Journal of Assisted Reproduction and Genetics

, Volume 27, Issue 4, pp 169–182 | Cite as

Dynamics of protein phosphorylation during meiotic maturation

  • Lynda K. McGinnisEmail author
  • David F. Albertini
Gamete Biology



To ask whether distinct kinase signaling pathways mediate cytoplasmic or nuclear maturation of mouse oocytes and if in vitro maturation influences the distribution and timing of these phosphorylation events.


Mouse cumulus oocyte complexes (COCs) were matured under conditions known to influence oocyte quality (basal or supplemented media) and assayed with epitope specific antibodies that would distinguish between Cdk1 or tyrosine kinase targets at 0, 2, 4, 8, and 16 hrs. Semi-quantitative image analysis was used to assess the topographical patterns of protein phosphorylation during in vitro maturation. In vitro fertization and embryo culture were used to examine the effects of culture conditions on developmental potential.


Protein tyrosine phosphorylation increased during meiotic progression from methaphase-I to metaphase-II. Levels were significantly higher in the oocyte cortex. Levels of cortical staining are enhanced in oocytes matured in supplemented media that displayed higher developmental competence. In contrast, bulk substrates for Cdk1 kinase localize to the meiotic spindle while cytoplasmic levels of kinase activity increase throughout meiotic progression; culture media had no measurable effect. Ablation of the tyrosine kinase Fyn significantly reduced cortical levels of tyrosine phosphorylation.


The findings indicate that distinct signaling pathways mediate nuclear and cytoplasmic maturation during in vitro maturation in a fashion consistent with a role for tyrosine kinases in cortical maturation and oocyte quality.


Oocyte maturation Phosphorylation Fertilization Cytoplasmic maturation 



The authors would like to thank Dr. William H. Kinsey for providing the Fyn knock-out mice from his breeding colony. Supported by NICHD 42076, The Hall Family Foundation and the ESHE fund.


