Aging-related changes in in vitro-matured bovine oocytes: oxidative stress, mitochondrial activity and ATP content after nuclear maturation.
Aging-related changes in in vitro-matured bovine oocytes: oxidative stress, mitochondrial activity and ATP content after nuclear maturation.
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DOI：
10.1262/jrd.2013-115
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发表时间：
2014-04-24
期刊：
影响因子：
--
通讯作者：
Nagano M
中科院分区：
文献类型：
--
作者：
Koyama K;Kang SS;Huang W;Yanagawa Y;Takahashi Y;Nagano M
本研究的目的是阐明体外成熟牛卵母细胞的衰老相关变化。首先，我们观察了牛卵母细胞在体外成熟（IVM） 22和30-34 h后的受精和胚胎发育情况。体外受精30 ~ 34 h后（开始体外受精40 h左右被精子穿透）的卵母细胞发育成囊胚的率较低（P<0.01），尽管正常受精率与体外受精时间无关。在接下来的实验中，检测IVM作用20、30和40 h后卵母细胞的活性氧（ROS）、线粒体活性和ATP含量。在IVM作用30 h的组中，ROS水平最低。孵育40 h组线粒体活性和ATP含量高于孵育20 h组（P<0.01），孵育30 h组线粒体活性和ATP含量处于中等水平。然后，对体外受精22 h和34 h的卵母细胞胚胎的线粒体活性进行评估。在IVM作用34 h的组中，卵裂球周围经常观察到高极化的线粒体。目前的研究结果表明，长时间体外培养后的卵母细胞线粒体活性高，胚胎发育早期卵裂球外围高极化线粒体的定位可能与老年牛卵母细胞发育能力低有关。
The objective of this research was to clarify the aging-related changes in in vitro-matured bovine oocytes. Firstly, we examined the fertilization and embryonic development of bovine oocytes after 22 and 30–34 h of in vitro maturation (IVM). The oocytes after 30–34 h of IVM (penetrated by sperm at around 40 h after starting IVM) showed a lower developmental rate to blastocysts (P<0.01), although normal fertilization rates were similar regardless of IVM duration. In the next experiment, reactive oxygen species (ROS), mitochondrial activity and ATP content in oocytes after 20, 30 and 40 h of IVM were examined. The lowest level of ROS was found in the group subjected to 30 h of IVM. The mitochondrial activity and ATP content in the group subjected to 40 h of IVM were higher than in the group subjected to 20 h of IVM (P<0.01), and those in the group subjected to 30 h of IVM showed intermediate values. Thereafter, the mitochondrial activities at 3 days after in vitro fertilization in embryos derived from the oocytes subjected to 22 and 34 h of IVM were evaluated. In the group subjected to 34 h of IVM, high-polarized mitochondria were frequently observed at the periphery of blastomeres. The present results suggest that high mitochondrial activity observed in oocytes after prolonged IVM culture and localization of high-polarized mitochondria at the periphery of blastomeres during early embryonic development may be associated with the low developmental competence in aged bovine oocytes.
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