| 摘要: |
| 本研究构建了水牛卵巢黄体形成及退化过程的定量蛋白质表达谱，从中寻找与水牛发情周期相关的重要蛋白质。为研究水牛卵巢周期性变化的分子机制提供蛋白质水平的数据支撑，进而为提高水牛繁殖效率奠定实验基础。以水牛排卵后的红体期（corpus hemorrhagicum, CH）、黄体期（corpus luteum, CL）和白体期（corpus fibrosum, CF）的卵巢作为研究对象，利用串联质谱标签(tandem mass tag, TMT)标记结合液质联用（(liquid chromatography/mass spectrometry,LC-MS/MS）的定量蛋白质组学技术，采用相关软件对得到的差异表达蛋白质进行生物信息学分析。同时对与水牛发情周期相关的重要差异蛋白质：单核巨噬细胞分化抗原（CD14）、通用转录因子Ⅱ-Ⅰ（GTF2I）、羟基类固醇（17β）脱氢酶1（HSD17B1）、U6 snRNA相关Sm样蛋白质 (LSM1)和纤溶酶原激活物抑制物 (SERPINE1)进行了qRT-PCR分析验证。结果显示，共鉴定得到2 343种蛋白质，其中差异表达蛋白质有284种（差异倍数≥2.0），初步构建了水牛卵巢黄体形成和退化的定量蛋白质表达谱。对其中的五种重要差异蛋白质(CD14、GTF2I、HSD17B1、LSM1和SERPINE1)进行qRT-PCR验证，结果有四种（CD14、HSD17B1、LSM1和SERPINE1）的qPCR结果与质谱结果相一致，仅GTF2I与质谱结果不一致，可能是由于GTF2I存在转录后调控。本研究首次从蛋白质水平对水牛卵巢排卵后的周期性变化的分子机制进行了探究，同时为其他家畜品种发情周期的研究提供了新的研究思路。 |
| 关键词: 水牛 发情周期 黄体 定量蛋白质组学 生物信息学 |
| DOI：10.14188/j.ajsh.2018.05.002 |
| 分类号: |
| 基金项目:国家自然科学基金（31460603）；广西自然科学基金（2014GXNSFAA118134）；广西研究生教育创新计划项目（YCBZ2017005） |
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| The protein profile and bioinformatic analysis of the formation and regression of the corpus luteum in buffalo |
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CHEN Fumei,YAO Shun,CHEN Dongrong,XU Zhuangzhuang,HOU Zhen,PU Liping,ZHANG Pengfei,ZHAO Xiuling,ZHANG Ming
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State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530004, Guangxi, China
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| Abstract: |
| Buffalo is an important subtropical livestock breed in Asia, but its low productivity limits the commercialized development . The main reason is that the oestrus performance of female buffalo is not obvious, and its reproduction rate greatly depends on the cure rate of the abnormal oestrus. The estrous cycle of the female buffalo is accompanied by the formation and degeneration of the corpus luteum. In this study, we conducted the quantitative proteomics technology using tandem mass tag coupled to LC-MS/MS (Liquid chromatography/mass spectrometry) to construct the protein expression file of buffalo ovary from three different physiological stages (corpus hemorrhagicum, CH; corpus luteum, CL and corpus fibrosum, CF). The results showed that a total of 2343 proteins were identified , in which the 284 proteins were differentially expressed (fold change≥2). In addtion, the results of bioinformatics analysis showed that these differentially expressed proteins were associated with many molecular functions, such as nucleotide binding, serine-type endopeptidase inhibitor activity. Also, the differentially expressed proteins were involved in the negative regulation of activated T cell proliferation, mRNA processing, and so on. Finally, we selected five important proteins (CD14,GTF2I,HSD17B1,LSM1,SERPINE1) to make the validation using RT-qPCR. The result of RT-qPCR showed that the four genes (CD14, HSD17B1,LSM1,SERPINE1) were consistent with the result of quantitative proteomics analysis, and only the result of GTF2I was not consistent with its quantitative proteomics result, and we speculated that this protein might exist the post-transcription regulator. Our study firstly reported the investigation for the molecular mechanism about the formation and degeneration of the corpus luteum in buffalo ovary at the protein level. |
| Key words: buffalo estrous cycle corpus luteum quantitative proteomics bioinformatics |
