荧光染色及紫外照射(UV)对黄牛孤雌和克隆胚胎早期发育的影响

Effects of fluorescence staining and ultraviolet(UV) irradiationon the development of parthenogenic and somatic nuclear transfer embryos in bovine

探讨荧光染色及紫外照射对牛孤雌和克隆胚胎体外发育的影响。结果发现,1/4倍UV照射20、30、40 s的卵母细胞孤雌激活后囊胚发育率都极显著低于UV照射0 s和10 s组;以1、1/4、1/8、1/32倍的UV强度照射黄牛卵母细胞20 s,其中1/4倍UV照射组的分裂率与1/8、1/32倍UV照射组无显著差异,但囊胚率间差异显著。以1倍UV强度照射组的分裂率与囊胚率均显著低于1/8、1/32倍UV强度照射组;观察培养24 h后原核形成情况,发现1倍UV强度照射组原核形成也显著滞后于其它处理组;比较1、1/8倍UV强度照射引导去核胞质构建的重组胚体外早期发育情况,发现1倍UV强度照射引导去核胞质重组胚的卵裂率及囊胚率极显著低于1/8倍UV照射强度引导去核胞质组。以上结果表明:1/4倍UV强度照射黄牛卵母细胞时间大于20 s,显著降低其孤雌激活后的卵裂和体外发育潜力;1、1/4倍UV照射强度可阻滞原核形成,降低UV照射强度可提高胚胎孤雌发育率;1/8倍UV照射强度可用于引导去核操作。

Effects of fluorescence staining and ultraviolet irradiation on development of parthenogenic and somatic nuclear transfer embryos of bovine were studied. The cleavage and blastocyst development rates of parthenogenic embryos derived from oocytes irradiated with 1/4 × UV intensity for 20 s and 40 s were significantly lower than those of oocytes irradiated for 0 s and 10 s, indicating irradiated by 1/4 ×UV intensity longer than 20 s can significantly decrease the cleavage and potential development of parthenogenic embryos ; Irradiation of oocytes with 1 ×, 1/4 × UV intensity resulted in a significant decrease in their blastocyst rates or pronuclear formation rates compared with 1/8×, 1/32XUV intensity, indicating that oocytes irradiated by 1×, 1/4× UV intensity could delay the pronuclear formation and decrease the UV irradiation intensity could increase the development of parthenogenic embryos; In cloning research, the cleavage and blastocyst rates of reconstructed embryos derived from cytoplasts irradiated during enucleated by 1 × UV intensity were significantly lower than that of ooeytes suffered from 1/8 ×, UV intensity irradiation, indicating that 1/8 × UV intensity could be emplored for oocyte enucleation in nuclear transfer.