Zhao X-M, Ren J-J, Zhao S-J, Cui L-S, Hao H-S, Wang H-Y, Du W-H, Qin T, Liu Y, Wang D, Zhu H-B
Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Science (IAS), Chinese Academy of Agricultural Sciences (CAAS), Beijing, China.
Reprod Domest Anim. 2014 Aug;49(4):543-549. doi: 10.1111/rda.12305. Epub 2014 Jun 4.
This 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 of the Annexin V assays, as well as measures of viable sperm, early apoptosis, necrotic sperm and mitochondrial membrane potential (∆ψm) showed that the sex-sorted sperm collected from bull A significantly differed from those of the other three bulls (p < 0.05). In addition, the levels of DNA fragmentation in sex-sorted sperm from bull A were significantly lower than those from bulls B and C (p < 0.05). The percentage of cells reaching the cleavage and blastocyst stages in sex-sorted sperm from bull A were significantly greater than those from the other bulls (p < 0.05). A significant positive correlation was observed between viable sperm and the percentage of cells at the cleavage or blastocyst stages (p < 0.05). In contrast, 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.
本研究采用三种染色检测方法(膜联蛋白V、线粒体膜电位(JC-1)和TUNEL),对来自四头不同公牛(A、B、C和D)的性别分选精子进行凋亡的流式细胞术分析。然后评估精子质量与体外受精效率之间的相关性,以确定哪种检测方法能最好地预测体外受精效率。膜联蛋白V检测结果以及活精子、早期凋亡精子、坏死精子和线粒体膜电位(∆ψm)的测量结果表明,从公牛A采集的性别分选精子与其他三头公牛的精子有显著差异(p < 0.05)。此外,公牛A的性别分选精子中的DNA片段化水平显著低于公牛B和C(p < 0.05)。公牛A的性别分选精子中达到卵裂期和囊胚期的细胞百分比显著高于其他公牛(p < 0.05)。观察到活精子与卵裂期或囊胚期细胞百分比之间存在显著正相关(p < 0.05)。相反,早期凋亡精子与卵裂期或囊胚期细胞百分比之间存在负相关(p < 0.05)。总之,这些结果表明,膜联蛋白V检测是预测性别分选牛精子体外受精成功率最可靠的技术。