离心对小鼠卵母细胞纺锤体定位和早期发育潜力的影响

Effects of Centrifugation of Mouse Oocytes on the Localization of Meiotic Spindle and the Early Developmental Potential

为了评价离心对卵母细胞纺锤体定位和早期发育潜力的影响,以小鼠MII期卵母细胞经过1000、3 000和5 000 r/min离心10 min,通过免疫荧光细胞化学技术检测纺锤体定位的变化,并通过孤雌激活和体外受精分析离心处理后卵母细胞的发育潜力。结果显示离心后的卵母细胞纺锤体构像正常,定位于皮层边缘,且更靠近第一极体,离心处理的卵孤雌激活后纺锤体动态和减数分裂进程与对照组一致,并保持了正常的早期胚胎发育潜力。这些结果提示卵母细胞的离心处理可作为小鼠核移植中去核的前处理,从而改善在没有霍夫曼镜头情况下盲吸去核的效率。

In order to evaluate the effects of centrifugation on the location of the meiotic spindle and early developmental potential of mouse oocytes, the metaphase-Ⅱ mouse oocytes were centrifuged at 1 000, 3 000 and 5 000 r/min for 10 minutes. The location of the meiotic spindles of centrifuged oocytes was detected by confocal microscopy, and their early developmental potential was analyzed after parthenogenetic activation and in vitro fertilization （IVF）. The results displayed that the meiotic spindles of the centrifuged oocytes showed the normal configuration, and located at the edge of cortex, adjoining the PB 1. The spindle dynamics and the meiotic progress of parthenogenetic activated oocytes treated with centrifugation were consistent with control, and centrifuged oocytes kept the normal early developmental potential after parthenogenetic activation or IVF. The present results imply that the centrifugation-pretreatment of mouse oocytes before enucleation operation may improve the efficiency of enucleation without Hoffmann objective during nuclear transfer.