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RESEARCH ARTICLE
Contents Vol 64(7)

TMT-based quantitative proteomic analysis of serum from domestic sheep in early pregnancy

Yujun Ren https://orcid.org/0000-0003-1870-9141 A B # , Zhunxuan Wang A B # , Yishan Sun B , Hongbin Gong B , Su Xie B , Ruonan Gao B , Xin Chen B , Qingchun Li B , Shihao Lu B , Tao Huang https://orcid.org/0000-0003-2943-5131 A B * and Min Yang B *
+ Author Affiliations
- Author Affiliations

A State Key Laboratory for Sheep Genetic Improvement and Healthy Production, Xinjiang Academy of Agricultural and Reclamation Science, Xinjiang, China.

B College of Animal Science and Technology, Shihezi University, Shihezi, China.

* Correspondence to: taohuang100@sina.com, yangmindk@163.com

Handling Editor: Gry Boe-Hansen

Animal Production Science 64, AN22445 https://doi.org/10.1071/AN22445
Submitted: 2 December 2022  Accepted: 8 April 2024  Published: 1 May 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Context

Timely pregnancy diagnosis is critical for reducing non-production days and improving the reproductive efficiency of sheep flocks.

Aims

The aim of this study was to investigate the serum proteomic profile of ewes in different gestational stages and explore the potential of differentially expressed proteins as biomarkers for early gestation.

Methods

Serum samples were collected from two groups: pregnant sheep (n = 4) and non-pregnant sheep (n = 4) on Day 14 after mating. The differentially expressed proteins were detected using tandem mass-tag (TMT) labelling with liquid chromatography tandem-mass spectrometry. The differentially expressed proteins were confirmed using enzyme-linked immunosorbent assay (ELISA). Furthermore, six selected proteins were evaluated in ewes (n = 35) on Day 14 from mating by using ELISA to assess their potential as biomarkers for early pregnancy. Receiver-operating characteristic (ROC) curves were used to analyse the diagnostic accuracy of these proteins.

Key results

Twenty proteins, of a total of 555 proteins detected, showed differential expression between pregnant and non-pregnant sheep. From the ROC-curve data, CHI3L1 (AUC = 0.992), PSMB4 (AUC = 0.976) and LGALS3BP (AUC = 0.844) had high predictive value in the diagnosis of early pregnancy.

Conclusions

In this study, CHI3L1, PSMB4 and LGALS3BP were found to be differentially expressed proteins in the serum of pregnant sheep. Therefore, these three proteins carry a high probability as candidate biomarkers for early gestational embryo attachment in ewes.

Implications

Early pregnancy diagnosis enables timely identification of non-pregnant ewes, resulting in optimised resource allocation and enhanced economic benefits for farmers by reducing production costs.

Keywords: CHI3L1, early pregnancy, embryo implantation, PSMB4, reproductive, serum, sheep, tandem mass-tag labelling.

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