Jia Baoyu, Xiang Decai, Fu Xiangwei, Shao Qingyong, Hong Qionghua, Quan Guobo, Wu Guoquan
College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.
Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Yunnan Animal Science and Veterinary Institute, Kunming, China.
Front Cell Dev Biol. 2020 Dec 23;8:614577. doi: 10.3389/fcell.2020.614577. eCollection 2020.
Cryopreservation of immature germinal vesicle (GV) oocytes is a promising strategy in pigs but still results in reduced oocyte quality due to inevitable cryodamages. Recently, there has been more focus on the molecular changes of oocytes after vitrification, but the alteration in the proteome level remains elusive. The aim of this study therefore was to decipher the proteomic characteristics of porcine GV oocytes following vitrification and maturation (IVM) by using tandem mass tag (TMT)-based quantitative approach and bioinformatics analysis. A total of 4,499 proteins were identified, out of which 153 presented significant difference. There were 94 up-regulated and 59 down-regulated proteins expressed differentially in the vitrified oocytes. Functional classification and enrichment analyses revealed that many of these proteins were involved in metabolism, signal transduction, response to stimulus, immune response, complement, coagulation cascades, and so on. Moreover, a parallel reaction monitoring technique validated the reliability of TMT data through quantitative analysis for 10 candidate proteins. In conclusion, our results provided a novel perspective of proteomics to comprehend the quality change in the vitrified porcine GV oocytes after IVM.
未成熟生发泡（GV）期猪卵母细胞的冷冻保存是一种很有前景的策略，但由于不可避免的冷冻损伤，其卵母细胞质量仍会下降。近年来，人们更多地关注了玻璃化冷冻后卵母细胞的分子变化，但蛋白质组水平的改变仍不清楚。因此，本研究的目的是通过基于串联质谱标签（TMT）的定量方法和生物信息学分析，解析玻璃化冷冻及体外成熟（IVM）后猪GV期卵母细胞的蛋白质组学特征。共鉴定出4499种蛋白质，其中153种存在显著差异。在玻璃化冷冻的卵母细胞中，有94种蛋白质上调，59种蛋白质下调。功能分类和富集分析表明，这些蛋白质中有许多参与代谢、信号转导、应激反应、免疫反应、补体、凝血级联反应等。此外，平行反应监测技术通过对10种候选蛋白质的定量分析验证了TMT数据的可靠性。总之，我们的结果为理解玻璃化冷冻的猪GV期卵母细胞IVM后的质量变化提供了蛋白质组学的新视角。
