不同发情阶段水牛唾液的DIA定量蛋白组学分析

doi:10.11843/j.issn.0366-6964.2024.11.024

摘要/Abstract

摘要：

旨在利用高通量DIA定量蛋白组学分析不同发情阶段水牛唾液中蛋白的差异。本研究将15头体况相近(平均体重为650~750 kg)，发情正常，健康状态良好的2~5胎次的尼里-拉菲母水牛作为研究对象，在相同的管理条件下，对其注射氯前列醇并计为第0天，分别在发情前期(第1天)、发情期(第3天)、发情后期(第6天)早上饲喂之前(08：00-11：00)进行唾液的采集，唾液样本每个发情时期分组相同，每3头牛分为一组，共分为5组，样品处理后进行DIA定量蛋白组学检测。结果显示，不同发情阶段水牛的唾液中共定量到1 982个蛋白质，分别在发情期/发情前期和发情期/发情后期比较组中鉴定到59和38个差异丰度蛋白。功能分析发现，CBL、SOD1、SPP1和ARPC1B等蛋白质可能在细胞调控和抗氧化、维持细胞间相互作用等方面发挥作用。KEGG通路分析显示，发情前期、发情期和发情后期水牛唾液差异丰度蛋白被富集到mTOR信号通路、孕酮介导的卵母细胞成熟等通路上。综上，通过不同发情阶段的对比，获得与繁殖相关的差异蛋白4个，CBL、ARPC1B、SOD1和SPP1。这些唾液中的差异丰度蛋白可能作为水牛发情生物标志物的关键蛋白，进而为开发水牛发情检测诊断试剂盒提供数据支撑。

关键词: 水牛, 发情鉴定, 唾液, 蛋白组学

Abstract:

This study aimed to analyze the differences in proteins in the saliva of buffaloes at different estrous stages using data-independent acquisition (DIA) proteomics. Fifteen Nili-Ravi buffaloes of similar body condition (average weight of 650-750 kg), in normal oestrus and in good health condition with 2-5 lactations were injected with chloroprostenol on day 0 of the experiment. Saliva samples were collected during proestrus (day 1), estrus (day 3), and metestrus (day 6) before morning feeding (08:00-11:00 am) under the same management conditions and subjected to data-independent acquisition proteomics analysis. Saliva samples were grouped identically for each estrous period, with every 3 cows divided into one group and a total of 5 groups. The results showed that a total of 1 982 proteins were quantified in the saliva of buffaloes at different oestrus stages. Additionally, 59 differentially abundant proteins were identified in the estrus vs proestrus comparison group, and 38 differentially abundant proteins were identified in the estrus vs metestrus comparison group. Functional analyses revealed that proteins such as CBL, SOD1, SPP1, and ARPC1B might play roles in cellular regulation, antioxidant functions, and maintenance of intercellular interactions. KEGG pathway analyses revealed that during proestrus, estrus and metestrus buffalo salivary differential abundance proteins were enriched into the mTOR signalling pathway, progesterone-mediated oocyte maturation and other pathways. In conclusion, significant differentially abundant proteins were present in the saliva of buffaloes at different estrous stages. Additionally, 4 differential proteins associated with reproduction, CBL, ARPC1B, SOD1 and SPP1, were obtained by comparing different oestrus phases. These differential proteins in saliva may serve as key proteins for estrus biomarkers in buffaloes, potentially providing a reference for the development of estrus detection diagnostic kits.

Key words: Bubalus bubalis, estrus detection, salivary, proteomics

中图分类号:

S823.83

余洲, 杨柏高, 李崇阳, 张培培, 曹建华, 牛一凡, 覃广胜, 赵学明. 不同发情阶段水牛唾液的DIA定量蛋白组学分析[J]. 畜牧兽医学报, 2024, 55(11): 5072-5084.

Zhou YU, Baigao YANG, Chongyang LI, Peipei ZHANG, Jianhua CAO, Yifan NIU, Guangsheng QIN, Xueming ZHAO. DIA Quantitative Proteomics Analysis of Buffalo Saliva at Different Estrus Stages[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5072-5084.

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名称缩写 Abbreviation	名称或注释 Name or annotation	BT vs. AT组变化倍数 Fold change	BT vs. CT组变化倍数 Fold change	BT vs. AT组 P值 P value	BT vs. CT组 P值 P value
CBL	E3泛素蛋白连接酶CBL亚型X1	+3.81	+2.29	0.009	0.04
ARPC1B	肌动蛋白相关蛋白2/3复合物亚基1B	+2.11	+1.92	0.02	0.04
SOD1	超氧化物歧化酶[Cu-Zn]	+0.53	+0.97	0.02	0.87
SPP1	骨桥蛋白	+1.65	+0.37	0.03	0.11

MapID	通路 Pathway	与该通路相关的差异蛋白的数目 Number of differential proteins associated with the pathway	P值 P value	代表性蛋白质 Typical protein
map04914	孕酮介导的卵母细胞成熟	2	0.03	RPS6KA3、KRAS
map04012	ErbB信号通路	2	0.07	CBL、KRAS
map04371	Apelin信号通路	2	0.09	SPP1、KRAS
map04919	甲状腺激素信号途径	2	0.06	KRAS
map04150	mTOR信号通路	4	0.003	RPS6KA3、KRAS

MapID	通路 Pathway	与该通路相关的差异蛋白的数目 Number of differential proteins associated with the pathway	P值 P value	代表性蛋白质 Typical protein
map00270	半胱氨酸和蛋氨酸代谢	2	0.04	LDHB、SMS
map04910	胰岛素信号通路	2	0.09	CBL

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