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孵育和激活剂作用时间对山羊核移植胚胎早期发育的影响
李斌,曹陈冲,陈立子,汪存利,赵焕,李东伟,陶勇,章美林,章孝荣
0
(上海交通大学医学院附属第九人民医院,上海 200011; 安徽农业大学动物科技学院,合肥 233036)
摘要:
探讨了电融合前对重构卵继续孵育及激活剂6-DMAP不同作用时间对山羊核移植胚胎体外发育的影响。结果显示,重构卵在成熟液中继续培养后,9~10 h组的融合后卵子死亡率(27.42%)显著低于30 min和90 min组(49.26%和56.3%) (P<0.05), 胚胎卵裂率(68.15%)则显著高于30 min和90 min组(55.6%和54.24%) (P<0.05);重构卵用离子霉素激活后,再用6-DMAP激活0 h、2 h、3 h、4 h、5 h及8~10 h,各组间的胚胎卵裂率均无显著差异(P>0.05),但均显著高于未用6-DMAP组的卵裂率(P<0.05)。本试验结果表明,电融合前,继续成熟培养重构卵有利于降低电融合时的卵子死亡率,提高卵子利用率;联合使用离子霉素和6-DMAP有利于提高激活效果,而6-DMAP的作用时间(2~10 h)不影响核移植胚胎的卵裂。
关键词:  山羊  重构卵  核移植胚胎  化学激活  6-DMAP
DOI:
基金项目:上海高校选拔培养优秀青年教师科研专项基金(zzjdyx12106), 安徽省“十五”重大科技专项(04003005)和“十一五”科技攻关项目(06013051A)共同资助。
Effects of incubation time and activation time of 6-DMAP on the early development of the goat nuclear transplanted embryos
LI Bin,CAO Chenchong,CHEN Lizi,WANG Cunli,ZHAO Huan,LI Dongwei,TAO Yong,ZHANG Meiling,ZHANG Xiaorong
(Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011; School of Animal Sciences and Technology , Anhui Agricultural University, Hefei 230036)
Abstract:
The study was conducted to examine the effects of incubation time before the electrofusion and activation time of 6-DMAP on the development of the goat nuclear transplanted embryos in vitro. The reconstructed oocytes were continually cultured in a maturation medium for 30 min, 90 min and 9-10 h before the electrofusion, respectively. The results showed that the mortality of reconstructed oocytes of 9-10 h group after the electrofusion was significantly lower than those of 30 min and 90 min groups (27.42% vs. 49.26% and 56.3%, P<0.05), while the cleavage rate of 9-10 h group was significantly higher than those of 30 min and 90 min groups (68.15% vs. 55.6% and 54.24%, P<0.05). The nuclear transplanted embryos activated by ionomycin were activated with 6-DMAP for 0 h, 2 h, 3 h, 4 h, 5 h, and 8-10 h respectively. The results showed that the cleavage rate was not significantly different among groups (P>0.05), but it was significantly higher than that of the embryos without 6-DMAP (P<0.05). In conclusion, the continued incubation of reconstructed oocytes before electrofusion may be helpful for lowing the mortality of fusion and increasing the rate of available oocyte; the combination of ionomycin with 6-DMAP could improve the activation efficiency in goat nuclear transplanted embryos, and activation time of 6-DMAP has no significant effects on the cleavage of goat nuclear transplanted embryos.
Key words:  goat  reconstructed ooctye  nuclear transplanted embryo  chemical activation  6-DMAP

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