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Okadaic acid-sensitive phosphatase is related to MII/G1 transition in mouse oocytes

Published online by Cambridge University Press:  10 February 2011

Naoko Moride
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Akira Kuwahara*
Affiliation:
Department of Obstetrics & Gynecology, University of Tokushima Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Ayako Sutoh
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Yu Tanaka
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Yukari Mukai
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Mizuho Yamashita
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Toshiya Matsuzaki
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Toshiyuki Yasui
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
Minoru Irahara
Affiliation:
Department of Obstetrics and Gynecology, University of Tokushima, Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan.
*
All correspondence to: Akira Kuwahara. Department of Obstetrics & Gynecology, University of Tokushima Institute for Health Biosciences, 3–18–15 Kuramoto Tokushima 770–8503, Japan. Tel: +81 88 633 7177. Fax: +81 88 631 2630. e-mail: [email protected]

Summary

It is reported that okadaic acid (OA)-sensitive phosphatase is related to mitogen-activated protein kinase (MAPK)/p90rsk activation in mammalian oocytes. OA is also involved in the positive feedback loop between M phase-promoting factor (MPF) and cdc25c in Xenopus oocytes during meiotic maturation. However, the effect of phosphatase inhibition by OA on MPF and MAPK activities at the MII/G1 in oocytes remains unknown. The aim of this study is to clarify the relationship between OA-sensitive phosphatase and mitosis MII/G1 transition in mouse oocytes. MII-arrested oocytes were, isolated from mice, inseminated and cultured in TYH medium (control group) or TYH medium supplemented with 2.5 μM of OA (OA group). Histone H1 kinase and myelin basic protein (MBP) kinase activities were measured as indicators of MPF and p42 MAPK activities after insemination. Phosphorylation of cdc25c after insemination was analized in OA and control group by western blotting. Seven hours after insemination a pronucleus (PN) was formed in 84.1% (69/85) of oocytes in the control group. However, no PN was formed in oocytes of the OA group (p < 0.001). Although MPF and MAPK activities in the control group significantly decreased at 3, 4, 5, and 7 h after insemination, these decreases were significantly inhibited by OA addition (p < 0.05). Furthermore, OA addition prevented cdc25c dephosphorylation 7 h after insemination. In conclusion, OA-sensitive phosphatase correlates with inactivation of MPF and MAPK, and with the dephosphorylation of cdc25c at the MII/G1 transition in mouse oocytes.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2011

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