玻璃化冻存对驴卵母细胞超微结构的影响

doi:10.16431/j.cnki.1671-7236.2018.01.019

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (1): 147-153.doi: 10.16431/j.cnki.1671-7236.2018.01.019

玻璃化冻存对驴卵母细胞超微结构的影响

庞玉娟^1,2, 杜兴雨^1,2, 李强^1,2, 张志鹏^1,2,3, 张焱如^1,2, 潘庆杰³, 曹俊伟^1,2

1. 内蒙古农业大学生命科学学院, 呼和浩特 010018;
2. 内蒙古自治区生物制造重点实验室, 呼和浩特 010018;
3. 青岛农业大学动物科技学院, 青岛 266109

收稿日期:2017-07-17 出版日期:2018-01-20 发布日期:2018-01-20
通讯作者: 曹俊伟 E-mail:caojunwei1970@163.com
作者简介:庞玉娟(1992-),女,内蒙古乌兰察布人,硕士生,研究方向:动物繁殖育种与胚胎工程,E-mail:1904721636@qq.com
基金资助:
国家自然科学基金（31460292）

Effects of Vitrification Freezing on Oocytes Ultrastructure in Equus asinus

PANG Yujuan^1,2, DU Xingyu^1,2, LI Qiang^1,2, ZHANG Zhipeng^1,2,3, ZHANG Yanru^1,2, PAN Qingjie³, CAO Junwei^1,2

1. College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Key Laboratory of Inner Mongolia Autonomous Region Biological Manufacturing, Hohhot 010018, China;
3. College of Animal Sciences and Technology, Qingdao Agricultural University, Qingdao 266109, China

Received:2017-07-17 Online:2018-01-20 Published:2018-01-20

摘要/Abstract

摘要：

试验旨在探究玻璃化冷冻对驴卵母细胞发育的影响，寻求驴卵母细胞冷冻的最佳条件。通过对不同发育时期的驴卵母细胞进行玻璃化冷冻，冷冻复苏后分别进行成熟培养和孤雌激活，并对GV期未冷冻组（对照组）、GV期冷冻组、IVM-M Ⅱ冷冻组卵母细胞微丝和线粒体超微结构进行免疫荧光标记，统计冷冻复苏后卵母细胞形态正常率、成熟率、孤雌激活卵裂率、超微结构正常率。结果表明，GV期冷冻组卵母细胞的形态正常率与GV期未冷冻组（对照组）间无显著差异（P>0.05），成熟率和卵裂率均显著低于对照组（P<0.05）；IVM-M Ⅱ冷冻组的卵裂率显著低于对照组（P<0.05），且卵裂后细胞发育受到阻滞。冷冻组微丝在皮质区分布明显减少的卵母细胞数目增多，冷冻组卵母细胞的线粒体数量明显低于对照组，由此可以说明冷冻对卵母细胞超微结构有损伤，从而导致复苏后成熟率下降，影响卵母细胞的受精和体外发育，且GV期冷冻组较IVM-M Ⅱ冷冻组在微丝与线粒体结构上有较小损伤，发育状态较好。

关键词: 驴; 卵母细胞; 玻璃化冷冻; 微丝; 线粒体

Abstract:

This study was aimed to evaluate the effect of vitrification freezing on the development of oocytes in Equus asinus, and seek the best freezing conditions of oocytes in Equus asinus. The oocytes of Equus asinus in different development stages were vitrified and frozen, after the resuscitation, the mature culture and parthenogenetic activation were performed, respectively. The oocytes were stained with microfilaments and mitochondrial ultrastructure, and the oocytes were divided into GV stage without freezing group (control group), GV stage freezing group, IVM-M Ⅱ freezing group. The normal rate of oocyte morphology, maturation rate, the cleavage rate of partial activation, the normal rate of ultrastructural were counted. The results showed that there was no significant difference in the normal rate of morphology between GV stage freezing group and GV stage without freezing (P>0.05), and the maturation rate and cleavage rate were significantly lower than those in control group (P<0.05), the cleavage rate of IVM-M Ⅱ freezing group was significantly lower than that of control group (P<0.05), and the cell development was blocked after cleavage. The distribution of microfilaments in the cortex of most oocytes in freezing group was significantly reduced, and the number of mitochondria in freezing group was significantly lower than that in control group, so it could be explained that freezing caused damage to the oocyte's ultrastructure, resulting in decreased resuscitation after the maturity rate, affecting the oocyte fertilization and in vitro development, and the GV stage freezing group had less damage to microfilaments and mitochondrial structure than IVM-M Ⅱ freezing group, and the developmental status was better.

Key words: Equus asinus; oocyte; vitrification freezing; microfilaments; mitochondria

中图分类号:

S822.3

庞玉娟, 杜兴雨, 李强, 张志鹏, 张焱如, 潘庆杰, 曹俊伟. 玻璃化冻存对驴卵母细胞超微结构的影响[J]. 《中国畜牧兽医》, 2018, 45(1): 147-153.

PANG Yujuan, DU Xingyu, LI Qiang, ZHANG Zhipeng, ZHANG Yanru, PAN Qingjie, CAO Junwei. Effects of Vitrification Freezing on Oocytes Ultrastructure in Equus asinus[J]. , 2018, 45(1): 147-153.

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玻璃化冻存对驴卵母细胞超微结构的影响

Effects of Vitrification Freezing on Oocytes Ultrastructure in Equus asinus

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