猪卵母细胞在GV期与MⅡ期的冷冻保存效果比较研究

发布时间：2018-07-13 10:13

【摘要】：为比较猪卵母细胞在GV期与MⅡ期的冷冻保存效果,试验在这两个成熟阶段对其进行玻璃化冷冻,GV期卵母细胞解冻后培养至成熟,MⅡ期卵母细胞解冻后恢复2h,然后采用免疫荧光标记、Western blotting和链霉蛋白酶溶解方法分别检测它们的皮质颗粒分布、CD9蛋白表达水平和透明带消化时间上的差异。结果表明,GV期卵母细胞在解冻后2h的存活率显著低于MⅡ期卵母细胞(P0.05),但极体排出率与对照卵母细胞无明显差异(P0.05);在冷冻MⅡ期卵母细胞中,皮质颗粒的皮质区分布比例和CD9的蛋白表达水平显著下降(P0.05),但冷冻GV期卵母细胞经体外成熟后则无明显变化(P0.05);冷冻GV期与MⅡ期卵母细胞均不会影响透明带的消化时间(P0.05)。由此可见,猪卵母细胞在GV期的冷冻存活率虽然较MⅡ期低,但其体外成熟后极体排出率、皮质颗粒分布和CD9蛋白表达水平均未受到冷冻的影响。
[Abstract]:In order to compare the cryopreservation effect of porcine oocytes during the period of GV and M II, the test was vitrified at these two mature stages, GV oocytes were cultured to mature after thawing, and 2H was restored after thawing of M II oocytes. Then, immunofluorescence markers, Western blotting and streptomycin dissolution methods were used to detect them respectively. The distribution of cortical granules, the expression level of CD9 protein and the difference in the digestion time of the zona pellucida showed that the survival rate of 2h in GV oocytes was significantly lower than that of M II oocytes (P0.05) after thawing, but the rate of polar body discharge was not significantly different from that of the control oocytes (P0.05). The cortical distribution ratio of the cortical granules in the frozen M II oocytes was compared with that of the control oocytes. The expression level of protein and CD9 decreased significantly (P0.05), but there was no significant change in the frozen GV oocyte after maturation in vitro (P0.05). The frozen GV stage and M II oocyte did not affect the digestion time of the zona pellucida (P0.05). Thus, the survival rate of porcine oocytes in GV phase was lower than that of M stage II, but it was mature in vitro. Body excretion rate, cortical granule distribution and CD9 protein expression were not affected by freezing.
【作者单位】：云南省畜牧兽医科学院云南省畜禽遗传资源保护和种质创新工程实验室;
【基金】：国家自然科学基金(31402081) 云南省应用基础研究计划项目(2014FB162) 云南省高新技术产业发展专项项目(201404)
【分类号】：S828

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