TMT-based quantitative proteomic analysis of cumulus cells derived from vitrified porcine immature oocytes following in vitro maturation
TMT-based quantitative proteomic analysis of cumulus cells derived from vitrified porcine immature oocytes following in vitro maturation
复制标题
对体外成熟后玻璃化猪未成熟卵母细胞衍生的卵丘细胞进行基于 TMT 的定量蛋白质组学分析。
DOI：
10.1016/j.theriogenology.2020.04.025
复制
发表时间：
2020-08-01
期刊：
影响因子：
2.8
通讯作者：
Wu, Guo-Quan
中科院分区：
文献类型：
--
作者：
Jia, Bao-Yu;Xiang, De-Cai;Wu, Guo-Quan
未成熟卵母细胞在冷冻过程中，其周围的卵丘细胞（CC）不可避免地会受到影响，这可能会对卵母细胞的成熟和发育产生不利影响。到目前为止，对未成熟卵母细胞玻璃化冷冻后CC的分子差异知之甚少。因此，本研究的目的是分析来自玻璃化冷冻猪未成熟卵母细胞体外成熟后的CC的蛋白质谱，使用基于TMT的定量蛋白质组学方法。共鉴定出5910个蛋白质，其中88个蛋白质表达差异显著，上调46个，下调42个。Gene Ontology富集分析显示，细胞周期时相转换、有丝分裂细胞周期时相转换、细胞分化的正调控和卵子发生的调控在生物学过程中均显著下调。经京都基因百科全书和基因组通路分析，部分上调蛋白在TGF-β信号通路和甾体激素相关的4条通路中显著富集。此外，10个选定的蛋白质进行了定量和验证的平行反应监测技术，表明高可靠性的TMT结果。因此，玻璃化冷冻不仅影响了CC的生物学功能，也影响了CC的蛋白质表达，这为了解玻璃化冷冻未成熟卵母细胞成熟质量下降的原因提供了新的视角。(C)2020爱思唯尔公司All rights reserved.
As the immature oocytes are submitted to cryopreservation, their surrounding cumulus cells (CCs) will inevitably suffer, which may have some adverse effects on subsequent oocyte maturation and development. So far, little is known about the molecular differences in CCs of immature oocytes after vitrification. The aim of this study therefore was to analyze the protein profile of CCs derived from vitrified porcine immature oocytes following in vitro maturation, using TMT-based quantitative proteomic approach. A total of 5910 proteins were identified, and 88 of them presented significant difference, with 46 up-regulated and 42 down-regulated proteins. Gene Ontology enrichment analysis revealed that cell cycle phase transition, mitotic cell cycle phase transition, positive regulation of cell differentiation and regulation of oogenesis were significantly down-regulated within the biological process. After Kyoto Encyclopedia of Genes and Genomes pathway analysis, some up-regulated proteins were significantly enriched in TGF-beta signaling pathway and 4 pathways related to steroid hormones. Furthermore, 10 selected proteins were quantified and verified by a parallel reaction monitoring technique, indicating a high reliability of the TMT results. In conclusion, vitrification affects protein profile of CCs as well as their biological functions, which will offer a new perspective to understand the reasons for decline in maturation quality of vitrified immature oocytes. (C) 2020 Elsevier Inc. All rights reserved.