Roy Pantu Kumar, Qamar Ahmad Yar, Fang Xun, Hassan Bahia Mohamed Salih, Cho Jongki
College of Veterinary Medicine, Chungnam National University, Daejeon, 34134, Republic of Korea.
College of Veterinary Medicine, Chungnam National University, Daejeon, 34134, Republic of Korea; College of Veterinary and Animal Sciences, Jhang, 35200, Pakistan.
Theriogenology. 2020 Sep 15;154:24-30. doi: 10.1016/j.theriogenology.2020.05.011. Epub 2020 May 15.
This study was performed to explore the effects of cobalamin treatment during in vitro maturation (IVM) of porcine oocytes. Specifically, the effects of cobalamin exposure on nuclear and cytoplasmic maturation of oocytes, diameter, glutathione (GSH) and reactive oxygen species (ROS) levels of matured oocytes, as well as development and gene expression of porcine parthenogenetic and cloned embryos were assessed. Cumulus-oocyte complexes were exposed to 200 pM cobalamin for 22 h or incubated for 22 h without cobalamin (controls). The mean diameter of cobalamin-treated oocytes was greater than that of control oocytes (160.0 vs. 154.5 μm; p < 0.05). GSH level increased but ROS level decreased in the cobalamin-treated oocyte group. Parthenogenetic embryos derived from cobalamin-treated oocytes showed improved oocyte maturation (91.3% vs. 83.8%), cleavage (88.9% vs. 82.1%), and blastocyst formation (38.7% vs. 31.9%) rates compared with control embryos (p < 0.05). Similarly, cloned embryos derived from cobalamin-treated oocytes showed higher oocyte maturation (89.2% vs. 82.2%), cleavage (87.5% vs. 80.3%), and blastocyst formation (30.0% vs. 23.8%) rates than control embryos (p < 0.05). Furthermore, in parthenogenetic and cloned embryos, total cell number, inner cell mass (ICM), trophectoderm (TE) expression, and ICM: TE ratio were higher in the cobalamin-treated group compared to that in the control group (p < 0.05). Cloned embryos in the cobalamin-treated group showed higher mRNA expression levels of POU5F1, DPPA2, and NDP52IL than control group embryos. Together, these results demonstrate that cobalamin treatment during IVM improves the developmental competence of porcine oocytes by neutralizing the free radicals produced in the IVM medium.
本研究旨在探讨钴胺素处理对猪卵母细胞体外成熟(IVM)的影响。具体而言,评估了钴胺素暴露对卵母细胞核成熟和细胞质成熟、成熟卵母细胞的直径、谷胱甘肽(GSH)和活性氧(ROS)水平,以及猪孤雌胚胎和克隆胚胎的发育及基因表达的影响。将卵丘 - 卵母细胞复合体暴露于200 pM钴胺素中22小时,或在无钴胺素的情况下孵育22小时(对照组)。经钴胺素处理的卵母细胞的平均直径大于对照卵母细胞(160.0对154.5μm;p <0.05)。在经钴胺素处理的卵母细胞组中,GSH水平升高而ROS水平降低。与对照胚胎相比,源自经钴胺素处理的卵母细胞的孤雌胚胎显示出更好的卵母细胞成熟率(91.3%对83.8%)、卵裂率(88.9%对82.1%)和囊胚形成率(38.7%对31.9%)(p <0.05)。同样,源自经钴胺素处理的卵母细胞的克隆胚胎显示出比对照胚胎更高的卵母细胞成熟率(89.2%对82.2%)、卵裂率(87.5%对80.3%)和囊胚形成率(30.0%对23.8%)(p <…