Abstract
High temperature is one of the key environmental stressors affecting broiler production efficiency and meat yield. Knowledge of broiler self-regulation mechanisms under heat stress is important for the modern scale of poultry breeding. In the present study, the SWATH strategy was employed to investigate the temporal response of the broiler liver to heat stress. A total of 4,271 proteins were identified and used to generate a reference library for SWATH analysis. During this analysis, 2,377 proteins were quantified, with a coefficient of variation ≤25% among technical and biological replicates. A total of 257 proteins showed differential expression between the control and heat stressed groups. Consistent results for 26 and 5 differential proteins were validated respectively by MRM and western blotting quantitative analyses. Bioinformatics analysis suggests that the up- and down-regulation of these proteins appear involved in the following three categories of cellular pathways and metabolisms: 1) inhibit the ERK signaling pathway; 2) affect broiler liver lipid and amino acid metabolism; 3) induce liver cell immune responses to adapt to the high temperatures and reduce mortality. The study reported here provides an insight into broiler self-regulation mechanisms and shed light on the improved broiler adaptability to high-temperature environments.
Highlights
High temperature is one of the key environmental stressors affecting broiler production efficiency and meat yield
SWATH 2.0 label-free proteomics quantitative technology was utilized to obtain a global view of the proteome dynamics and changes associated with the heat response of broiler livers
The R2 values for the biological triplicates in the variance analysis were above 0.98 (Supplemental Figure 2). These analyses showed that the quantitative proteomics data are highly reproducible and reliable, while the subsequent MRM and Western blotting validation analyses confirmed the reliability of these quantitative results
Summary
High temperature is one of the key environmental stressors affecting broiler production efficiency and meat yield. Coble et al used RNA-seq technology for analysis of the transcriptome of the broiler liver under high ambient temperatures and found that high temperatures induced various physiological responses such as decreased internal temperatures, reduced hyperthermia, and cellular reactions promoting apoptosis, tissue repair, and regulating perturbed cellular calcium levels[1]. These studies show that animal adaptations to heat stress apparently depend on activation of the hypothalamic-pituitary-adrenal axis and the orthosympathetic nervous system as well as the expression of numerous stress-related genes. This new strategy is able to quantify thousands of proteins in a single measurement; the data are acquired on a fast, high-resolution Q-TOF instrument by repeatedly cycling through sequential isolation windows over the whole chromatographic elution range[22,26,27]
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