Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Sci Rep. 2018 Jun 21;8(1):9477. doi: 10.1038/s41598-018-27829-9.
Cumulus cells are essential for nutrition of oocytes during maturation. In the absence of cumulus cells during maturation, oocyte developmental competence is severely compromised. In this study, we matured bovine cumulus-oocyte-complexes (COCs) for 8 h, the cumulus cells were removed and denuded oocytes were further matured for 15 h in either the medium conditioned by the initial 8 h culture, or in fresh unconditioned medium. Denuded oocytes that completed maturation in COC-conditioned medium demonstrated better developmental potential than denuded oocytes that completed maturation in standard maturation medium. An inventory was made of the metabolites secreted by COCs into the maturation medium during the first 8 h, from 8 to 23 h, and during an entire 23 h maturation protocol; the metabolomic changes in the cumulus cells during maturation were also investigated. In maturation medium, 173 biochemical components were detected compared to 369 different metabolites in cumulus cells. Significant changes in metabolomic components were evident in maturation medium and in cumulus cells during maturation, with most of the changes related to amino acid, carbohydrate, and lipid metabolism. The importance of two detected biochemicals, creatine and carnitine, for oocyte maturation was further investigated. The presence of carnitine, but not creatine during oocyte in vitro maturation improved the developmental competence of denuded oocytes.
卵丘细胞在卵母细胞成熟过程中对于营养物质的供给至关重要。在卵母细胞成熟过程中去除卵丘细胞,会严重损害卵母细胞的发育能力。在这项研究中,我们对牛卵丘-卵母细胞复合物(COC)进行了 8 小时的成熟培养,然后去除卵丘细胞,让裸露的卵母细胞在初始 8 小时培养的条件培养基或新鲜未处理的培养基中进一步成熟 15 小时。与在标准成熟培养基中完成成熟的裸露卵母细胞相比,在 COC 条件培养基中完成成熟的裸露卵母细胞显示出更好的发育潜能。我们对 COC 在最初的 8 小时、8 至 23 小时以及整个 23 小时成熟过程中分泌到成熟培养基中的代谢产物进行了清单制作;还研究了卵丘细胞在成熟过程中的代谢变化。与卵丘细胞中检测到的 369 种不同代谢物相比,在成熟培养基中检测到了 173 种生化成分。在成熟培养基中和卵丘细胞在成熟过程中的代谢物成分都发生了显著变化,大多数变化与氨基酸、碳水化合物和脂质代谢有关。进一步研究了两种检测到的生化物质肌酸和肉碱对卵母细胞成熟的重要性。在体外成熟过程中,肉碱的存在而不是肌酸的存在提高了裸露卵母细胞的发育能力。