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刘同明, 俞小牧, 叶玉珍 王忠卫, 周建峰, 吴清江, . 2002: 鱼类体细胞移核胚胎发育影响因素的探讨. 动物学研究, 23(2): 107-112.
引用本文: 刘同明, 俞小牧, 叶玉珍 王忠卫, 周建峰, 吴清江, . 2002: 鱼类体细胞移核胚胎发育影响因素的探讨. 动物学研究, 23(2): 107-112.
LIU Tong-ming, YU Xiao-mu, YE Yu-zhen, WANG Zhong-wei, ZHOU Jian-feng, WU Qing-jiang. 2002. Studies on Factors Affecting the Development of Embryos from Somatic Cell Nuclear Transplantation in the Fish. Zoological Research, 23(2): 107-112.
Citation: LIU Tong-ming, YU Xiao-mu, YE Yu-zhen, WANG Zhong-wei, ZHOU Jian-feng, WU Qing-jiang. 2002. Studies on Factors Affecting the Development of Embryos from Somatic Cell Nuclear Transplantation in the Fish. Zoological Research, 23(2): 107-112.

鱼类体细胞移核胚胎发育影响因素的探讨

Studies on Factors Affecting the Development of Embryos from Somatic Cell Nuclear Transplantation in the Fish

  • 摘要: 将脱膜后的泥鳅(Misgurnus anguillicaudatus)受精卵置于不同孵化液中孵化,结果表明,孵化液1×Holtfreter中常鱼苗和总鱼苗的孵化率最高,分别为54.8%和59.2%。1/2×Holtfreter、1/10×Holtfreter、曝气冷开水和改变孵化液处理,正常鱼苗的孵化率差别不大,分别为48.0%、49.6%、50.0%和48.8%。曝气冷开水畸形率最高,为8.8%。PBS不适合用作孵化液。机械损伤对胚胎的孵化率无明显影响(73.39% vs.75.70%),但对胚胎发育有明显的致畸作用.06% vs.0)。以雌核发育鳙(Aristichthys nobilis)尾鳍培养细胞供体,泥鳅未受精卵作受体,微卫星标记分析表明,移核胚胎的核来源于供体核,受体卵中未除去的核被排斥。供体细胞养代数严重影响移核胚胎的发育率。随着培养代数的增加,核胚胎的发育率逐渐降低。继代核移植可提高移核胚胎的发育率,尤其是第1次体细胞继代核移植,晚期囊胚的发育率急上升(43.84% vs.7.69%),说明继代核移植可促进受体卵对供体核的再程序化。泥鳅和大鳞副泥鳅(Paramisgurnus dabryanus)未受精卵作受体,其移核胚胎发育率无明显区别(7.69% vs.7.02%)。

     

    Abstract: For comparison of hatching rate,fertilized eggs of Misgurnus anguillicaudatus were cultured in different mediums.The results showed that normal and total fries in 1×Holtfreter were the highest,reached 54.8% and 59.2%,respectively.There was no difference in hatching rate of normal fries between 1/2×Holtfreter,1/10×Holtfreter,cold boiled water and changed mediums.They reached 48.0%,49.6%,50.0% and 48.8%,respectively.Rate of abnormal fries was the highest in cold boiled water,reached 8.8%.PBS was not suitable for cultured medium.Mechanic injury had no considerable effects on hatching rate of embryos (73.39% vs.75.70%),but abnormal fries increased considerably (8.8% vs.0).Cultured cells from caudal fin of gynogenetic bighead carp (Aristichthys nobilis) were used as donor cells,and unfertilized eggs of Misgurnus anguillicaudatus were used as recipient eggs.Microsatellite marker analysis showed that the nuclei of nuclear transfer embryos came from the donor nuclei,the nucleus contained in the recipient eggs was rejected or degraded.The passage number of cultured cells critically affected developmental rate of nuclear transfer embryos.With the increase of the passage number,developmental rate of nuclear transfer embryos was decreased.Serial nuclear transfer facilitated reprogramming of nuclear transfer embryos.Developmental rate could be heightened by serial nuclear transfer,especially for the first serial nuclear transfer of somatic cells,rate of late blastula of nuclear transfer embryos increased abruptly from 7.69% to 43.84%.There was no considerable difference in developmental rate of nuclear transfer embryos between Misgurnus anguillicaudatus and Paramisgurnus dabryanus unfertilized recipient eggs (7.69% vs.7.02%).

     

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