不同激活方法对猪孤雌胚胎单倍体率的影响

中国农业科学 ›› 2011, Vol. 44 ›› Issue (1): 218-224 .

• 研究简报 • 上一篇

不同激活方法对猪孤雌胚胎单倍体率的影响

张曼玲,赵丽华,周鑫,李荣凤

(内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室)

收稿日期:2010-05-28 修回日期:2010-08-27 出版日期:2011-01-01 发布日期:2011-01-01
通讯作者: 李荣凤

Derivation and Karyotype Analysis of Porcine Haploid Parthenogenetic Embryos

ZHANG Man-ling, ZHAO Li-hua, ZHOU Xin, LI Rong-feng

(内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室)

Received:2010-05-28 Revised:2010-08-27 Online:2011-01-01 Published:2011-01-01
Contact: LI Rong-feng

摘要/Abstract

摘要：

【目的】采用不同的物理或化学方法激活猪卵母细胞,以期获得一种能有效产生猪单倍体孤雌胚胎的激活方法。【方法】试验一分别采用电激活、电激活结合细胞松弛素B(CB)、不同浓度乙醇以及不同作用时间对成熟卵母细胞进行激活处理,试验二检测电激活结合MG-132或Thimerosal和DTT对成熟卵母细胞的激活效果。【结果】试验一显示电激活结合CB组(27.34%)处理后的囊胚率显著高于电激活处理组(16.92%)(P＜0.05),且二者均显著高于所有的乙醇组(P＜0.05),乙醇处理组中以9%乙醇作用11 min效果最佳(10.20%)。试验二显示电激活结合MG-132组的囊胚发育率(24.69%)与电激活结合CB组(27.50%)没有显著差异,二者显著高于电激活结合Thimerosal和DTT组(14.10%)及电激活处理组(17.5%)(P＜0.05)。对所得囊胚进行染色体倍性分析后得出,电激活处理后的囊胚单倍体率(41.7%)与9%乙醇处理组(40%)和电激活结合Thimerosal和DTT处理组(35.3%)无显著差异,但却显著高于电激活结合CB(0)及电激活结合MG-132(6.7%)处理组(P＜0.05)。【结论】综合囊胚发育率和囊胚单倍体比率,电激活、9%乙醇、电激活结合Thimerosal、DTT和电激活结合MG-132处理组获得单倍体囊胚的总效率分别为7.3%(17.5%×41.7%)、4.1%(10.2%×40%)、5.0%(14.1%×35.3%)和1.7%(24.69%×6.7%)。电脉冲处理可能是比较理想的获得猪单倍体孤雌胚胎的激活方法。

关键词: 猪卵母细胞, 体外成熟, 孤雌激活, 单倍体率

Abstract:

【Objective】 In order to obtain high efficient method to produce porcine haploid parthenogenetic embyos, different physical and chemical methods were applied to activate MII porcine oocytes. 【Method】 In experiment 1, electrical activation, a combination of electrical activation and cytochalasin B (CB),and ethanol activation at different concentrations and durations were used to activate porcine oocytes. In experiment 2, electrical activation followed by MG-132 or Thimerosal/DTT treatments were investigated for their effects on porcine oocytes activation. 【Result】 The results of experiment 1 indicated that the development rates of blastocysts in CB group(27.34%)were significantly higher than that of electrical activation alone (16.92%) (P＜0.05), and both of them were significantly higher than that of all ethanol groups (P＜0.05). In all ethanol groups, 9% ethanol treatment for 11 min got the highest blastocyst rate (10.20%) . The results of experiment 2 showed that the rate of blastocysts formation in MG-132 group (24.69%) was not significantly different with that of CB group (27.50%), but they were significantly higher than that of Thimerosal/DTT (14.10%) and electrical activation (17.5%) groups (P＜0.05). The ploidy analysis of blastocysts derived from different groups indicated that the percentages of haploid blastocysts in electrical activation, 9% ethanol and Thimerosal /DTT group were 41.7%, 40%, and 35.3% respectively, they were significantly higher than that of CB (0) and MG-132 (6.7%) groups (P＜0.05)【Conclusion】 Considering both blastocysts formation rate (out of oocytes) and haploid rate (out of total blastocysts), the final haploid blastocyst percentages (out of oocytes) in electrical activation, 9% ethanol, Thimerosal/DTT and MG-132 groups were 7.3% (17.5%×41.7%), 4.1% (10.2%×40%), 5.0% (14.1%×35.3%), and 1.7% (24.69%×6.7%), respectively. The electrical activation might be the best activation way to obtain haploid embryos.

Key words: porcine oocytes, in vitro maturation, parthenogenetic activation, rate of haploid

张曼玲,赵丽华,周鑫,李荣凤 . 不同激活方法对猪孤雌胚胎单倍体率的影响
[J]. 中国农业科学, 2011, 44(1): 218-224 .

ZHANG Man-ling,ZHAO Li-hua,ZHOU Xin,LI Rong-feng
.

Derivation and Karyotype Analysis of Porcine Haploid Parthenogenetic Embryos

[J]. Scientia Agricultura Sinica, 2011, 44(1): 218-224 .

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不同激活方法对猪孤雌胚胎单倍体率的影响

Derivation and Karyotype Analysis of Porcine Haploid Parthenogenetic Embryos

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