Expression Profiling of Glucose Transporter 1 (GLUT1) and Apoptotic Genes (BAX and BCL2) in Milk Enriched Mammary Epithelial Cells (MEC) in Riverine Buffalo during Lactation

Abstract

Lactation is an important physiological process in dairy animals. During lactation, up to 85% of the body glucose is directed toward the mammary glands for milk synthesis. Studies related to lactation physiology are generally carried out on mammary biopsies, which may adversely affect animal health. In the present study, milk enriched MEC were used to study the expression pattern of GLUT1 and apoptotic genes (BAX and BCL2) across different stages of lactation in riverine buffalo in relation to milk yield. MEC were enriched from milk using cytokeratin-8 antibodies coated magnetic beads. Total RNA isolated from enriched MEC showed significant correlation (r 2 = 0.92 ± 0.02) with the milk yield at different stages of lactation. GLUT1 expression pattern correlated with the milk yield as highest GLUT1 expression (4.68 ± 0.79) was observed during peak-lactation (90 days post-parturition), whereas low GLUT1 expression (1.01 ± 0.1, 15 d; 0.71 ± 0.03, 30 d) was observed during early lactation. The BAX/BCL2 ratio was high (1.02 ± 0.2, 15 d; 0.94 ± 0.06, 30 d) during the early phase of lactation, indicating high rate of apoptosis, whereas low BAX/BCL2 ratio (0.25 ± 0.03, 60 d; 0.42 ± 0.04, 90 d) was observed during mid-lactation coinciding with the increase in RNA concentration and milk yield. Highest BAX/BCL2 ratio (1.41 ± 0.3, 120 d; 4.02 ± 0.6, 240 d) was observed during late lactation i.e., 240 days, which was also reflected as decline in milk yield and RNA concentration. Also, BAX/BCL2 ratio in milk enriched MEC was in accordance with RNA concentration in MEC and milk yield at different phases of lactation. Our study showed that expression pattern of genes under study (GLUT1, BAX, and BCL2) in milk enriched MEC correlated well with important physiological properties such as milk yield in buffalo.