Wickramasinghe D, Ebert K M, Albertini D F
Tufts University Health Science Schools, Department of Anatomy and Cellular Biology, School of Medicine, Boston, Massachusetts 02111.
Dev Biol. 1991 Jan;143(1):162-72. doi: 10.1016/0012-1606(91)90063-9.
To determine whether the acquisition of meiotic competence during the growth phase of oogenesis is associated with the appearance of M-phase characteristics, oocytes obtained from 13- to 30-day-old mice were evaluated by fluorescence microscopy with respect to chromatin and microtubule organization , in vitro maturation ability, and the distribution of M-phase phosphoproteins. Meiotically incompetent oocytes were distinguished from their competent counterparts in displaying elaborate interphase-like arrays of cytoplasmic microtubules and dispersed germinal vesicle chromatin. Meiotically competent oocytes were larger in size, exhibited condensation of chromatin around the nucleolus, and displayed a progressive diminution of cytoplasmic microtubules in conjunction with the appearance of multiple microtubule organizing centers. After 24 hr in culture, medium- to large-sized oocytes exhibiting perinucleolar chromatin condensation resume meiosis whereas smaller meiotically incompetent oocytes retain GVs with diffuse chromatin. Moreover, indirect immunofluorescence studies using the M-phase phosphoprotein specific monoclonal antibody MPM-2 indicate that the appearance of reactive cytoplasmic foci is directly correlated with nuclear changes characteristic of meiotically competent oocytes. Thus, the earliest transition to a meiotically competent state during oocyte growth in the immature mouse ovary is characterized by stage-specific and coordinated modifications of nuclear and cytoplasmic components.
为了确定在卵子发生的生长阶段减数分裂能力的获得是否与M期特征的出现相关,对从13至30日龄小鼠获得的卵母细胞进行了荧光显微镜评估,观察其染色质和微管组织、体外成熟能力以及M期磷酸化蛋白的分布。减数分裂无能力的卵母细胞与有能力的卵母细胞的区别在于,前者显示出精细的类似间期的细胞质微管阵列和分散的生发泡染色质。减数分裂有能力的卵母细胞体积更大,在核仁周围表现出染色质凝聚,并随着多个微管组织中心的出现,细胞质微管逐渐减少。培养24小时后,表现出核仁周围染色质凝聚的中大型卵母细胞恢复减数分裂,而较小的减数分裂无能力的卵母细胞则保留具有弥散染色质的生发泡。此外,使用M期磷酸化蛋白特异性单克隆抗体MPM-2进行的间接免疫荧光研究表明,反应性细胞质灶的出现与减数分裂有能力的卵母细胞的核变化直接相关。因此,在未成熟小鼠卵巢中卵母细胞生长过程中最早向减数分裂有能力状态的转变,其特征是核和细胞质成分的阶段特异性和协调性修饰。