Department of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
Department of Animal Health and Anatomy, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
Int J Mol Sci. 2020 Oct 13;21(20):7547. doi: 10.3390/ijms21207547.
This study aimed to examine whether the addition of glutathione ethyl ester (GSH-OEt) to the in vitro maturation (IVM) medium would improve the resilience of bovine oocytes to withstand vitrification. The effects of GSH-OEt on spindle morphology, levels of reactive oxygen species (ROS), mitochondrial activity and distribution, and embryo developmental potential were assessed together with the expression of genes with a role in apoptosis (), oxidative-stress pathways (), water channels (), implantation () and gap junctions () in oocytes and their derived blastocysts. Vitrification gave rise to abnormal spindle microtubule configurations and elevated ROS levels. Supplementation of IVM medium with GSH-OEt before vitrification preserved mitochondrial distribution pattern and diminished both cytoplasmic and mitochondrial ROS contents and percentages of embryos developing beyond the 8-cell stage were similar to those recorded in fresh non-vitrified oocytes. Although not significantly different from control vitrified oocytes, vitrified oocytes after GSH-OEt treatment gave rise to similar day 8-blastocyst and hatching rates to fresh non-vitrified oocytes. No effects of GSH-OEt supplementation were noted on the targeted gene expression of oocytes and derived blastocysts, with the exception of , and in derived blastocysts. The addition of GSH-OEt to the IVM medium before vitrification may be beneficial for embryo development presumably as the consequence of additional anti-oxidant protection during IVM.
本研究旨在探讨在体外成熟(IVM)培养基中添加谷胱甘肽乙酯(GSH-OEt)是否能提高牛卵母细胞对玻璃化的抵抗力。研究还评估了 GSH-OEt 对纺锤体形态、活性氧(ROS)水平、线粒体活性和分布以及胚胎发育潜能的影响,同时检测了与凋亡()、氧化应激途径()、水通道()、着床()和间隙连接()相关的基因在卵母细胞及其衍生的囊胚中的表达。玻璃化导致异常的纺锤体微管结构和 ROS 水平升高。在玻璃化前用 GSH-OEt 补充 IVM 培养基可以保持线粒体的分布模式,并降低细胞质和线粒体 ROS 含量,使胚胎发育到 8 细胞阶段以上的比例与新鲜非玻璃化卵母细胞相似。尽管与对照组玻璃化卵母细胞无显著差异,但经 GSH-OEt 处理的玻璃化卵母细胞在第 8 天囊胚和孵化率方面与新鲜非玻璃化卵母细胞相似。除了囊胚中的 、 和 之外,GSH-OEt 补充对卵母细胞和衍生囊胚的靶基因表达没有影响。在玻璃化前将 GSH-OEt 添加到 IVM 培养基中可能对胚胎发育有益,这可能是由于在 IVM 期间额外提供了抗氧化保护。