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基于iTRAQ的超排刺激后羔羊和成年母羊颗粒细胞蛋白质组学分析。

iTRAQ-based proteomic profiling of granulosa cells from lamb and ewe after superstimulation.

作者信息

Lin Jiapeng, Wu Yangsheng, Han Bing, Chen Ying, Wang Liqin, Li Xiaolin, Liu Mingjun, Huang Juncheng

机构信息

College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China; Biotechnology Research Institute, Xinjiang Academy of Animal Science, Urumqi, 830000, China.

Biotechnology Research Institute, Xinjiang Academy of Animal Science, Urumqi, 830000, China.

出版信息

Theriogenology. 2017 Oct 1;101:99-108. doi: 10.1016/j.theriogenology.2017.06.014. Epub 2017 Jun 10.

Abstract

The number of oocytes obtained from lambs after FSH treatment is far greater than those acquired from adult ewes. However, these oocytes typically have reduced viability in comparison with adult ewe oocytes. However, the molecular mechanisms of differences in viability between lamb and ewe oocytes remain unknown. In the present research, we applied iTRAQ coupled with LC-MS/MS proteomic analysis in order to investigate the proteomic expression profile of granulosa cells from lambs and ewes following stimulation with FSH. We detected 5649 proteins; 574 were differentially expressed between adults and juveniles. Based on Gene Ontology enrichment and KEGG pathway analysis, the majority of DEPs are participated in metabolic processes, ribosome and MAPK signaling pathways. Expression levels in ewes turned out to be lower than lambs. Protein interaction network analysis generated by STRING identified MAPK1, SMAD2, SMAD4, CDK1, FOS and ATM as the major findings among 54 significant differentially expressed of proteins. Quantitative real-time PCR analysis was applied to verify the proteomic analysis. These proteins which were identified in lambs may contribute to the reduction of oocyte quality compared to adults. The present research provides understanding of the molecular mechanism for follicle development in lambs.

摘要

经促卵泡素(FSH)处理后,从羔羊获得的卵母细胞数量远多于成年母羊。然而,与成年母羊的卵母细胞相比,这些卵母细胞的活力通常较低。然而,羔羊和母羊卵母细胞活力差异的分子机制尚不清楚。在本研究中,我们应用同位素标记相对和绝对定量(iTRAQ)结合液相色谱-串联质谱(LC-MS/MS)蛋白质组学分析,以研究FSH刺激后羔羊和母羊颗粒细胞的蛋白质组表达谱。我们检测到5649种蛋白质;其中574种在成年羊和幼年羊之间存在差异表达。基于基因本体论(Gene Ontology)富集分析和京都基因与基因组百科全书(KEGG)通路分析,大多数差异表达蛋白参与代谢过程、核糖体和丝裂原活化蛋白激酶(MAPK)信号通路。母羊中的表达水平低于羔羊。通过STRING生成的蛋白质相互作用网络分析确定,在54种显著差异表达的蛋白质中,MAPK1、SMAD2、SMAD4、CDK1、FOS和ATM是主要发现。应用定量实时PCR分析来验证蛋白质组学分析结果。与成年羊相比,在羔羊中鉴定出的这些蛋白质可能导致卵母细胞质量下降。本研究有助于了解羔羊卵泡发育的分子机制。

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