Christmann L, Jung T, Moor R M
Department of Development and Signalling, AFRC Babraham Institute, Cambridge, England.
Mol Reprod Dev. 1994 May;38(1):85-90. doi: 10.1002/mrd.1080380114.
Growing pig oocytes (< or = 90 microns in diameter) are unable to resume meiosis in vitro. The objective of our present experiments has been to identify the reasons for meiotic competence in these cells. By comparing histone H1 kinase activity in growing and fully grown oocytes we demonstrate that incompetence is associated with an inability to activate H1 kinase in growing oocytes. Immunoblotting was used to determine whether this kinase inactivity resulted from a lack of either p34cdc2 protein or B-type cyclin. The results established that each of these cell cycle molecules are present in comparable amounts in both growing and fully grown oocytes. In the third series of studies experiments were carried out in an attempt to induce p34cdc2 activation during growth. Treatment with okadaic acid, an inhibitor of phosphatase 1 and 2A known to stimulate and accelerate the transition into M-phase of the meiotic cycle in a number of different species, was able to induce p34cdc2 kinase activity and facilitated the G2- to M-phase in growing oocytes. We conclude that although growing oocytes in pigs have sufficient key cell cycle components for the G2 to M transition, they remain incapable of converting these components to active maturation-promoting factor (MPF) until growth is virtually completed. Inhibition of phosphatase 1 or 2A induces the formation of active MPF during growth by an as yet unidentified pathway.
生长中的猪卵母细胞(直径≤90微米)在体外无法恢复减数分裂。我们目前实验的目的是确定这些细胞减数分裂能力的原因。通过比较生长中的和完全成熟的卵母细胞中的组蛋白H1激酶活性,我们证明了生长中的卵母细胞减数分裂无能与无法激活H1激酶有关。使用免疫印迹法来确定这种激酶无活性是否是由于缺乏p34cdc2蛋白或B型细胞周期蛋白。结果表明,这两种细胞周期分子在生长中的和完全成熟的卵母细胞中的含量相当。在第三系列研究中,进行了实验以试图在生长过程中诱导p34cdc2激活。用冈田酸处理,已知它是磷酸酶1和2A的抑制剂,能在许多不同物种中刺激并加速减数分裂周期向M期的转变,它能够诱导生长中的卵母细胞中的p34cdc2激酶活性,并促进其从G2期向M期转变。我们得出结论,尽管猪的生长中的卵母细胞具有足够的关键细胞周期成分用于从G2期向M期转变,但在生长几乎完成之前,它们仍然无法将这些成分转化为活性成熟促进因子(MPF)。磷酸酶1或2A的抑制通过一条尚未明确的途径在生长过程中诱导活性MPF的形成。