猪卵母细胞孤雌激活方法及影响因素的研究  

Studies on Methods and Influencing Factors of Porcine Oocyte Activation

作　　者：孙兴参[1] 岳奎忠[1] 罗明久[2] 谭景和[2] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]山东农业大学动物科技学院,泰安271018

出　　处：《中国农业科学》2004年第3期431-435,共5页Scientia Agricultura Sinica

基　　金：国家"973"资助项目(G200016107)

摘　　要：研究了离子霉素、电刺激、二硫苏糖醇、6-DMAP和硫柳汞等单独或结合使用及极体形态对猪卵母细胞的激活率、原核类型及激活卵卵裂的影响。结果表明:(1) 单独用离子霉素(88.9%)或电脉冲(80.0%)处理卵母细胞的激活率与二者同6-DMAP结合处理的激活率(84.3%和79.1%)差异不显著,但6-DMAP处理明显(P<0.05)提高电激活卵的1PN比率,而对离子霉素激活卵作用不明显。(2) DTT后处理显著提高硫柳汞处理对卵母细胞的激活率(由4.6%提高到82.6%)。(3) 极体皱缩(85.4%)和极体碎裂(86.2%)卵母细胞的电激活率显著高于极体完整卵母细胞(58.9%)。极体完整卵母细胞形成1PN的比率(78.8%)显著高于,而形成2PN的比率(18.2%)显著低于极体皱缩和碎裂卵母细胞。(4) 电脉冲结合6-DMAP激活卵的卵裂率(52.4%)明显高于离子霉素结合6-DMAP激活卵(23.0%),但与单纯电脉冲刺激激活卵的卵裂率(46.1%)没有明显差异。The effects of ionomycin, electrical pulses, dithiothreitol (DTT), 6-DMAP and thimerosal, when used eitheralone or in combinations, on the activation rate, pronuclear type and cleavage of porcine oocyte were studied. The resultsare as follows: (1) The activation rates of oocytes treated with ionomycin (88.9%) or pulsed (80.0%) alone were not differentsignificantly from each other and from those in oocytes treated in combination with 6-DMAP (84.3% and 79.1%, respectively).While 6-DMAP improved electrical activation (increased the proportion of oocytes with 1 pronucleus), it had no effect onoocyte activation by ionomycin.(2)Post-treatment with DTT significantly enhanced the thimerosal activation of porcineoocytes with activation rates increased from 4.6% to 82.6%.(3)When pulsed, the activation rates of oocytes withcrimped (85.4%) or fragmented first polar bodies (86.2%) were significantly higher than those of oocytes with intact polarbodies (58.9%); the ratio of 1 PN activated oocytes (78.8%) was significantly higher, but that of 2PN oocytes (18.2%) wassignificantly lower in oocytes with intact polar bodies.(4) The cleavage rate of oocytes activated by electrical stimulus(52.4%) was significantly higher than that in oocytes activated by ionomycin (23.0%) when combined with 6-DMAP, but itwas not significantly different from that in oocytes activated by electrical stimulus alone (46.1%).

关 键 词：猪 卵母细胞 孤雌激活 离子霉素 电刺激 二硫苏糖醇 硫柳汞 6—DMAP 极体形态 核移植 

分 类 号：S828[农业科学—畜牧学]