【作者】 朱玉霞；

【导师】 方南洙；

【作者基本信息】 延边大学 ， 动物遗传育种与繁殖， 2011， 硕士

【摘要】 为了探究延边黄牛卵母细胞Na+,K+-ATP酶的性质,以及卵母细胞体外成熟与Na+,K+-ATP酶活性的相关性,以增加对黄牛卵母细胞体外成熟过程中相关酶活性变化的基础知识,并最终利用相关知识提高延边黄体卵母细胞的体外成熟率。我们首先研究了温度和乌苯苷对延边黄牛卵母细胞Na+,K+-ATP酶活性的影响,进而分别研究了不同等级卵母细胞、不同体外培养时间的A级、E级卵母细胞、体外培养成熟的卵母细胞和体内成熟卵母细胞的Na+,K+-ATP酶活性,对延边黄牛卵母细胞Na+,K+-ATP酶活性的性质进行了初步探讨,并对延边黄牛卵母细胞的体外成熟与其Na+,K+-ATP酶活性的相关性进行了初步研究。试验结果显示,延边黄牛卵母细胞的Na+,K+-ATP酶活性在50℃是达到最大值,在反应温度低于或高于50℃是酶的活性均降低,而且在高于50℃时酶活急剧降低。乌苯苷浓度在2 mmol/L时孤雌卵母细胞完全停止卵裂。A级卵母细胞,裸卵和过成熟卵母细胞三者的Na+,K+-ATP酶活性在统计学上差异不显著。A级,E级和过成熟的卵母细胞中,E级卵母细胞的Na+,K+-ATP酶活性最高。体外培养Oh,6h,12h和24h的A级卵母细胞的Na+,K+-ATP酶活性呈上升趋势,但各组间差异不显著。而体外培养24h的E级卵母细胞的Na+,K+-ATP酶活性显著高于Oh,6h和12h组。体外培养成熟的卵母细胞的Na+,K+-ATP酶活性要显著高于体内成熟及体外培养未成熟的卵母细胞的Na+,K+-ATP酶活性。(结论)由此我们得出结论,延边黄牛卵母细胞的Na+,K+-ATP酶的最适反应温度为50℃,其活力能够被2 mmol/L乌苯苷完全抑制,不同质量的卵母细胞的Na+,K+-ATP酶活性中,裸卵的Na+,K+-ATP酶活性最高。不同等级的卵母细胞的Na+,K+-ATP酶活性不同,体外成熟过程中卵母细胞的Na+,K+-ATP活性呈上升趋势,体外成熟的卵母细胞有较高的Na+,K+-ATP活性。更多还原

【Abstract】 The purpose of this article is to study the character of oocytes in Yanbian Yellow Cattle by researching the effects of temperature on oocytes of Yanbian Yellow Cattle and the max concentration of ouabain, the specific inhibitor of Na+, K+-ATPase, and the effects of granular cell on the active of Na+, K+-ATPase, which are the great foundation of the further study about the relationship between the active of Na+, K+-ATPase and the vitro mature of oocytes of Yanbian Yellow Cattle, and the relativity between the activity of Na+, K+-ATPase and oocytes in vitro mature. The activity of oocytes from different grade was detected, oocytes of Grade A and Grade E were cultured respectively in vitro for Oh,6h,12h,and 24h, and then their Na+, K+-ATPase were determined and compared with each other, the activity of Na+, K+-ATPase of oocytes in vitro mature was compared with that of oocytes in vivo mature. The results showed that the active of Na+, K+-ATPase reached the max value at 50℃, and the active degraded below or over 50℃. The partheno-oocytes cann’t cleavage completely in present of ouabain at 2 mmol/L. The active of Na+, K+-ATPase in grade A group, ova nuda group and hyper-mature group were detected, there were no significant difference among the three groups tested, but the value of ova nuda group was highest. The activity of Na+, K+-ATPase of oocytes of Grade E was the highest in the three groups, Grade A, Grade E and Grade F, however, without significant statistics difference. The activity of Na+, K+-ATPase of Grade A oocytes cultured for Oh,6h,12h and 24h had an increased trend without significant statistics difference. But that of Grade E oocytes cultured for 24 h was significantly higher than 0 h,6 h and 12 h. The activity of Na+, K+-ATPase of oocytes in vitro mature was significantly higher than that of oocytes in vivo mature and immature oocytes in vitro mature.The conclusions were that the optimal react temperature of Na+, K+-ATPase of the oocytes of Yanbian Yellow Cattle was 50℃and the max suppressible concentration of ouabain was 2 mmol/L. The active of Na+, K+-ATPase in ova nuda group was highest. Oocytes in different grade had different activity of Na+, K+-ATPase, and had an increased trend in the process of mature. Oocytes in vitro mature had higher activity of Na+, K+-ATPase.更多还原