Cell cycle CDK1/cyclin B ERK 1/2 Histone H1 kinase assay MAPK MBP kinase assay Meiosis Mitosis Mouse MPF Oocyte maturation Rat 

Notes

Acknowledgments

I thank Malgorzata Kloc for valuable discussions and critical reading of the manuscript. This work was supported by ARC grants to JZK.

References

  1. 1.
    Kubiak JZ, Ciemerych MA, Hupalowska A, Sikora-Polaczek M, Polanski Z (2008) On the transition from the meiotic to mitotic cell cycle during early mouse development. Int J Dev Biol 52:201–217PubMedCrossRefGoogle Scholar
  2. 2.
    Marteil G, Richard-Parpaillon L, Kubiak JZ (2009) Role of oocyte quality in meiotic maturation and embryonic development. Reprod Biol 9:203–224PubMedCrossRefGoogle Scholar
  3. 3.
    Masui Y, Markert CL (1971) Cytoplasmic control of nuclear behavior during meiotic maturation of frog oocytes. J Exp Zool 177:129–145PubMedCrossRefGoogle Scholar
  4. 4.
    Balakier H, Czolowska R (1977) Cytoplasmic control of nuclear maturation in mouse oocytes. Exp Cell Res 110:466–469PubMedCrossRefGoogle Scholar
  5. 5.
    Gautier J, Norbury C, Lohka M, Nurse P, Maller J (1988) Purified maturation-promoting factor contains the product of a Xenopus homolog of the fission yeast cell cycle control gene cdc2+. Cell 54:433–439PubMedCrossRefGoogle Scholar
  6. 6.
    Gautier J, Minshull J, Lohka M, Glotzer M, Hunt T, Maller JL (1990) Cyclin is a component of maturation-promoting factor from Xenopus. Cell 60:487–494PubMedCrossRefGoogle Scholar
  7. 7.
    Labbé JC, Picard A, Karsenti E, Dorée M (1988) An M-phase-specific protein kinase of Xenopus oocytes: partial purification and possible mechanism of its periodic activation. Dev Biol 127:157–169PubMedCrossRefGoogle Scholar
  8. 8.
    Labbé JC, Picard A, Peaucellier G, Cavadore JC, Nurse P, Doree M (1989) Purification of MPF from starfish: identification as the H1 histone kinase p34cdc2 and a possible mechanism for its periodic activation. Cell 57:253–263PubMedCrossRefGoogle Scholar
  9. 9.
    Arion D, Meijer L (1989) M-phase-specific protein kinase from mitotic sea urchin eggs: cyclic activation depends on protein synthesis and phosphorylation but does not require DNA or RNA synthesis. Exp Cell Res 183:361–375PubMedCrossRefGoogle Scholar
  10. 10.
    Meijer L, Arion D, Golsteyn R, Pines J, Brizuela L, Hunt T, Beach D (1989) Cyclin is a component of the sea urchin egg M-phase specific histone H1 kinase. EMBO J 8:2275–2282PubMedGoogle Scholar
  11. 11.
    Felix MA, Pines J, Hunt T, Karsenti E (1989) A post-ribosomal supernatant from activated Xenopus eggs that displays post-translationally regulated oscillation of its cdc2+ mitotic kinase activity. EMBO J 8:3059–3069PubMedGoogle Scholar
  12. 12.
    Kubiak J, Paldi A, Weber M, Maro B (1991) Genetically identical parthenogenetic mouse embryos produced by inhibition of the first meiotic cleavage with cytochalasin D. Development 111:763–769PubMedGoogle Scholar
  13. 13.
    Weber M, Kubiak JZ, Arlinghaus RB, Pines J, Maro B (1991) c-mos proto-oncogene product is partly degraded after release from meiotic arrest and persists during interphase in mouse zygotes. Dev Biol 148:393–397PubMedCrossRefGoogle Scholar
  14. 14.
    Kubiak JZ, Weber M, Géraud G, Maro B (1992) Cell cycle modification during the transitions between meiotic M-phases in mouse oocytes. J Cell Sci 102:457–467PubMedGoogle Scholar
  15. 15.
    Szöllösi MS, Kubiak JZ, Debey P, de Pennart H, Szöllösi D, Maro B (1993) Inhibition of protein kinases by 6-dimethylaminopurine accelerates the transition to interphase in activated mouse oocytes. J Cell Sci 104:861–872PubMedGoogle Scholar
  16. 16.
    Kubiak JZ, Weber M, de Pennart H, Winston NJ, Maro B (1994) The metaphase II arrest in mouse oocytes is controlled through microtubule-dependent destruction of cyclin B in the presence of CSF. EMBO J 12:3773–3778Google Scholar
  17. 17.
    Verlhac MH, Kubiak JZ, Clarke HJ, Maro B (1994) Microtubule and chromatin behavior follow MAP kinase activity but not MPF activity during meiosis in mouse oocytes. Development 120:1017–1025PubMedGoogle Scholar
  18. 18.
    Ciemerych MA, Tarkowski AK, Kubiak JZ (1998) Autonomous activation of histone H1 kinase, cortical activity and microtubule organization in one- and two-cell mouse embryos. Biol Cell 90:557–564PubMedCrossRefGoogle Scholar
  19. 19.
    Ciemerych MA, Maro B, Kubiak JZ (1999) Control of duration of the first two mitoses in a mouse embryo. Zygote 7:293–300PubMedCrossRefGoogle Scholar
  20. 20.
    Maciejewska Z, Pascal A, Kubiak JZ, Ciemerych MA (in press) Phosphorylated ERK5/BMK1 transiently accumulates within division spindles in mouse oocytes and preimplantation embryos. Folia Histochem Cytobiol 49(3):528–534Google Scholar
  21. 21.
    Skoufias DA, Indorato RL, Lacroix F, Panopoulos A, Margolis RL (2007) Mitosis persists in the absence of Cdk1 activity when proteolysis or protein phosphatase activity is suppressed. J Cell Biol 179:671–685PubMedCrossRefGoogle Scholar
  22. 22.
    Zernicka-Goetz M, Verlhac M-H, Géraud G, Kubiak JZ (1997) Protein phosphatases control MAP kinase activation and microtubule organization during rat oocyte maturation. Eur J Cell Biol 72:30–38PubMedGoogle Scholar

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© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Cell Cycle GroupCNRS, UMR 6290, Institute of Genetics and Development of Rennes (IGDR)RennesFrance
  2. 2.Faculty of MedicineUniversity Rennes 1, UEB, UMS 3480RennesFrance

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