Glycosylphosphatidylinositol-anchored high-density lipoprotein-binding protein 1 (GPIHBP1) is among the many candidate genes for regulating milk production traits in dairy cattle that have been identified via quantitative trait locus (QTL) mapping and genome-wide association studies (GWAS). Our previous studies confirmed that a G-to-A mutation at chr14:2553998 is the main cause of GPIHBP1-related effects on milk fat content. In this study, we discovered that GPIHBP1 may be a strong candidate gene for the regulation of milk protein traits. We performed overexpression and RNAi experiments to assess GPIHBP1 in bovine primary mammary epithelial cells (BMECs) and identified mRNA expression patterns of several important milk protein-related genes using real-time quantitative PCR. After the transient transfection of BMECs with GPIHBP1, the transcription levels of casein genes (CSN1S1, CSN1S2, CSN2, and CSN3) and lactoferrin (LTF) decreased, whereas beta-lactoglobulin (LGB) expression increased. The GPIHBP1 RNAi experiment produced changes in gene expression that were completely opposite to those observed in the GPIHBP1 overexpression experiment. Furthermore, among the assessed genes, CSN3, LTF, and LGB exhibited significant changes in mRNA expression (p < 0.05). The findings of this study show that bovine GPIHBP1 is involved in the process of milk protein biosynthesis and may be considered as a functional gene for the milk protein yield trait.
Keywords: Bovine mammary epithelial cell; GPIHBP1; RNAi; milk protein traits; overexpression.