Centro de Estudios en Ginecología y Reproducción (CEGyR), Buenos Aires, Argentina.
Mol Hum Reprod. 2011 Jun;17(6):392-8. doi: 10.1093/molehr/gar009. Epub 2011 Feb 5.
The oocyte's meiotic spindle is a dynamic structure that relies on microtubule organization and regulation by centrosomes. Disorganization of centrosomal proteins, including the nuclear mitotic apparatus (NuMA) protein and the molecular motor complex dynein/dynactin, can lead to chromosomal instability and developmental abnormalities. The present study reports the distribution and function of these proteins in human oocytes, zygotes and early embryos. A total of 239 oocytes, 90 zygotes and discarded embryos were fixed and analyzed with confocal microscopy for NuMA and dynactin distribution together with microtubules and chromatin. Microtubule-associated dynein-dependent transport functions were explored by inhibiting phosphatase and ATPase activity with sodium-orthovanadate (SOV). At germinal vesicle (GV) stages, NuMA was dispersed across the nucleoplasm. After GV breaks down, NuMA became cytoplasmic before localizing at the spindle poles in metaphase I and II oocytes. Aberrant NuMA localization patterns were found during oocyte in vitro maturation. After fertilization, normal and abnormal pronuclear stage zygotes and embryos displayed translocation of NuMA to interphase nuclei. SOV treatment for up to 2 h induced lower maturation rates with chromosomal scattering and ectopic localization of NuMA. Accurate distribution of NuMA is important for oocyte maturation, zygote and embryo development in humans. Proper assembly of NuMA is likely necessary for bipolar spindle organization and human oocyte developmental competence.
卵母细胞的减数分裂纺锤体是一种依赖于微管组织和中心体调节的动态结构。中心体蛋白的解聚,包括核有丝分裂装置(NuMA)蛋白和分子马达复合物 dynein/dynactin,可导致染色体不稳定和发育异常。本研究报告了这些蛋白质在人类卵母细胞、受精卵和早期胚胎中的分布和功能。共固定了 239 个卵母细胞、90 个受精卵和废弃胚胎,并通过共聚焦显微镜分析它们的 NuMA 和 dynactin 与微管和染色质的分布。通过用偏钒酸钠(SOV)抑制磷酸酶和 ATP 酶活性来探索微管相关的 dynein 依赖性运输功能。在生发泡(GV)阶段,NuMA 弥散在核质中。GV 破裂后,NuMA 先在细胞质中,然后在减数分裂 I 和 II 期卵母细胞中定位在纺锤体两极。在体外成熟的卵母细胞中发现了异常的 NuMA 定位模式。受精后,正常和异常的原核期受精卵和胚胎显示 NuMA 向间期核的易位。SOV 处理长达 2 小时会导致染色体分散和 NuMA 异位定位,从而降低成熟率。NuMA 的准确分布对于人类卵母细胞成熟、受精卵和胚胎发育是重要的。NuMA 的正确组装可能对于双极纺锤体组织和人类卵母细胞发育能力是必要的。