Cui Long-Bo, Huang Xiu-Ying, Sun Fang-Zhen
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China.
Hum Reprod. 2005 Jun;20(6):1624-31. doi: 10.1093/humrep/deh826. Epub 2005 Mar 10.
[corrected] Transferring a germinal vesicle (GV) from an aged woman's oocyte into ooplasm from a younger woman has been proposed as a possible way to overcome the problem of age-related decline in female fertility. Here we assessed this possibility by determining whether ooplasts derived from young mice could rescue ageing-associated chromosome misalignment in meiosis of oocytes from aged mice.
Three groups of reconstructed oocytes, young GV-young cytoplast (group YY), aged GV-young cytoplast (group AY), and young GV-aged cytoplast (group YA), were created by micromanipulation and electrofusion.
Nuclear transplantation was successful in 89.8-94.4% of GV-ooplast complexes, and maturation rate of the reconstructed oocytes was 93.5-97.9%. Confocal microscopy analysis showed a significantly higher rate (49.2%) of chromosome misalignment in ageing mice than in young mice (16.9%), and 57.1% of oocytes in group AY exhibited chromosome misalignment, while the abnormality rate in groups YY and YA was 16.3 and 16.7% respectively. Calcium imaging showed that the three groups of reconstructed oocytes exhibited a similar pattern of calcium oscillations upon stimulation with bovine sperm extracts. Fertilization rate and developmental capacity to 2-cell embryos were also similar among the three groups of oocytes.
Our findings suggest that: (i) the ooplasm from young mice could not rescue ageing-associated chromosome misalignment in meiosis of GV from aged mice; and (ii) behaviour of chromosome alignment over metaphase spindle is predominantly determined by GV material.
[已修正] 将老年女性卵母细胞的生发泡(GV)转移至年轻女性的卵质中,被认为是克服女性生育力随年龄下降问题的一种可能方法。在此,我们通过确定源自年轻小鼠的卵质体是否能挽救老年小鼠卵母细胞减数分裂中与衰老相关的染色体排列错误,来评估这种可能性。
通过显微操作和电融合创建了三组重构卵母细胞,即年轻GV-年轻细胞质体组(YY组)、老年GV-年轻细胞质体组(AY组)和年轻GV-老年细胞质体组(YA组)。
89.8% - 94.4%的GV-卵质体复合物核移植成功,重构卵母细胞的成熟率为93.5% - 97.9%。共聚焦显微镜分析显示,衰老小鼠中染色体排列错误的发生率(49.2%)显著高于年轻小鼠(16.9%),AY组中57.1%的卵母细胞出现染色体排列错误,而YY组和YA组的异常率分别为16.3%和16.7%。钙成像显示,三组重构卵母细胞在用牛精子提取物刺激时表现出相似的钙振荡模式。三组卵母细胞的受精率和发育至2细胞胚胎的能力也相似。
我们的研究结果表明:(i)年轻小鼠的卵质不能挽救老年小鼠GV减数分裂中与衰老相关的染色体排列错误;(ii)中期纺锤体上染色体排列行为主要由GV物质决定。