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Dynamics of protein phosphorylation during meiotic maturation

  • Gamete Biology
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Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.

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Acknowledgements

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.

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Authors and Affiliations

  1. Department of Anatomy & Cell Biology, University of Kansas Medical Center, 3901 Rainbow Blvd., mail stop 3038, Kansas City, KS, 66160, USA

    Lynda K. McGinnis

  2. Department of Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA

    David F. Albertini

Authors
  1. Lynda K. McGinnis
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  2. David F. Albertini
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Corresponding author

Correspondence to Lynda K. McGinnis.

Additional information

Capsule Tracking of distinct kinase targets during meiotic progression of mouse oocytes suggests that oocyte quality is determined by the action of tyrosine kinases on the cortex illustrating a novel aspect of cytoplasmic maturation in mammalian oocytes.

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McGinnis, L.K., Albertini, D.F. Dynamics of protein phosphorylation during meiotic maturation. J Assist Reprod Genet 27, 169–182 (2010). https://doi.org/10.1007/s10815-010-9391-x

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  • DOI: https://doi.org/10.1007/s10815-010-9391-x

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