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Mitogen-activated protein kinase phosphorylation patterns in pig oocytes and cumulus cells during gonadotrophin-induced resumption of meiosis in vitro

Published online by Cambridge University Press:  01 May 2007

S. Ebeling ,
C. Schuon  and
B. Meinecke
S. Ebeling
Affiliation:
University of Veterinary Medicine, Hannover, Germany
C. Schuon
Affiliation:
University of Veterinary Medicine, Hannover, Germany
B. Meinecke*
Affiliation:
University of Veterinary Medicine, Hannover, Germany
*
All correspondence to: Burkhard Meinecke, Department of Reproductive Biology, University of Veterinary Medicine Hannover, Foundation, Buenteweg 2, D-30559 Hannover, Germany. Tel: +49 511 953 7181. Fax: +49 511 953 7150. e-mail: Burkhard.Meinecke@tiho-hannover.de
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Summary

The present study investigated the phosphorylation pattern of mitogen-activated protein kinase (MAPK) in cumulus–oocyte complexes (COCs) during spontaneous and FSH/LH-induced in vitro maturation (IVM). Both isoforms of MAPK were unphosphorylated in oocytes recovered immediately after liberation from follicles and became phosphorylated following 25 h incubation, corresponding to the time of germinal vesicle breakdown (GVBD). In contrast, MAPK was already phosphorylated in minimal amounts in cumulus cells at the time of liberation from follicles and phosphorylation of MAPK increased after 0.5 h incubation. Supplementation of medium with gonadotrophins intensified phosphorylation at 0.5 h incubation, demonstrating the early and rapid action of FSH/LH on MAPK phosphorylation. Phosphorylation of MAPK in cumulus cells peaked after 21 h of incubation, whereas MAPK was almost completely dephosphorylated at the end of incubation (45 h). During subsequent incubation in the absence of added gonadotrophins, between 5 and 10 h exposure to FSH/LH-supplemented medium was required to induce resumption of meiosis in COCs. Phosphorylation of MAPK in oocytes was prevented by the MEK inhibitor U0126, but the inhibitor reduced phosphorylation of MAPK in cumulus cells only during the first 2 h of IVM. The data support the hypothesis that two different MAPK phosphorylation events occurred following gonadotrophin stimulation, one in cumulus cells and the other in oocytes. In cumulus cells, FSH/LH induced early and rapid U0126-insensitive phosphorylation of MAPK, whereas U0126-susceptible MAPK phosphorylation took place in the oocyte itself around the time of GVBD.

Keywords

Cumulus cellsMAPKMeiotic resumptionPig oocyteU0126
Type
Research Articles
Information
Zygote , Volume 15 , Issue 2 , May 2007 , pp. 139 - 147
Copyright
Copyright © Cambridge University Press 2007

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