甘氨酸对水貂GV期卵母细胞冷冻保存效率的影响

doi:10.16431/j.cnki.1671-7236.2019.07.014

摘要/Abstract

摘要：

试验旨在探究玻璃化冷冻及培养过程中添加甘氨酸（glycine，Gly）对水貂GV期卵母细胞冷冻解冻后存活率、核发育、线粒体和皮质颗粒分布的影响。试验分为3组：对照组（没有进行冷冻处理）、冷冻组和Gly添加处理组（1 mmol/L Gly）。对玻璃化冷冻解冻后的水貂GV期卵母细胞分别进行平衡恢复3 h和体外成熟培养，采用免疫荧光标记法检测各组GV期卵母细胞线粒体分布的差异及MⅡ期皮质颗粒分布的变化。结果显示，Gly添加处理组卵母细胞在解冻后3 h的存活率与冷冻组相比差异不显著（P>0.05），但显著低于对照组（P<0.05）；Gly添加处理组卵母细胞的减数分裂恢复率显著高于冷冻组（P<0.05），但与对照组相比差异不显著（P>0.05）。免疫荧光结果显示，Gly添加处理组的GV期卵母细胞线粒体正常分布率显著高于冷冻组（P<0.05），但Gly添加处理组和冷冻组的GV期卵母细胞线粒体正常分布率均显著低于对照组（P<0.05）。皮质颗粒分布结果显示，水貂GV期卵母细胞在冷冻后体外成熟培养至MⅡ期时，Gly添加处理组皮质颗粒的正常皮质区分布比例显著高于冷冻组（P<0.05），但Gly添加处理组与冷冻组的正常皮质区分布比例均显著低于对照组（P<0.05）。结果表明，添加Gly可以提高冻融后水貂卵母细胞的减数分裂恢复率，降低冷冻对其线粒体及皮质颗粒的损失。

关键词: 甘氨酸; 水貂; 卵母细胞; 玻璃化冷冻

Abstract:

The purpose of this study was to investigate the effects of glycine (Gly) supplement on the survival rate,nuclear development,mitochondrial distribution and cortical granules distribution in mink oocytes during vitrification and in vitro maturation at germinal vesicle (GV) stage.The experiment was divided into three groups:Control group without vitrification,vitrification group and Gly supplement (1 mmol/L) vitrification group.Mink GV oocytes were cultured for 3 h after vitrification and in vitro maturation,respectively.Immunofluorescence labeling was used to detect the differences of mitochondrial distribution in GV oocytes and the changes of cortical granules distribution in MⅡ oocytes.The results showed that there was no significant difference in the survival rate of oocyte in Gly supplement vitrification group at 3 h after vitrification compared with vitrification group (P>0.05),but it was significantly lower than control group (P<0.05).The meiotic recovery rate of oocytes in Gly supplement vitrification group was significantly higher than vitrification group (P<0.05),but there was no significant difference with control group(P>0.05).The results of immunofluorescence showed that the normal distribution rate of mitochondria in GV oocytes of Gly supplement vitrification group was significantly higher than vitrification group (P<0.05),but the normal mitochondria distribution rate in GV oocytes of Gly supplement vitrification and vitrification groups were significantly lower than control group (P<0.05).The results of cortical granules distribution showed that when mink GV oocytes matured in vitro after vitrification to MⅡ stage,the proportion of cortical granules in Gly supplement vitrification group was significantly higher than vitrification group (P<0.05),but the proportion of cortical granules in Gly supplement vitrification and vitrification grous were significantly lower than control group (P<0.05).The results showed that the Gly supplement in mink oocytes could increase the meiotic recovery rate,and reduce the loss of mitochondria and cortical granules after vitrification.

Key words: glycine; mink; oocyte; vitrification

中图分类号:

S865.2

曹俊国, 李文, 韩玉萍, 李晓霞, 王士勇, 刁云飞, 杨镒峰, 孟庆江, 曹满园, 常彤, 魏海军, 许保增. 甘氨酸对水貂GV期卵母细胞冷冻保存效率的影响[J]. 《中国畜牧兽医》, 2019, 46(7): 1994-2002.

CAO Junguo, LI Wen, HAN Yuping, LI Xiaoxia, WANG Shiyong, DIAO Yunfei, YANG Yifeng, MENG Qingjiang, CAO Manyuan, CHANG Tong, WEI Haijun, XU Baozeng. Effects of Glycine on the Efficiency of Cryopreservation of Mink Oocytes at Germinal Vesicle Stage[J]. , 2019, 46(7): 1994-2002.

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