Area of Biochemistry, School of Veterinary Sciences, Institute of Research and Technology in Animal Reproduction, Facultad de Ciencias Veterinarias, University of Buenos Aires, Buenos Aires, Argentina.
Reproduction. 2013 Apr 29;145(5):471-8. doi: 10.1530/REP-13-0017. Print 2013 May.
The knowledge concerning redox and reactive oxygen species (ROS)-mediated regulation of early embryo development is scarce and remains controversial. The aim of this work was to determine ROS production and redox state during early in vitro embryo development in sperm-mediated and parthenogenetic activation of bovine oocytes. Sperm-mediated oocyte activation was carried out in IVF-modified synthetic oviductal fluid (mSOF) with frozen-thawed semen. Parthenogenetic activation was performed in TALP plus ionomycin and then in IVF-mSOF with 6-dimethylaminopurine plus cytochalasin B. Embryos were cultured in IVF-mSOF. ROS and redox state were determined at each 2-h interval (7-24 h from activation) by 2',7'-dichlorodihydrofluorescein diacetate and RedoxSensor Red CC-1 fluorochromes respectively. ROS levels and redox state differed between activated and non-activated oocytes (P<0.05 by ANOVA). In sperm-activated oocytes, an increase was observed between 15 and 19 h (P<0.05). Conversely, in parthenogenetically activated oocytes, we observed a decrease at 9 h (P<0.05). In sperm-activated oocytes, ROS fluctuated throughout the 24 h, presenting peaks around 7, 19, and 24 h (P<0.05), while in parthenogenetic activation, peaks were detected at 7, 11, and 17 h (P<0.05). In the present work, we found clear distinctive metabolic patterns between normal and parthenogenetic zygotes. Oxidative activity and ROS production are an integral part of bovine zygote behavior, and defining a temporal pattern of change may be linked with developmental competence.
关于氧化还原和活性氧（ROS）介导的早期胚胎发育调节的知识很少，而且仍然存在争议。本研究的目的是确定精子介导和孤雌激活牛卵母细胞的早期体外胚胎发育过程中的 ROS 产生和氧化还原状态。精子介导的卵母细胞激活在含有冷冻解冻精液的 IVF 改良合成输卵管液（mSOF）中进行。孤雌激活在 TALP 加离子霉素中进行，然后在含有 6-二甲基氨基嘌呤加细胞松弛素 B 的 IVF-mSOF 中进行。胚胎在 IVF-mSOF 中培养。通过 2'，7'-二氯二氢荧光素二乙酸酯和 RedoxSensor Red CC-1 荧光染料分别在每个 2 小时间隔（激活后 7-24 小时）确定 ROS 和氧化还原状态。ROS 水平和氧化还原状态在激活和未激活的卵母细胞之间存在差异（ANOVA 分析，P<0.05）。在精子激活的卵母细胞中，在 15 至 19 小时之间观察到增加（P<0.05）。相反，在孤雌激活的卵母细胞中，我们在 9 小时观察到减少（P<0.05）。在精子激活的卵母细胞中，ROS 在 24 小时内波动，在 7、19 和 24 小时左右出现峰值（P<0.05），而在孤雌激活中，在 7、11 和 17 小时检测到峰值（P<0.05）。在本研究中，我们发现正常和孤雌胚胎之间存在明显不同的代谢模式。氧化活性和 ROS 产生是牛胚胎行为的一个组成部分，定义变化的时间模式可能与发育能力有关。
