The Screening of Ovarian Candidate Proteomics Biomarkers of Dolang Sheep at Different Physiological Periods Based on iTRAQ Technology

Zilong Cui,Qianqian Chang,Weihua Chang,Mirenisa Tuersuntuoheti,Xiaojiao Yang

doi:10.5376/amb.2021.11.0001

Abstract

To screen the differentially expressed ovary proteins in Dolang sheep with oestrum, anestrus and pregnancy using isobaric tag for relative and absolute quantitation (iTRAQ) technology, which will provide the basis for the research of regulation and identification of oestrum, the diagnosis of early pregnancy. In total, we identified 4847 proteins, 470 of which were significantly different proteins. Compared with anestrus, 18 proteins were up and 102 proteins were down in oestrum, 20 proteins were up and 60 proteins were down in pregnancy; Compared with oestrum, 50 proteins were up and 24 proteins were down during pregnancy. Such as cyclin-Y isoformX1, cleavages timulation factor subunit 1 isoform X1, follistatin-related protein 1 isoform X2, nicotinate phosphoribosyl transferase isoform X1 at up-regulated, hemopexin isoform X1, apolipoprotein A-II, nucleophosmin at down-regulated, which were related to hormones generation, regulation of oestrum, zygote production and apoptosis, early embryo attachment and embryo development. Of which, cyclin-Y isoform X1 and nicotinate phosphoribosyl transferase isoform X1 significant increase in pregnancy, compared with oestrum and anestrus. The study identified different proteins at oestrum and pregnancy of Dolang sheep based on iTRAQ proteomics technology, which provide new thoughts and methods to further exploring the candidate biomarkers of sheep estrous identification and diagnosis of early gestation.

Highlights

Dolang sheep can be estrus all the year round, and it is less affected by the law of long and short sunshine
1.1 SDS PAGE analysis of ovarian protein The ovarian proteins in estrus, estrus and pregnancy were extracted by ultrasonic fragmentation method, and the protein integrity was detected by SDS-PAGE
The research of sheep reproductive diseases and reproductive physiology has developed from the initial study of a small number of major genes to the level of miRNA, transcriptomics, metabonomics and proteomics, which are biomarkers, it provides a new way of thinking for the screening and research

Summary

Introduction

Dolang sheep can be estrus all the year round, and it is less affected by the law of long and short sunshine. ITRAQ technology has been reported more and more in the regulation mechanism of animal physiological functional proteins and the screening of candidate biomarkers (Zhang et al, 2018; Wang et al, 2019). It has not been reported in the screening and identification of ovarian related proteins in different physiological periods of Dolang sheep. We used iTRAQ technology to explore the differentially expressed proteins in ovarian tissues of Dolang sheep in different physiological periods, which provided the basis for the study of the regulation mechanism of perennial estrus protein and the identification of estrus, early pregnancy examination and screening of biomarkers for disease diagnosis

Methods

Results

Conclusion