Plasma Proteomic Profiling Reveals the Regulatory Factors of Milk Protein Synthesis in Holstein Cows

SIMPLE SUMMARY: We aimed to determine the plasma bio-markers of cows with high long-term milk protein concentration and investigate the mechanism of plasma proteins in milk protein synthesis. Some plasma proteins are considered to be related to milk protein synthesis. However, the characteristics of these proteins with different long-term milk protein concentrations are not fully elucidated. In this study, we collected milk protein concentration data of Holstein cows for 10 months on a commercial farm. Three groups of cows (n = 10 per group) with low, medium, and high milk protein concentrations were selected. We found that cows with high milk protein concentration had higher glucose, insulin-like growth factor 1, prolactin, insulin, and growth hormone concentrations than cows with low milk protein concentration. A total of 91 differentially expressed proteins were identified between cows with high and low milk protein concentrations by plasma proteomic. Furthermore, β-casein level and mammalian rapamycin target protein pathway in bovine mammary epithelial cells were enhanced by insulin-like growth factor 1 treatment. Our findings revealed that the differences in plasma proteins in cows with different milk protein concentrations and determined that β-casein synthesis was increased via the mammalian rapamycin target protein pathway following IGF-1 treatment in vitro. ABSTRACT: Milk protein concentrations in dairy cows are considered to be related to some plasma biomolecules. However, the characteristics of plasma biomolecules in dairy cows with different long-term milk protein concentrations are not fully elucidated. This study was conducted to understand the mechanism of plasma proteins in milk protein synthesis by the comparative analysis of the plasma proteomics of cows with different milk protein concentrations. Three groups of Holstein cows (per group = 10) with low (LMP), medium (MMP), and high long-term milk protein concentrations (HMP) were selected for the experiment to determine plasma hormones, biochemical parameters, and proteome. We found that HMP cows had higher concentrations of plasma insulin-like growth factor 1 (IGF-1), glucose, prolactin, insulin, and growth hormone than LMP cows. Additionally, plasma proteomic identified 91 differential proteins, including IGF-1 between the LMP and HMP groups, and the mTOR pathway was enriched. In vitro, IGF-1 treatment increased β-casein expression and simultaneously activated S6K1 and mTOR phosphorylation in bovine mammary epithelial cells. Taken together, these data demonstrate the differences in plasma hormones, biochemical parameters, and proteome of cows with different milk protein concentrations and indicate that IGF-1 enhanced milk protein synthesis, associated with activation of the mTOR signaling pathway.