Kim Keun Jung, Chun Ju Lan, Lee Kyung-Bon, Lee Ji Hye, Park Kang-Sun, Han Kil Woo, Lee Bo Myeong, Kim Eun Young, Kim Jin Man, Kim Min Kyu
Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, 305-764, Republic of Korea.
Department of Biology Education, College of Education, Chonnam National University, Gwangju, 500-757, Republic of Korea.
J Assist Reprod Genet. 2016 Jul;33(7):939-48. doi: 10.1007/s10815-016-0729-x. Epub 2016 May 17.
The aim of this study is to investigate the effect of acteoside, an antioxidant, on in vitro maturation (IVM) of oocytes to improve early parthenogenetic embryonic developmental competence.
Porcine immature oocytes (total 770) were cultured in IVM medium with acteoside at various concentrations, 0 (control), 10, 30, and 50 μM. Each group was assessed for maturation and subsequent development rates, reactive oxygen species (ROS) level (15 oocytes per group and four independent experiments performed), ultrastructure observation (15 oocytes per group), mitochondrial activity (30 oocytes per groups and three independent experiments performed), and expression patterns of apoptosis-related genes (100 expended parthenogenetic embryos per group and three independent experiment performed). Main outcome measures were the rates of IVM, blastocyst formation, ROS, mitochondria, and expression of apoptosis-related genes in oocytes treated with acteoside.
RESULT(S): Addition of acteoside during IVM did not change the maturation efficiency of oocytes but improved the rate of blastocyst formation with significantly decreased ROS level. Moreover, in acteoside-treated oocytes, cytoplasmic maturation was improved with morphologically uniform distribution of mitochondria and lipid droplets in cytoplasm. Acteoside supplementation also increased the mRNA expression levels of antiapoptotic genes and reduced those of pro-apoptotic genes.
CONCLUSION(S): Acteoside supplementation in IVM medium improves the oocyte quality and subsequent development of pre-implantation embryos that would eventually contribute to produce embryos with high embryonic development competence.
本研究旨在探讨抗氧化剂毛蕊花糖苷对卵母细胞体外成熟(IVM)的影响,以提高早期孤雌胚胎发育能力。
将猪未成熟卵母细胞(共770个)培养于含有不同浓度毛蕊花糖苷(0(对照)、10、30和50 μM)的IVM培养基中。对每组进行成熟率和后续发育率评估、活性氧(ROS)水平检测(每组15个卵母细胞,进行4次独立实验)、超微结构观察(每组15个卵母细胞)、线粒体活性检测(每组30个卵母细胞,进行3次独立实验)以及凋亡相关基因表达模式分析(每组100个孤雌胚胎,进行3次独立实验)。主要观察指标为经毛蕊花糖苷处理的卵母细胞的IVM率、囊胚形成率、ROS水平、线粒体情况及凋亡相关基因的表达。
在IVM过程中添加毛蕊花糖苷并未改变卵母细胞的成熟效率,但提高了囊胚形成率,且ROS水平显著降低。此外,在经毛蕊花糖苷处理的卵母细胞中,细胞质成熟得到改善,线粒体和脂滴在细胞质中形态均匀分布。补充毛蕊花糖苷还增加了抗凋亡基因的mRNA表达水平,降低了促凋亡基因的表达水平。
在IVM培养基中补充毛蕊花糖苷可提高卵母细胞质量和植入前胚胎的后续发育能力,最终有助于产生具有高胚胎发育能力的胚胎。