Leucine Supplementation Does Not Restore Diminished Skeletal Muscle Satellite Cell Abundance and Myonuclear Accretion When Protein Intake Is Limiting in Neonatal Pigs.

BACKGROUND: Rapid growth of skeletal muscle in the neonate requires the coordination of protein deposition and myonuclear accretion. During this developmental stage, muscle protein synthesis is highly sensitive to amino acid supply, especially Leu, but we do not know if this is true for satellite cells, the source of muscle fiber myonuclei. OBJECTIVE: We examined whether dietary protein restriction reduces myonuclear accretion in the neonatal pig, and if any reduction in myonuclear accretion is mitigated by restoring Leu intake. METHODS: Neonatal pigs (1.53 ± 0.2 kg) were fitted with jugular vein and gastric catheters and fed 1 of 3 isoenergetic milk replacers every 4 h for 21 d: high protein [HP; 22.5 g protein/(kg/d); n= 8]; restricted protein [RP; 11.2 g protein/(kg/d); n= 10]; or restricted protein with Leu [RPL; 12.0 g protein/(kg/d); n= 10]. Pigs were administered 5-bromo-2'-deoxyuridine (BrdU; 15 mg/kg) intravenously every 12 h from days 6 to 8. Blood was sampled on days 6 and 21 to measure plasma Leu concentrations. On day 21, pigs were killed and the longissimus dorsi (LD) muscle was collected to measure cell morphometry, satellite cell abundance, myonuclear accretion, and insulin-like growth factor (IGF) system expression. RESULTS: Compared with HP pigs, postprandial plasma Leu concentration in RP pigs was 37% and 47% lower on days 6 and 21, respectively (P < 0.05); Leu supplementation in RPL pigs restored postprandial Leu to HP concentrations. Dietary protein restriction reduced LD myofiber cross-sectional area by 21%, satellite cell abundance by 35%, and BrdU+ myonuclear abundance by 25% (P < 0.05); Leu did not reverse these outcomes. Dietary protein restriction reduced LD muscle IGF2 expression by 60%, but not IGF1 or IGF1R expression (P < 0.05); Leu did not rescue IGF2 expression. CONCLUSIONS: Satellite cell abundance and myonuclear accretion in neonatal pigs are compromised when dietary protein intake is restricted and are not restored with Leu supplementation.

Effects of dietary leucine supplementation and immune system stimulation on plasma AA concentrations and tissue protein synthesis in starter pigs.

Immune system stimulation (ISS) adversely affects protein and AA metabolism and reduces productivity in pigs. Leucine (Leu) has a regulatory role in skeletal muscle protein turnover, which may be affected by ISS. The objective of this study was to evaluate the effects of ISS and dietary Leu supplementation on the protein fractional synthesis rate (FSR) of various tissues in pigs. Yorkshire barrows were surgically fitted with jugular vein catheters and assigned to one of three dietary treatments: (i) CON, 1.36% standardized ileal digestible (SID) Leu; (ii) LEU-M, 2.04% SID Leu; and (iii) LEU-H, 2.72% SID Leu. The diets were formulated to contain all essential AA 10% above estimated requirements for maximum whole-body protein deposition for this BW range. At the start of the 36-h challenge period (initial BW = 14.5 ± 0.8 kg), ISS was induced in pigs with lipopolysaccharide (ISS+; n = 7, 8, and 7 for CON, LEU-M, and LEU-H pigs, respectively); a subset of CON pigs was injected with sterile saline (ISS-; n = 6). During challenge period, pigs were fed every 4 h and feed intake of ISS- pigs was kept equal to ISS+ pigs. At the end of the challenge period, FSR of liver, plasma, gastrocnemius, and LD proteins were determined with a flooding dose of l-[ring-2H5]phenylalanine (40 mol%). All essential AA, most nonessential AA, and plasma urea-N peaked at 12 h and declined to baseline levels at 36 h after ISS was induced in ISS+ pigs (P < 0.05), whereas plasma AA and urea-N concentrations were constant in ISS- pigs. At 36 h, dietary Leu supplementation resulted in a linear decline in plasma isoleucine, valine, glutamine, and urea nitrogen concentrations (P < 0.05), whereas plasma Leu concentration was unaffected. Liver protein FSR was increased in ISS+ pigs (P < 0.05), whereas plasma and skeletal muscle protein FSR was not affected by ISS. Dietary Leu supplementation tended to diminish liver protein FSR (linear reduction; P = 0.052) and increase gastrocnemius protein FSR (linear increase; P = 0.085) in ISS+ pigs. Leucine supplementation above estimated requirements may support repartitioning of AA from visceral to peripheral protein deposition during ISS.

