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Activation of mitogen-activated protein kinase during meiotic maturation in porcine oocytes

Published online by Cambridge University Press:  26 September 2008

Maki Inoue ,
Kunihiko Naito ,
Fugaku Aoki ,
Yutaka Toyoda  and
Eimei Sato
Maki Inoue*
Affiliation:
Institute of Medical Science and Faculty of Agriculture, University of Tokyo, Tokyo, Tokyo, Japan.
Kunihiko Naito
Affiliation:
Institute of Medical Science and Faculty of Agriculture, University of Tokyo, Tokyo, Tokyo, Japan.
Fugaku Aoki
Affiliation:
Institute of Medical Science and Faculty of Agriculture, University of Tokyo, Tokyo, Tokyo, Japan.
Yutaka Toyoda
Affiliation:
Institute of Medical Science and Faculty of Agriculture, University of Tokyo, Tokyo, Tokyo, Japan.
Eimei Sato
Affiliation:
Institute of Medical Science and Faculty of Agriculture, University of Tokyo, Tokyo, Tokyo, Japan.
*
Maki Inoue, Department of Reproductive and Developmental Biology, Institute of Medical Science, University of Tokyo, 4-6-1, Shirokanedai, Minato-ku Tokyo, Japan, 108. Telephone: + 81-3-5449-5332. Fax: + 81-3-5449-5451.
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To investigate the involvement of mitogen-activated protein kinase(MAP kinase) in meiotic maturation of porcine oocytes, we assayed MAP kinase activity using basic protein(MBP) as a substrate. MAP kinase activity was low during the germinal vesicle stage, 0–20 h of culture. An abrupt increase was observed at metaphase I(30 h of culture), and activity remained significantly higher than that at 0 h until 50 h of culture, with a transient slight decrease at the time of first polar body extrusion (40 h). Detection of the kinase activity by an in-gel phosphorylation assay confirmed that the 42 and 44 kDa MAP kinases were significantly activated in 45 h cultured oocytes but not in 0 h oocytes, and just slightly in 20 h oocytes. In immunoblotting, however, the 42 and 44 kDa bands were detected in 0, 20 and 45 h cultured oocytes. Furthermore, the signal strength of the two bands did not change during the period of culture, but shifted up to 45 h, indicating that the activation of MAP kinase depended not on the synthesis but on the phosphorylation of this enzyme. These results suggest that the activation of MAP kinase is involved in the regulation of meiotic maturation of porcine oocytes, and especially in the regulation after germinal vesicle breakdown.

Keywords

In vitro maturationMAP kinaseMeiosisOocytePig
Type
Article
Information
Zygote , Volume 3 , Issue 3 , August 1995 , pp. 265 - 271
Copyright
Copyright © Cambridge University Press 1995

References

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