Acta Veterinaria et Zootechnica Sinica ›› 2024, Vol. 55 ›› Issue (11): 5072-5084.doi: 10.11843/j.issn.0366-6964.2024.11.024

• Animal Biotechnology and Reproduction • Previous Articles     Next Articles

DIA Quantitative Proteomics Analysis of Buffalo Saliva at Different Estrus Stages

Zhou YU1(), Baigao YANG1, Chongyang LI1, Peipei ZHANG1, Jianhua CAO1, Yifan NIU1, Guangsheng QIN2, Xueming ZHAO1,*()   

  1. 1. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
    2. Buffalo Research Institute of Guangxi Zhuang Nationality Autonomous Region, Chinese Academy of Agricultural Sciences, Nanning 530001, China
  • Received:2024-05-09 Online:2024-11-23 Published:2024-11-30
  • Contact: Xueming ZHAO E-mail:yz.zhouyu@qq.com;zhaoxueming@caas.cn

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

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