Intermittent Leucine Pulses Enhance Skeletal Muscle mTOR Signaling and Protein Synthesis in Continuously Fed Preterm Pigs

Abstract

Abstract Objectives Extrauterine growth restriction in premature infants is associated with reduced lean mass and long-term morbidities. We have reported previously that intermittent parenteral pulses of Leu promote skeletal muscle mTOR signaling and protein synthesis of continuously fed neonatal pigs born at term. The objective of this study was to determine the effect of prematurity on the response of skeletal muscle anabolic pathways to intermittent parenteral Leu pulses in continuously fed pigs. Methods Pigs delivered 10 d preterm by C-section were fitted with jugular vein and carotid artery catheters and an orogastric feeding tube. Pigs were advanced from parenteral to enteral feeding over 4 d (195 kcal · kg−1 · d−1; 13.5 g protein · kg−1 · d−1). On day 4, pigs were assigned to 1 of 4 treatments: 1) ALA (continuous feeding, 7.5 mL · kg−1 · h−1; 800 μmol Ala · kg−1 · h−1 every 4 h; n = 7); 2) L1 × (continuous feeding; 800 μmol Leu · kg−1 · h−1 every 4 h; n = 6); 3) L2 × (continuous feeding; 1600 μmol Leu · kg−1 · h−1 every 4 h; n = 6); and 4) INT (intermittent feeding; 30 mL · kg−1 fed over 15 min every 4 h; n = 5). On day 5, pigs received L-[ring-2H5]-Phe 30 min after starting the pulse (groups 1, 2, and 3) or meal feeding (group 4). Pigs were euthanized 30 min after isotope injection and longissimus dorsi muscle was collected. Protein synthesis was determined by LC/MS-MS. Indices of amino acid signaling and mTOR activation were determined by immunoprecipitation and immunoblot assays. Results Skeletal muscle protein synthesis was higher in L2 × (+37%) and INT (+31%) compared to ALA (P < 0.05), but was not different between L2 × and INT; protein synthesis in L1 × was intermediate and not different from all other groups. The phosphorylation of 4EBP1, downstream of mTOR, was higher in L2 × and INT compared to ALA (P < 0.05), whereas 4EBP1 phosphorylation in L1 × was lower compared to INT (P < 0.05) but not different compared to ALA and L2 × . The abundance of mTOR · RagA complex, upstream of mTOR and activated in response to Leu, was higher in L2 × and INT compared to ALA and L1 × (P < 0.05). Conclusions These results show that parenteral Leu can enhance anabolic signaling and protein synthesis in skeletal muscle during continuous feeding in preterm pigs, but the dose required is higher than in pigs born at term. Funding Sources Research was supported by NIH and USDA.