Sobajima T, Aoki F, Kohmoto K
Department of Animal Breeding, Faculty of Agriculture, University of Tokyo, Japan.
J Reprod Fertil. 1993 Mar;97(2):389-94. doi: 10.1530/jrf.0.0970389.
Mitogen-activated protein kinase (MAP kinase) plays a role in the cascade of protein kinase activation in cultured cells. To investigate the involvement of MAP kinase in meiotic maturation, we measured MAP kinase activity, using myelin basic protein as a substrate, with histone H1 kinase activity, in mouse oocytes. MAP kinase activity was low 1 h after isolation from follicles (when oocytes lost their germinal vesicle), increased abruptly at 2 h, and remained high until the second metaphase (13 h after isolation from follicles). Histone H1 kinase activity increased gradually from 2 to 7 h after isolation. When immature oocytes were treated with puromycin, MAP kinase activity did not increase after isolation from follicles. In the presence of 3-isobutyl-1-methylxanthine, the treatment of immature oocytes with okadaic acid, a specific inhibitor of protein phosphatase 1 and 2A, induced germinal vesicle breakdown and activation of MAP kinase. These results suggest that MAP kinase is involved in the regulation of meiotic maturation, and that the activation of MAP kinase requires protein synthesis and is inhibited by the protein phosphatase during meiotic maturation in mouse oocytes.
丝裂原活化蛋白激酶(MAP激酶)在培养细胞中的蛋白激酶激活级联反应中发挥作用。为了研究MAP激酶在减数分裂成熟过程中的作用,我们以髓鞘碱性蛋白为底物,在小鼠卵母细胞中测量了MAP激酶活性,并同时测量了组蛋白H1激酶活性。从卵泡中分离1小时后(此时卵母细胞失去生发泡),MAP激酶活性较低,2小时时突然升高,并一直保持高水平直至第二次中期(从卵泡中分离13小时后)。组蛋白H1激酶活性在分离后2至7小时逐渐增加。当用嘌呤霉素处理未成熟卵母细胞时,从卵泡中分离后MAP激酶活性未增加。在存在3-异丁基-1-甲基黄嘌呤的情况下,用蛋白磷酸酶1和2A的特异性抑制剂冈田酸处理未成熟卵母细胞,可诱导生发泡破裂和MAP激酶激活。这些结果表明,MAP激酶参与减数分裂成熟的调控,并且在小鼠卵母细胞减数分裂成熟过程中,MAP激酶的激活需要蛋白质合成,并受到蛋白磷酸酶的抑制。