Hormone-sensitive lipase protein expression and extent of phosphorylation in subcutaneous and retroperitoneal adipose tissues in the periparturient dairy cow

Lipomobilization is essential for dairy cows to balance the energy requirement for milk production in early lactation. This study aimed to determine the role of hormone-sensitive lipase (HSL) and its activation by phosphorylation at Ser 660 (HSLp660) and 563 (HSLp563) in different adipose tissue depots as influenced by time and postpartum diet in dairy cows. Biopsy samples were obtained from s.c. (SCAT) and retroperitoneal (RPAT) adipose tissues of 20 Holstein cows 21 d prepartum, and 1 and 21 d postpartum. After d 1 postpartum, cows were randomly assigned to 2 groups (n = 10). Groups received diets with either a concentrate-to-roughage ratio on a dry matter basis of 30:70% (low-concentrate, LC, group) or 60:40% (high-concentrate group), fed until the third biopsy sampling 21 d postpartum. Dry matter intake, milk yield, and milk composition were recorded. Blood samples were taken weekly, starting 21 d prepartum and analyzed for nonesterified fatty acids, beta-hydroxybutyrate (BHBA), glucose, and insulin. Protein expression of HSL and its extent of phosphorylation in adipose tissue were measured by semiquantitative Western blotting. Total HSL expression was lower in both adipose tissues 1 d after calving compared with prepartum sampling (SCAT: 4.10 +/- 0.5 vs. 2.4 +/- 0.3; RPAT: 11.1 +/- 1.3 vs. 6.6 +/- 1.1). Phosphorylation at Ser 660 was higher 21 d postpartum compared with 21 d prepartum in RPAT (2.9 +/- 0.3 vs. 4.6 +/- 0.6). Phosphorylation at Ser 563 was higher 21 d postpartum than 21 d prepartum in SCAT (0.6 +/- 0.1 vs. 3.9 +/- 1.1), and in RPAT a difference was observed between 21 d prepartum and 1 d postpartum (1.0 +/- 0.1 vs. 3.3. +/- 0.6). On d 21 postpartum, the LC group showed a lower extent of Ser 563 phosphorylation in RPAT (3.9 +/- 0.8 vs. 10.0 +/- 1.9) and a higher concentration of serum BHBA (0.77 +/- 0.05 vs. 0.47 +/- 0.11) than did the high-concentrate group. An inhibitory influence of higher BHBA concentrations on HSL phosphorylation in the LC group could be a possible explanation. On comparing RPAT to SCAT, HSL expression and the extent of Ser 660 and 563 phosphorylation was higher in RPAT at 21 d prepartum (HSL: 4.1 +/- 0.5 vs. 11.1 +/- 1.2; HSLp660 1.3 +/- 0.2 vs. 2.9 +/- 0.3; HSLp563: 0.6 +/- 0.1 vs. 1.0 +/- 0.1). In conclusion, the postpartum feeding regimen influenced the phosphorylation pattern, especially in RPAT, implying a regulatory role for different phosphorylation sites in adaptive lipolysis of dairy cows. It is suggested that RPAT is more sensitive to periparturient challenges than is SCAT.