Proteome Profile and Quantitative Proteomic Analysis of Buffalo (Bubalusbubalis) Follicular Fluid during Follicle Development.
Proteome Profile and Quantitative Proteomic Analysis of Buffalo (Bubalusbubalis) Follicular Fluid during Follicle Development.
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卵泡发育过程中水牛 (Bubalusbubalis) 卵泡液的蛋白质组谱和定量蛋白质组分析
DOI：
10.3390/ijms17050618
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发表时间：
2016-04-29
影响因子：
5.6
通讯作者：
Zhang M
中科院分区：
文献类型：
--
作者：
Fu Q;Huang Y;Wang Z;Chen F;Huang D;Lu Y;Liang X;Zhang M
卵泡液（FF）积聚在卵泡腔中，为卵母细胞发育提供微环境。FF在卵泡生长和卵母细胞成熟中起重要作用。FF为研究布法罗卵泡发育过程提供了一个独特的窗口。观察到的低质量的布法罗卵母细胞可能是由于卵泡微环境差。研究布法罗FF（BFF）中发现的蛋白质应该提供对卵泡发育过程的深入了解，并提供对卵泡内成熟和卵母细胞质量的进一步了解。本文采用蛋白质组学方法对牛卵泡液蛋白质组进行了分析。SDS-PAGE分离结合质谱法用于产生蛋白质组数据集。总共鉴定了363个蛋白质，并通过基因本体论术语进行了分类。这些蛋白质被分配到153条通路，包括信号通路。为了评估具有不同卵泡大小（小，<4 mm;和大，>8 mm）的BFF之间表达的蛋白质的差异，进行基于多维液相色谱预分级串联Orbitrap质谱鉴定的定量蛋白质组学分析。11个差异表达的蛋白质（6下调和5上调大BFF）进行了鉴定，并分配到各种功能过程，包括丝氨酸蛋白酶抑制，氧化保护和补体级联系统。通过Western blotting验证了Vimentin、Peroxiredoxin-1和SERPIND 1 3个差异表达蛋白，与定量蛋白质组学结果一致。我们的数据集提供了关于BFF中存在的蛋白质的新信息，并应有助于开发新的生物标志物。这些差异表达的蛋白质阐明了卵泡微环境中蛋白质的大小依赖性变化。
Follicular fluid (FF) accumulates in the antrum of the ovarian follicle and provides the microenvironment for oocyte development. FF plays an important role in follicle growth and oocyte maturation. The FF provides a unique window to investigate the processes occurring during buffalo follicular development. The observed low quality of buffalo oocytes may arise from the poor follicular microenvironment. Investigating proteins found in buffalo FF (BFF) should provide insight into follicular development processes and provide further understanding of intra-follicular maturation and oocytes quality. Here, a proteomic-based approach was used to analyze the proteome of BFF. SDS-PAGE separation combined with mass spectrometry was used to generate the proteomic dataset. In total, 363 proteins were identified and classified by Gene Ontology terms. The proteins were assigned to 153 pathways, including signaling pathways. To evaluate difference in proteins expressed between BFF with different follicle size (small, <4 mm; and large, >8 mm), a quantitative proteomic analysis based on multi-dimensional liquid chromatography pre-fractionation tandem Orbitrap mass spectrometry identification was performed. Eleven differentially expressed proteins (six downregulated and five upregulated in large BFF) were identified and assigned to a variety of functional processes, including serine protease inhibition, oxidation protection and the complement cascade system. Three differentially expressed proteins, Vimentin, Peroxiredoxin-1 and SERPIND1, were verified by Western blotting, consistent with the quantitative proteomics results. Our datasets offers new information about proteins present in BFF and should facilitate the development of new biomarkers. These differentially expressed proteins illuminate the size-dependent protein changes in follicle microenvironment.