钌红对玻璃化冷冻牛卵母细胞线粒体Ca^(2+)的调控研究  

Regulation of Mitochondrial Ca^(2+) in Vitrified Bovine Oocytes by Ruthenium Red

作　　者：王娜[1] 李崇阳[1] 朱化彬[1] 郝海生[1] 王皓宇[1] 闫长亮 杜卫华[1] 王栋[1] 刘岩[1] 庞云渭[1] 赵学明[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]中国牧工商(集团)总公司,北京100070

出　　处：《中国畜牧兽医》2017年第5期1431-1437,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31472100);中国农业科学院"科研英才培育工程";中国农业科学院科技创新工程(ASTIP-IAS06)

摘　　要：玻璃化冷冻会严重损伤哺乳动物卵母细胞的线粒体功能,进而极大地限制了其解冻后的发育能力。为此,本试验设置3个钌红(RR)处理组,即牛卵母细胞用含0.5、1、2μmol/L RR的玻璃化冷冻液进行冷冻,解冻后放入含0.5、1、2μmol/L RR的体外成熟液中继续培养0.5h,同时,新鲜卵母细胞一部分不进行冷冻,一部分用不含RR的冷冻液进行玻璃化冷冻,分别作为新鲜对照组和玻璃化冷冻对照组,然后共检测5组牛卵母细胞线粒体Ca^(2+)水平、ATP含量及孤雌激活后胚胎的发育能力,进而研究RR对玻璃化冷冻牛卵母细胞线粒体Ca^(2+)水平的调控作用。结果显示:(1)玻璃化冷冻显著提高了牛卵母细胞中线粒体Ca^(2+)水平(P<0.05),而2μmol/L RR处理组线粒体Ca^(2+)水平显著低于冷冻对照组(P<0.05),但与新鲜组相比无显著差异(P>0.05);(2)玻璃化冷冻显著降低了牛卵母细胞中ATP含量(P<0.05),2μmol/L RR处理组卵母细胞中ATP含量显著高于冷冻对照组及0.5、1μmol/L RR处理组(P<0.05);(3)玻璃化冷冻对照组卵裂率、囊胚率显著低于新鲜对照组(P<0.05),1μmol/L处理组卵裂率、囊胚率与新鲜对照组相比无显著差异(P>0.05)。综上所述,RR处理能显著抑制解冻后牛卵母细胞线粒体Ca^(2+)流入,保护线粒体功能,提高其发育能力。本试验结果为正向调控玻璃化冷冻卵母细胞线粒体Ca^(2+)水平,进而提高其发育能力,促进玻璃化冷冻卵母细胞的广泛应用提供了参考依据。Studies have shown that the developmental ability of vitrified mammalian oocytes is greatly impacted due to the serious damage of mitochondrial function induced by vitrification process. To solve this problem,in vitro matured bovine oocytes were divided into five groups, three of which were first vitrified using vitrification solution containing 0.5,1, 2 μmol/L rutheni- um red （RR）, then survived oocytes after thawing were cultivated in vitro maturation medium with the same concentration of RR as above for 0.5 h,meanwhile,the remained two groups of oo- cytes were as control, and one group was vitrified using vitrification solution without RR （vitri- fied control group） and the other group had no any treatment （fresh control group）. Then the mi- tochondrial Ca2＋ （mCa2＋） level, ATP content and the developmental capacity of parthenogenetic embryos of oocytes in the five groups were detected to explore the effect of RR on inCa2＋ level of vitrified bovine oocytes. The results showed that：（1）Vitrification significantly increased inCa2＋ concentration in bovine oocytes （P〈0.05）, while RR significantly decreased mCa2＋ concentra tion in vitrified bovine oocytes and there was no significant difference between fresh control group and 2 μmol/L RR group （P〈0.05） ;（2）Vitrification significantly decreased ATP content in bovine oocytes （P〈0.05）,and the ATP content of 2 /amol/L RR group was significantly higher than that of vitrified control group and 0.5,1 μmol/L RR groups （P〈0.05） （3）The cleavage rate and blastocyst rate of fresh control group were significantly higher than those of vitrified control group （P〈0. 05） ,while were no significant difference with 1 htmol/L RR group （P〉0.05）. In conclusion, the results showed that RR treatment could maintain the normal concentration of mCa2＋ in vitrified bovine oocytes,protect the function of mitochondria and improve developmental competence of ooeytes, which would provid theoretical basis to positively r

关 键 词：牛 卵母细胞 玻璃化冷冻 钌红 线粒体Ca2+ 

分 类 号：S823[农业科学—畜牧学]