Milk-derived mammary epithelial cells as non-invasive source to define stage-specific abundance of milk protein and fat synthesis transcripts in native Sahiwal cows and Murrah buffaloes

The molecular physiology of milk production of two important dairy species; Sahiwal cows (Bos indicus) and Murrah buffaloes (Bubalus bubalis) are not fully understood due to constraints in obtaining mammary tissue samples because of sacred and ethical reasons. The present study suggests the use of milk-derived mammary epithelial cells (MECs) as a non-invasive method to understand molecular aspects of lactation biology in dairy animals. A total of 76 MECs were collected from five different lactation periods viz. colostrum (0–2), early (5–20), peak (30–50), mid (90–140) and late lactation (> 215 days) stages from Sahiwal cows and Murrah buffaloes to study the transcription kinetics of milk protein, fat synthesis, and their regulatory genes. Significant changes were observed in milk composition of both dairy species with lactation stages. High mRNA abundance of all milk protein and fat synthesis genes was observed in MECs of Murrah buffaloes as compared to Sahiwal cows. The mRNA abundance of caseins (CSN1S1, CSN1S2, CSN2, and CSN3) and whey protein (LALBA, LF) were higher in early lactation stage. Similarly, the expression of milk fat synthesis genes (SCD, BTN1A1, ACACA, GPAM, FAPB3, FASN) was also high in early lactation stage. The relative abundance of 4 regulatory genes (JAK2, STAT5, SREBF1 and EIF4BP41) remained high during early lactation indicating their regulatory roles in lactogenesis process. Overall, results suggested a significant effect of lactation stages on milk composition and transcription abundance of milk protein and fat synthesis genes. The present study establishes the fact that milk-derived MECs could be utilized as a valuable source to understand mammary gland functioning of native cows and buffaloes.