We determined the effect of insulin on milk fatty acid (FA) and lipid composition in goats. Four dairy goats, 150 d in milk, were subjected to hyperinsulinemic clamp (treatment) or saline (control) infusion for 4d in a crossover design study. Composition and concentration of plasma and milk FA, triglycerides, phospholipids, sphingolipids, and cholesterol were determined. Mammary gland biopsies were taken at the end of each experimental period and lipogenic gene expression was determined. Plasma insulin was elevated 3.5-fold, whereas plasma glucose remained constant during the treatment period. Feed intake decreased by 26% and fat yield decreased by 17% relative to controls. No change in nonesterified FA concentration was found between controls and treatment. Compared with controls, insulin decreased yield of long-chain saturated FA by 14%. Milk concentration of long-chain FA was reduced by 3%, whereas that of medium-chain FA increased by 5% during the treatment compared with controls. Hyperinsulinemic clamps increased the yields of milk phospholipids by 9% and cholesterol by 16%, whereas it only tended to decrease triglyceride yields (by 11%). Hyperinsulinemic treatment resulted in compositional changes in the milk fat globule membrane, as reflected by 15 and 9% decreases in phosphatidylethanolamine and phosphatidylcholine concentrations, respectively. Lipogenic gene expression of acyl coenzyme A carboxylase, stearoyl coenzyme A desaturase, and FA synthase did not change, whereas lipoprotein lipase gene expression tended toward an increase in the treatment period compared with controls. Hyperinsulinemic clamps reduce the availability of long-chain FA, which are considered to originate from the diet and adipose lipolysis for milk lipid synthesis by the mammary gland of goats. Under these conditions, long-chain FA might be preferentially channeled to phospholipid rather than triglyceride synthesis, hence increasing phospholipid yields. Mechanisms determining FA distribution among milk lipid components and the consequences of altered milk fat globule membrane lipid composition remained to be elucidated.

Phosphoenolpyruvate carboxykinase (PEPCK) isoforms (c, cytosolic; m, mitochondria) are expressed in the liver and mammary gland. PEPCK-c is a rate-controlling enzyme for gluconeogenesis and glyceroneogenesis; its activity is decreased by insulin. PEPCK-m expression is constitutive and functions to channel lactate toward gluconeogenesis. We hypothesized that the increase in milk protein but decrease in milk lactose and fat when a hyperinsulinemic-euglycemic clamp (HIEC) is applied to dairy goats is due to decreased expression of mammary PEPCK-c mRNA. Late lactation goats (n = 4; 150 ± 30 days in milk) were subjected to saline infusion and HIEC (104 µg insulin/h) for 4-day periods in a 2 × 2 crossover design. On day 4 of each period, a mammary biopsy (~1 g) was taken from an udder half to determine expression of PEPCK-m and PEPCK-c mRNA by real-time RT-PCR. Plasma insulin increased 3.5-fold (P< 0.002) due to the HIEC and euglycemia was maintained. The HIEC decreased (P< 0.0001) dry matter intake (28%) and milk yield (26%). While milk fat content was not affected, HIEC increased (P< 0.001) milk protein content (2.82% vs. 3.09%) but decreased (P< 0.001) milk lactose content (4.22% vs. 4.03%). Expression of PEPCK-m mRNA was 9-fold higher (P< 0.004) than that of PEPCK-c. The HIEC decreased (P< 0.03) PEPCK-c mRNA 7-fold but tended to increase (P< 0.236) PEPCK-m mRNA 1.3-fold. These results demonstrate that insulin regulates mRNA expression of mammary PEPCK isoforms, and this may underlie the changes in milk-component synthesis observed when a HIEC is applied.