性控分离精子凋亡程度对牛体外受精的影响  

Effects of the Degree of Apoptosis in Sexed Sperms on Bovine Fertilization in vitro

作　　者：郝海生[1] 任晶晶[1] 赵学明[1] 王玲玲 杜卫华[1] 王长法 朱化彬[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]山东奥克斯生物技术有限公司,济南250000

出　　处：《畜牧兽医学报》2014年第6期893-900,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：现代农业产业技术体系建设专项资金(CARS-37);十二五科技支撑项目(2011BAD19B04-6);引智项目(Y20110326005)

摘　　要：采用Annexin-V、JC-1和TUNEL对A、B、C、D 4头种公牛性控X冻精进行了凋亡分析,并对凋亡分析结果与性控精液体外受精卵裂率和囊胚率的关系进行了相关分析。结果表明:(1)公牛A的活精子(AN-·PI-)百分率(89.30%±3.04%)高于其他公牛((74.39%±3.24%)^(81.17%±4.25%)),差异显著(P<0.05),而其DNA片段化精子百分率(12.24%±1.25%)低于其他公牛((19.09%±1.27%)^(27.56%±3.17%)),差异显著(P<0.05);(2)公牛A的X冻精体外受精(IVF)卵裂率、囊胚率((76.82%±3.00%)^(20.45%±1.44%))均高于其他3头公牛((22.00%±6.85%)^(31.88%±6.69%)),(3.33%±2.67%)^(4.00%±3.26%)),差异显著(P<0.05);(3)活精子百分率与其IVF卵裂率和囊胚率呈正相关,而DNA片段化百分率与之呈负相关。综上表明:Annexin-V分析能够准确地预测牛X性控冻精的体外受精效率,这对于促进牛性控体外胚胎的生产效率有着重要意义。The present study utilized three staining assays(Annexin-V,mitochondrial membrane potential(JC-1),and TUNEL)for flow cytometric analysis of apoptosis in sex-sorted sperm from four different bulls(A,B,C and D).Correlations between sperm quality and IVF efficiency were then assessed to determine which assay provided the best prediction of IVF efficiency.The results showed that:(1)the percentage of viable sperm(AN-·PI-)in X-sorted semen from bull A(89.30%±3.04%)was significantly higher than that from other bulls((74.39%±3.24%)-(81.17%±4.25%);P<0.05),while the percentage of DNA fragmentation sperm from bull A(12.24%±1.25%)was significantly lower than that from other bulls((19.09%±1.27%)-(27.56%±3.17%);P<0.05);(2)the percentage of cells reaching the cleavage and blastocyst stages after in vitro fertilization(IVF)in sex-sorted sperm from bull A((76.82%±3.00%),(20.45%±1.44%))were significantly greater than those from the other bulls((22.00% ±6.85%)-(31.88%±6.69%),(3.33%±2.67%)-(4.00%±3.26%);P<0.05);(3)a significant positive correlation was observed between viable sperm and the percentage of cells at the cleavage or blastocyst stages(P<0.05)and a negative correlation was found between early apoptotic sperm and the percentage of cells at the cleavage or blastocyst stages(P<0.05).In conclusion,these results indicated that the Annexin-V assay was the most reliable technique for the prediction of the IVF success of sex-sorted bovine sperm.

关 键 词：体外受精 不同种公牛 ANNEXIN-V JC-1 TUNEL 精子凋亡 精液质量 

分 类 号：S823[农业科学—畜牧学]