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http://dx.doi.org/10.5713/ajas.2004.1069

MAP Kinase is Activated dring the Maturation of Porcine Oocytes  

Chung, Ki-Hwa (Department of Animal Resources Technology, Jinju National University)
Kim, Chul-Wook (Department of Animal Resources Technology, Jinju National University)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.17, no.8, 2004 , pp. 1069-1075 More about this Journal
Abstract
In an attempt to evaluate the function of MAP kinase in porcine oocytes and to develop a method of the assessment of its activity, myelin basic protein (MBP) was used as a substrate to detect the MAP kinase activity of porcine oocytes which had undergone maturation in vitro. The existence of MAP kinase and MAP kinase kinase (MAPKK) was verified in immature porcine germinal vesicle (GV) oocytes at 0 h culture via Western blotting. Porcine oocytes exhibited a low level of MAP kinase activity during the first 20 h of culture, which increased at 25 h, during which time a breakdown in the nuclear membrane occurred. Significantly higher increases (p<0.05) of MAP kinase activity were detected at 30 h of culture. Using the gel phosphorylation method, MBP was phosphorylated at two positions corresponding to mammalian MAP kinase-extracellular signal-regulated kinase (ERK 1) (44 kDa) and ERK 2 (42 kDa). The absolute levels of those proteins did not increase during 40 h of culture, suggesting that the detected increase in MAP kinase activity was the result of phosphorylation rather than changes in the total amount of protein. MAPKK and MAP kinase were dephosphorylated in first-stage (MI) meiotic oocytes by the addition of cycloheximide, a protein synthesis inhibitor. These results of this study indicate that the MAP kinase cascade does exists in porcine oocytes and that its activation leads to oocyte maturation.
Keywords
Porcine Oocytes; Maturation; Western Blotting; MAP Kinase; Phosphorylation;
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