摘要
通过显微操作，将１６～３２细胞期胚胎的单个卵裂球注入去核卵母细胞的卵周隙内，用适当电压（兔１６０Ｖ、牛１２０Ｖ）和脉冲时间的两次电脉冲诱导卵裂球与卵母细胞的融合；融合后的胚胎体外培养观察发育或移植到同步受体。体内成熟的免卵母细胞操作后存活率为９３．７％（１５０１／１６０２），融合率为９４．３％（１２８１／１３５９）。融合卵再用１６０Ｖ４０μｓ电脉冲电激一次，可显著提高其卵裂率（７５．９％比１８．２％，Ｐ＜０．０１）。用体外成熟的兔卵母细胞进行核移植，存活率和融合率分别为７８．５％（９５／１２１）和９５．７％（８８／９２），融合胚卵裂率为５１．４％（３７／７２）。初步表明体外成熟的兔卵母细胞可作为核移植的受体卵。此外，还尝试了用体外受精的牛胚胎和体外成熟的牛卵母细胞进行核移植，操作后存活率和融合率分别为９８．０％（５０／５１）和７５．５％（３７／４９），核移植胚的卵裂率为５４．１％（２０／３７）。
The results of nuclear transfer of oocytes in rabbit and cattle during 1991￣ 1993 were reported in this paper. The single blastomere of 16￣32 cell stage embryos was microinjected into the perivitelline space of enucleated oocyte. The fusion between blas tomere and enucleated oocyte was induced with two DC pulses (rabbit 160V 40μs, cattle 120V 40μs). The reconstructed oocytes were cultured in vitro or transferred to the synchronous recipients to evaluate development ability. Near 94% of rabbit oocytes maturedin vivo survived the micromanipulation (1 501/1 602 ), and the fusion rate of them was 94. 3% (1 281/1 359). Exposing nuclear transfer oocytes to an additional 160V 40μs DC pulse after fusion increased significantly the cleavage rate (75. 9% vs 18. 2%, P<0. 01).Rabbit oocytes matured in vitro resulted in a survival rate of 78. 5 % (95/121 ), a fusion rate of 95. 7% (88/92) and a cleavage rate of 51. 4% (37/72) after micromanipulation,electrofusion and culture in vitro. An attempt also made to transfer nucleus from IVF embryos to enucleated IVM oocytes in cattle. The survival rate, fusion rate and cleavage rate of cattle nuclear transfer oocytes were 98. 0% (50/51), 75. 5% (37/49) and 54. 1 % (20/ 37) respectively.
关键词
兔
牛
卵母细胞
体外成熟
核移植
rabbit
cattle
oocyte
maturation in vivo
maturation in vitro
nuclear transfer
