Gordo A C, He C L, Smith S, Fissore R A
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Mol Reprod Dev. 2001 May;59(1):106-14. doi: 10.1002/mrd.1012.
Mammalian oocytes are arrested at the G2/M transition of the first meiotic division from which, after reaching full size and subsequent to an LH surge, they undergo final maturation. Oocyte maturation, which involves germinal vesicle breakdown, progression through metaphase I (MI), and arrest at MII, is triggered and regulated by the coordinated action of two kinases, maturation promoting factor (MPF) and mitogen activated protein kinase (MAPK). The importance of the role of MPF in mammalian oocyte maturation is well established, while the role of MAPK, although well understood in mouse oocytes, has not been fully elucidated in oocytes of large domestic species, especially bovine oocytes. Here we show that injection of MKP-1 mRNA, which encodes a dual specificity MAPK phosphatase, into germinal vesicle stage bovine oocytes prevents the activation of MAPK during maturation. Despite the lack of MAPK activity, MKP-1-injected oocytes resume and progress through meiosis, although they are unable to arrest at MII stage and, by 22-26-hour post-maturation, exhibit decondensed pronucleus-like chromatin, a clear sign of parthenogenetic activation. MKP-1-injected bovine oocytes exhibit normal activation of MPF activity; however, by 18-hour post-maturation, MPF activity starts to decline and by 22-26 hr MPF activity is absent. MKP-1-injected oocytes also show disorganized MII spindles with poorly aligned chromosomes. In summary, our results demonstrate that in bovine oocytes MAPK activity is required for MII arrest, maintenance of MPF activity, and spindle organization.
哺乳动物卵母细胞停滞于第一次减数分裂的G2/M转换期,在达到完全大小并经历促黄体生成素高峰后,它们会经历最终成熟。卵母细胞成熟包括生发泡破裂、通过减数分裂中期I(MI)以及停滞于减数分裂中期II(MII),由两种激酶——成熟促进因子(MPF)和丝裂原活化蛋白激酶(MAPK)的协同作用触发和调节。MPF在哺乳动物卵母细胞成熟中的作用已得到充分证实,而MAPK的作用虽然在小鼠卵母细胞中已被充分了解,但在大型家养动物的卵母细胞中,尤其是牛卵母细胞中尚未完全阐明。在此我们表明,将编码双特异性MAPK磷酸酶的MKP-1 mRNA注射到生发泡期牛卵母细胞中可防止成熟过程中MAPK的激活。尽管缺乏MAPK活性,但注射MKP-1的卵母细胞仍能恢复并通过减数分裂,不过它们无法停滞于MII期,并且在成熟后22 - 26小时,呈现出解聚的原核样染色质,这是孤雌激活的明显迹象。注射MKP-1的牛卵母细胞表现出MPF活性的正常激活;然而,在成熟后18小时,MPF活性开始下降,到22 - 26小时MPF活性消失。注射MKP-1的卵母细胞还显示出MII纺锤体紊乱,染色体排列不佳。总之,我们的结果表明,在牛卵母细胞中,MAPK活性对于MII停滞、MPF活性的维持以及纺锤体组织是必需的。