  1. 1.
    Swain JE, Smith GD. Reversible phosphorylation and regulation of mammalian oocyte meiotic chromatin remodeling and segregation. Soc Reprod Fertil Suppl. 2007;63:343–58.PubMedGoogle Scholar
  2. 2.
    Besterman B, Schultz RM. Regulation of mouse preimplantation development: inhibitory effect of genistein, an inhibitor of tyrosine protein phosphorylation, on cleavage of one-cell embryos. J Exp Zool. 1990;256:44–53.CrossRefPubMedGoogle Scholar
  3. 3.
    Harrouk W, Clarke HJ. Mitogen-activated protein (MAP) kinase during the acquisition of meiotic competence by growing oocytes of the mouse. Mol Reprod Dev. 1995;41:29–36.CrossRefPubMedGoogle Scholar
  4. 4.
    Ito J, Yoon S-Y, Lee B, Vanderheyden V, Vermassen E, Wojcikiewicz R, et al. Inositol 1, 4, 5-trisphosphate receptor 1, a widespread CA2+ channel, is a novel substrate of polo-like kinase 1 in eggs. Dev Biol. 2008;320:402–13.CrossRefPubMedGoogle Scholar
  5. 5.
    Lee B, Vermassen E, Yoon S-Y, Vanderheyden V, Ito J, Alfandari D, et al. Phosphorylation of IP3R1 and the regulation of [Ca2+]i responses at fertilization: a role for the MAP kinase pathway. Development. 2006;133:4355–65.CrossRefPubMedGoogle Scholar
  6. 6.
    Ohsugi M, Butz S, Kemler R. Beta-Catenin is a major tyrosine-phosphorylated protein during mouse oocyte maturation and preimplantation development. Dev Dyn. 1999;216:168–76.CrossRefPubMedGoogle Scholar
  7. 7.
    Wang Y, Puscheck EE, Lewis JJ, Trostinskaia AB, Wang F, Rappolee DA. Increases in phosphorylation of SAPK/JNK and p38MAPK correlate negatively with mouse embryo development after culture in different media. Fertil Steril. 2005;83:1144.CrossRefPubMedGoogle Scholar
  8. 8.
    Jurema MW, Nogueira D. In vitro maturation of human oocytes for assisted reproduction. Fertil Steril. 2006;86:1277–91.CrossRefPubMedGoogle Scholar
  9. 9.
    Roberts R, Iatropoulou A, Ciantar D, Stark J, Becker DL, Franks S, et al. Follicle-stimulating hormone affects metaphase I chromosome alignment and increases aneuploidy in mouse oocytes matured in vitro. Biol Reprod. 2005;72:107–18.CrossRefPubMedGoogle Scholar
  10. 10.
    McGinnis LK, Albertini DF, Kinsey WH. Localized activation of Src-family protein kinases in the mouse egg. Dev Biol. 2007;306:241–54.CrossRefPubMedGoogle Scholar
  11. 11.
    Meng L, Luo J, Li C, Kinsey WH. Role of Src homology 2 domain-mediated PTK signaling in mouse zygotic development. Reproduction. 2006;132:413–21.CrossRefPubMedGoogle Scholar
  12. 12.
    Zheng K-G, Meng X-Q, Yang Y, Yu Y-S, Liu D-C, Li Y-L. Requirements of Src family kinase during meiotic maturation in mouse oocyte. Mol Reprod Dev. 2007;74:126–31.CrossRefGoogle Scholar
  13. 13.
    Talmor-Cohen A, Tomashov-Matar R, Tsai WB, Kinsey WH, Shalgi R. Fyn kinase-tubulin interaction during meiosis of rat eggs. Reproduction. 2004;128:387–93.CrossRefPubMedGoogle Scholar
  14. 14.
    McGinnis LK, Kinsey WH, Albertini DF. Functions of Fyn kinase in the completion of meiosis in mouse oocytes. Dev Biol. 2009;327:280–7.CrossRefPubMedGoogle Scholar
  15. 15.
    Newhall KJ, Criniti AR, Cheah CS, Smith KC, Kafer KE, Burkart AD, et al. Dynamic anchoring of PKA is essential during oocyte maturation. Curr Biol. 2006;16:321–7.CrossRefPubMedGoogle Scholar
  16. 16.
    Ben-Yosef D, Talmor A, Shwartz L, Granot Y, Shalgi R. Tyrosyl-phosphorylated proteins are involved in regulation of meiosis in the rat egg. Mol Reprod Dev. 1998;149:176–85.CrossRefGoogle Scholar
  17. 17.
    Summers MC, McGinnis LK, Lawitts JA, Biggers JD. Mouse embryo development following IVF in media containing either L-glutamine or glycyl-L-glutamine. Hum Reprod. 2005;20:1364–71.CrossRefPubMedGoogle Scholar
  18. 18.
    Schroeder AC, Schultz RM, Kopf GS, Taylor FR, Becker RB, Eppig JJ. Fetuin inhibits zona pellucida hardening and conversion of ZP2 to ZP2f during spontaneous mouse oocyte maturation in vitro in the absence of serum. Biol Reprod. 1990;43:891–7.CrossRefPubMedGoogle Scholar
  19. 19.
    Spiro RG. Studies on fetuin, a glycoprotein of fetal serum. I. Isolation, chemical composition, and physiochemical properties. J Biol Chem. 1960;235:2860–9.PubMedGoogle Scholar
  20. 20.
    Summers MC, McGinnis LK, Lawitts JA, Raffin M, Biggers JD. IVF of mouse ova in a simplex optimized medium supplemented with amino acids. Hum Reprod. 2000;15:1791–801.CrossRefPubMedGoogle Scholar
  21. 21.
    Messinger SM, Albertini DF. Centrosome and microtubule dymanics during meiotic progression in the mouse oocyte. J Cell Sci. 1991;100:289–98.PubMedGoogle Scholar
  22. 22.
    Pelech S, Jelinkova L, Susor A, Zhang H, Shi X, Pavlok A, et al. Antibody microarray analyses of signal transduction protein expression and phosphorylation during porcine oocyte maturation. J Proteome Res. 2008;7:2860–71.CrossRefPubMedGoogle Scholar
  23. 23.
    Luo J, McGinnis LK, Kinsey WH. Fyn kinase activity is required for normal organization and functional polarity of the mouse oocyte cortex. Mol Reprod Dev. 2009;76:819–31.CrossRefPubMedGoogle Scholar
  24. 24.
    Eppig JJ, Hosoe M, O’Brien MJ, Pendola FM, Requena A, Watanabe S. Conditions that affect acquisition of developmental competence by mouse oocytes in vitro: FSH, insulin, glucose and ascorbic acid. Mol Cell Endocrinol. 2000;163:109.CrossRefPubMedGoogle Scholar
  25. 25.
    Vandre DD, Davis FM, Rao PN, Borisy GG. Phosphoproteins are components of mitotic microtubule organizing centers. Proc Natl Acad Sci U S A. 1984;81:4439–43.CrossRefPubMedGoogle Scholar
  26. 26.
    Davis FM, Tsao TY, Fowler SK, Rao PN. Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci U S A. 1983;80:2926–30.CrossRefPubMedGoogle Scholar
  27. 27.
    Wickramasinghe D, Ebert KM, Albertini DF. Meiotic competence acquisition is associated with the appearance of M-phase characteristics in growing mouse oocytes. Dev Biol. 1991;143:162–72.CrossRefPubMedGoogle Scholar
  28. 28.
    Reis A, Madgwick S, Chang H-Y, Nabti I, Levasseur M, Jones KT. Prometaphase APCcdh1 activity prevents non-disjunction in mammalian oocytes. Nat Cell Biol. 2007;9:1192.CrossRefPubMedGoogle Scholar
  29. 29.
    Kapus A, Di Ciano C, Sun J, Zhan X, Kim L, Wong T, et al. Cell volume-dependent phosphorylation of proteins of the cortical cytoskeleton and cell-cell contact sites. THE ROLE OF Fyn AND FER KINASES. J Biol Chem. 2000;275:32289–98.CrossRefPubMedGoogle Scholar
  30. 30.
    Thomas SM, Soriano P, Imamoto A. Specific and redundant roles of Src and Fyn in organizing the cytoskeleton. Nature. 1995;376:267.CrossRefPubMedGoogle Scholar
  31. 31.
    Tournaviti S, Hannemann S, Terjung S, Kitzing TM, Stegmayer C, Ritzerfeld J, et al. SH4-domain-induced plasma membrane dynamization promotes bleb-associated cell motility. J Cell Sci. 2007;120:3820–9.CrossRefPubMedGoogle Scholar
  32. 32.
    Maekawa M, Toyama Y, Yasuda M, Yagi T, Yuasa S. Fyn tyrosine kinase in sertoli cells is involved in mouse spermatogenesis. Biol Reprod. 2002;66:211–21.CrossRefPubMedGoogle Scholar
  33. 33.
    Li L, Baibakov B, Dean J. A subcortical maternal complex essential for preimplantation mouse embryogenesis. Dev Cell. 2008;15:416–25.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Anatomy & Cell BiologyUniversity of Kansas Medical CenterKansas CityUSA
  2. 2.Department of Integrative PhysiologyUniversity of Kansas Medical CenterKansas CityUSA

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