Abstract
BackgroundTibetan pigs (TP) exhibit heritable adaptations to their hypoxic environments as a result of natural selection. However, candidate proteins that affect the sperm quality of boars on plateaus have not yet been clearly investigated.MethodsIn this study, to reveal the candidate proteins that affect the quality of spermatozoa of boars on plateaus, we analyzed the sperm quality using computer-assisted semen analysis (CASA) system and reactive oxygen species (ROS) levels. We also compared the proteomes of sperm proteomes between TP and Yorkshire pigs (YP) raised at high altitudes using the isobaric tags for relative and absolute quantitation (iTRAQ) in combination with the liquid chromatography-tandem mass spectrometry (LC–MS/MS) proteomic method, and confirmed the relative expression levels of the four proteins by western blotting.ResultsThe sperm quality of the TP was superior to that of the YP on plateaus. Of the 1,555 quantified proteins, 318 differentially expressed proteins (DEPs) were identified. Gene ontology (GO) analysis revealed that the DEPs were predominantly associated with the sorbitol metabolic process, removal of superoxide radicals, cellular response to superoxide, response to superoxide and regulation of the mitotic spindle assembly. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were mainly enriched in pathways involved in the regulation of the actin cytoskeleton, glutathione metabolism, oxidative phosphorylation, and estrogen signaling. Based on the protein–protein interaction (PPI) network analysis, we identified 8 candidate proteins (FN1, EGF, HSP90B1, CFL1, GPX4, NDUFA6, VDAC2, and CP) that might play important roles and affect the sperm quality of boars on plateaus. Moreover, the relative expression levels of four proteins (CFL1, EGF, FN1, and GPX4) were confirmed by western blot analysis.ConclusionsOur study revealed 8 candidate proteins (FN1, EGF, HSP90B1, CFL1, GPX4, NDUFA6, VDAC2, and CP) that affect the sperm quality of boar on plateaus and provide a reference for further studies on improving sperm quality and the molecular breeding of boars on plateaus.
Highlights
Tibetan pigs (TP) exhibit heritable adaptations to their hypoxic environments as a result of natural selection
Comparison of sperm quality in the TP and the Yorkshire pigs (YP) The results showed that the hypothesis of homogeneity variance was satisfactory, and the sperm
Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differentially expressed proteins (DEPs) The DEPs were classified by gene ontology annotation based on three categories: biological process (BP), molecular function (MF), and cellular component (CC) (Additional file 4: Table S4, only result for false-positive detection rate (FDR) P < 0.05 is displayed)
Summary
Tibetan pigs (TP) exhibit heritable adaptations to their hypoxic environments as a result of natural selection. Candidate proteins that affect the sperm quality of boars on plateaus have not yet been clearly investigated. The reproductive system and antioxidants involved in oxidative stress in male boars are affected by hypobaric hypoxia [1]. Tibetan pigs (TP) is a unique and. The sperm motility of TP is higher than that of YP on plateaus, but analyses of other sperm quality parameters of TP and YP raised at high altitudes have not yet been conducted. It is necessary to evaluate sperm ROS levels when performing sperm quality analysis. In-depth proteomic analyses of boar spermatozoa have been performed using shotgun and gel-based methods [6]. The sperm proteomes of TP and YP raised at high altitudes have not yet been compared. The sperm proteomes of TP and YP raised at high altitudes have not yet been compared. iTRAQ is a powerful technique for quantitatively analyzing proteomes [9] and has high sensitivity, good repeatability, the ability to label almost all enzymatic peptides, and the ability to label 8 samples simultaneously [10]
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