Lee Joohyeong, Kim Eunhye, Hwang Seon-Ung, Cai Lian, Kim Mirae, Choi Hyerin, Oh Dongjin, Lee Eunsong, Hyun Sang-Hwan
Laboratory of Veterinary Embryology and Bio-technology (VETEMBIO), Veterinary Medical Center and College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Korea.
Institute of Stem Cell & Regenerative Medicine (ISCRM), Chungbuk National University, Cheongju 28644, Korea.
Animals (Basel). 2021 Apr 6;11(4):1034. doi: 10.3390/ani11041034.
This study aimed to examine the effects of treatment with glucuronic acid (GA) and N-acetyl-D-glucosamine (AG), which are components of hyaluronic acid (HA), during porcine oocyte in vitro maturation (IVM). We measured the diameter of the oocyte, the thickness of the perivitelline space (PVS), the reactive oxygen species (ROS) level, and the expression of cumulus cell expansion and ROS-related genes and examined the cortical granule (CG) reaction of oocytes. The addition of 0.05 mM GA and 0.05 mM AG during the first 22 h of oocyte IVM significantly increased oocyte diameter and PVS size compared with the control (non-treatment). The addition of GA and AG reduced the intra-oocyte ROS content and improved the CG of the oocyte. GA and AG treatment increased the expression of and in cumulus cells and and in oocytes. In both the chemically defined and the complex medium (Medium-199 + porcine follicular fluid), oocytes derived from the GA and AG treatments presented significantly higher blastocyst rates than the control after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). In conclusion, the addition of GA and AG during IVM in pig oocytes has beneficial effects on oocyte IVM and early embryonic development after PA and SCNT.
本研究旨在探讨透明质酸(HA)的组成成分葡萄糖醛酸(GA)和N-乙酰-D-葡萄糖胺(AG)在猪卵母细胞体外成熟(IVM)过程中的处理效果。我们测量了卵母细胞的直径、卵周隙(PVS)的厚度、活性氧(ROS)水平以及卵丘细胞扩展和ROS相关基因的表达,并检测了卵母细胞的皮质颗粒(CG)反应。与对照组(未处理)相比,在卵母细胞IVM的前22小时添加0.05 mM GA和0.05 mM AG显著增加了卵母细胞直径和PVS大小。GA和AG的添加降低了卵母细胞内ROS含量并改善了卵母细胞的CG。GA和AG处理增加了卵丘细胞中 和 的表达以及卵母细胞中 和 的表达。在化学成分明确的培养基和复合培养基(M199 + 猪卵泡液)中,经GA和AG处理的卵母细胞在孤雌激活(PA)和体细胞核移植(SCNT)后,其囊胚率均显著高于对照组。总之,在猪卵母细胞IVM过程中添加GA和AG对卵母细胞IVM以及PA和SCNT后的早期胚胎发育具有有益影响。