College of Veterinary Medicine, Yunnan Agricultural University, Kunming, Yunnan, 650201, China.
Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Yunnan Animal Science and Veterinary Institute, Kunming, Yunnan, 650224, China.
Theriogenology. 2020 Aug;152:8-17. doi: 10.1016/j.theriogenology.2020.04.025. Epub 2020 Apr 23.
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.
由于不成熟的卵母细胞需要进行冷冻保存，其周围的卵丘细胞（CCs）不可避免地会受到影响，这可能对随后的卵母细胞成熟和发育产生一些不利影响。到目前为止，对于玻璃化处理后的不成熟卵母细胞的 CCs 分子差异知之甚少。因此，本研究旨在使用基于 TMT 的定量蛋白质组学方法分析玻璃化处理后的猪不成熟卵母细胞体外成熟后 CCs 的蛋白质谱。共鉴定出 5910 种蛋白质，其中 88 种蛋白质呈现显著差异，其中 46 种上调，42 种下调。GO 富集分析显示，生物过程中细胞周期相位转变、有丝分裂细胞周期相位转变、细胞分化的正调控和卵发生的调控显著下调。KEGG 通路分析后，一些上调的蛋白质在 TGF-β信号通路和 4 个与甾体激素相关的通路中显著富集。此外，通过平行反应监测技术对 10 种选定的蛋白质进行了定量验证，表明 TMT 结果具有很高的可靠性。总之，玻璃化处理会影响 CCs 的蛋白质谱及其生物学功能，这为理解玻璃化处理不成熟卵母细胞成熟质量下降的原因提供了新的视角。
