摘要
牛卵母细胞在体外成熟２３～２４ｈ时去核，去核卵用８０Ｖ／ｍｍ４０μｓ两次电脉冲激活（Ⅰ组）或不激活（Ⅱ组），然后将体外受精、发育的８～３２细胞期胚胎的单个卵裂球注入卵周隙，用８０Ｖ／ｍｍ４０μｓ两次电脉冲诱导卵裂球与去核卵母细胞融合。Ⅰ组操作后存活率和融合率（８８．４％和５５．０％）显著低于Ⅱ组（９５．９％和６５．１％，Ｐ＜０．０５）。融合卵体外培养２４ｈ和５～８ｄ后，Ⅰ组核移植胚胎的卵裂率和桑椹／囊胚发育率（６３．０％和１５．２％）与Ⅱ组（５０．５％和７．８％）无显著差异，但Ⅰ组结果均好于Ⅱ组。将来自两组的２５枚桑椹和囊胚移植到１６头同期受体，在已检查过的８头受体中２头受体妊娠，其中１头于妊娠后４个多月流产，１头于妊娠期满后产出一雄性牛犊。研究结果表明：去核卵母细胞的电激活尽管对重组卵的存活与融合不利，但可改善核移植胚的卵裂和发育。本研究在我国首次获得牛细胞核移植的成功，证明用ＩＶＭ卵母细胞和ＩＶＦ胚胎进行完全体外化的牛细胞核移植是可行的。
Bovine oocytes were enucleated after matured in vitro for 23～24h. The enucleated oocytes were electrically activated with two 80V/mm 40us DC pulses(GI) or non-activated(GII).Then a single blastomere from 8～32-cell bovine embryos, which fertilized and developed in vitro, was microinjected into the perivitelline space of the enucleated oocyte.The fusion was induced by two 80/v 40us DC electric pulses, 1 sec interval.The survival rate and fusion rate of GI(88.4%and 55.0%,respectively)were lower than those of GII significantly(95.9%and 65.1%,respectively)(P<0.05).Although there were no significant difference in the cleavage rate(63.0% vs. 50.5%)and the developmental rate of morulae and blastocysts(15.2% vs. 7.8%)between GI and GII after the fused eggs were cultured for 24h or 5～8 days, both rates of GI were higher than GII. Twenty-five morulae and blastocysts were transferred to 16 synchronous recipients. Out of 8recipients diagnosed, 2 were pregnant. Among the two pregnant recipients, one aborted 4months later, another gave birth to a male calf at full-term.In conclusion, although the electric activation of the enucleated has a disadvantageous effect to the survival and fusion of the reconstructed eggs, it improves the cleavage and development of the nuclear transferred embryos. It is the first successful bovine nuclear transfer in China,indicating that it is feasible to transplant bovine nuclear in the whole in vitro procedure using IVM oocytes and IVF embryos.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
1996年第6期495-500,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
广东省"八五"农业科技重点攻关项目
广西教委科学基金
关键词
电激活
体外成熟
体外受精
细胞核移植
牛
Electric activation,In vitro maturation, In vitro fertilization, Nuclear transfer, Bovine
