秋水仙碱化学去核对山羊卵丘细胞核移植胚胎体外发育的影响

研究旨在探讨秋水仙碱化学去核在山羊卵母细胞去核中的应用及其对卵丘细胞核移植胚胎体外发育的影响。结果表明:用秋水仙碱处理山羊体外成熟19h的卵母细胞,可使其核区形成胞质突起,以此为指示进行显微操作去核率可达100%,且用0.8μg/mL秋水仙碱处理30min的山羊卵母细胞突起率可达91.27%,显著高于对照组及0.4μg/mL处理组(P<0.05),与1.6μg/mL处理组的突起率差异不显著(P>0.05)。通过比较盲吸去核法与秋水仙碱化学去核法构建的山羊卵丘细胞核移植胚胎的体外发育能力发现,秋水仙碱化学去核法构建的重构胚体外发育能力较高,桑椹胚发育率显著高于盲吸去核法(P<0.05)。

Effect of Colchicine Assisted-enucleation on in vitro Developmental Potential of Goat Cumulus Cell Nuclear Transferred Embryos

The application of colchicine(a kind of microtubule destabilizing drug) used in the chemical enucleation of goat oocytes was studied in the research and we also analysed its effect on the development rate of goat cumulus cell nuclear transferred embryos.Cytoplasmic protrusions were found in the goat oocytes in vitro matured for 19 h after treatment with colchicine, and the chromatin was in the protrusions as confirmed by Hoechst staining.The enucleation rate would reach 100% when enucleating the protrusions, and we observed that the protrusion rate of goat oocytes was 91.27% when they were treated with 0.8 μg/mL colchicine for 30 min, and it was significantly higher than that of the control group and that of the 0.4 μg/mL colchicine group(P < 0.05), the rate was similar to that of the 1.6 μg/mL colchicine group(P > 0.05).What′s more, we found that the morula rate of cloned embryos reconstrusted by colchicine assisted-enucleation method was significantly higher than that of embryos reconstrusted by blind enucleation method(P < 0.05).