Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University (Chengdu Campus), Wenjiang, PR China.
Mol Reprod Dev. 2013 Jun;80(6):451-9. doi: 10.1002/mrd.22181. Epub 2013 May 20.
This study was conducted to investigate the effect of vitrification of bovine metaphase-II (MII) oocytes on CD9 expression and fertilization capacity. Surviving vitrified/warmed oocytes were used to detect CD9 distribution (fluorescence microscopy), CD9 mRNA (qRT-PCR), and CD9 protein expression (Western blot), and to analyze in vitro fertilization rates (number of sperm bound to or that penetrated the oocytes) after removing the zona pellucida. Fresh oocytes acted as control. The experimental results showed that the vitrification/warming procedures significantly decreased CD9 expression at the mRNA and protein levels, and changed the CD9 distribution pattern in bovine oocytes. After fertilization in vitro, the average number of sperm binding and penetration of vitrified oocytes were significantly lower than those of the non-vitrified oocytes. In conclusion, vitrification of bovine oocytes caused a decrease in CD9 expression at the mRNA and protein levels, and an alteration of CD9 distribution pattern, which may have resulted in lowered fertilization capacity.
本研究旨在探讨玻璃化冷冻对牛中期 II (MII) 卵母细胞 CD9 表达和受精能力的影响。使用存活的玻璃化/解冻卵母细胞检测 CD9 分布（荧光显微镜）、CD9 mRNA（qRT-PCR）和 CD9 蛋白表达（Western blot），并在去除透明带后分析体外受精率（与卵母细胞结合或穿透的精子数量）。新鲜卵母细胞作为对照。实验结果表明，玻璃化/解冻程序显著降低了牛卵母细胞中 CD9 的 mRNA 和蛋白水平的表达，并改变了 CD9 的分布模式。体外受精后，玻璃化卵母细胞的平均精子结合和穿透数量明显低于非玻璃化卵母细胞。总之，牛卵母细胞的玻璃化处理导致 CD9 的 mRNA 和蛋白水平表达降低，CD9 分布模式改变，可能导致受精能力降低。
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