Whole-body nitrogen utilization and tissue protein and casein synthesis in lactating primiparous sows fed low- and high-protein diets.

Twenty-eight lactating Yorkshire and Yorkshire × Landrace primiparous sows were used to test the hypothesis that feeding a diet with reduced CP concentration and supplemented with crystalline AA (CAA) does not decrease milk protein yield and litter growth but improves apparent N utilization for milk protein production. Sows were assigned to 1 of 2 dietary treatments: 1) control (CON; 16.2% CP; analyzed content) or 2) low CP with CAA to meet estimated requirements of limiting AA (LCP; 12.7% CP) over a 17-d lactation period. A N balance was conducted for each sow between days 13 and 17 of lactation. On day 17, a 12-h primed continuous infusion of l-[ring-2H5]-Phe was conducted on 12 sows (n = 6) with serial blood and milk sampling to determine plasma AA concentrations and Phe enrichment, and milk casein synthesis, respectively. Thereafter, sows were sacrificed and tissues were collected to determine tissue protein fractional synthesis rates (FSR). Litter growth rate and milk composition did not differ. Sows fed the LCP diet had reduced N intake (122.7 vs. 153.2 g/d; P < 0.001) and maternal N retention (13.5 vs. 24.6 g/d; P < 0.05) and greater apparent efficiency of using dietary N intake for milk production (85.1% vs. 67.5%; P < 0.001). On day 17 of lactation, all plasma essential AA concentrations exhibited a quartic relationship over time relative to consumption of a meal, where peaks occurred at approximately 1- and 4-h postprandial (P < 0.05). Protein FSR in liver, LM, gastrocnemius muscle, mammary gland, and in milk caseins did not differ between treatments. Feeding primiparous sows with a diet containing 12.7% CP and supplemented with CAA to meet the limiting AA requirements did not reduce milk protein yield or piglet growth rate and increased the apparent utilization of dietary N, Arg, Leu, Phe+Tyr, and Trp for milk protein production. The improved apparent utilization of N and AA appears to be related exclusively to a reduction in N and AA intake.

Dietary Leucine Supplementation Decreases Whole-Body Protein Turnover before, but Not during, Immune System Stimulation in Pigs.

BACKGROUND: Immune system stimulation (ISS) adversely affects protein metabolism and reduces pig productivity. Leu has a regulatory role in skeletal muscle and whole-body protein turnover, which may be affected by ISS. OBJECTIVE: We sought to determine the effect of supplemental Leu intake on whole-body protein turnover in pigs before and during ISS. METHODS: Pigs [mean ± SD initial body weight (BW): 10.6 ± 1.1 kg] were surgically fitted with jugular vein catheters and assigned to 1 of 3 treatments: 1.36% standardized ileal-digestible (SID) Leu (CON; n = 13); 2.04% SID Leu (LEU-M; n = 8); and 2.72% SID Leu (LEU-H; n = 7). Pigs were infused continuously with 0.66 ± 0.05 mmol 15N ⋅ kg BW-1 ⋅ d-1 to determine whole-body protein kinetics. The study consisted of a 72-h prechallenge period followed by a 36-h challenge period. At the start of the challenge period, ISS was induced in all LEU-M and LEU-H pigs and half of the CON pigs with LPS (ISS+); the remaining CON pigs were administered saline (ISS-). RESULTS: Whole-body protein synthesis (309, 273, and 260 ± 14 mmol N ⋅ kg BW-1 ⋅ d-1 for CON, LEU-M, and LEU-H pigs, respectively) and protein degradation (233, 203, and 185 ± 14 mmol N ⋅ kg BW-1 ⋅ d-1 for CON, LEU-M, and LEU-H pigs, respectively) were reduced with increasing Leu intake during the prechallenge period (P < 0.05). ISS reduced whole-body protein synthesis (203 compared with 169 ± 12 mmol N ⋅ kg BW-1 ⋅ d-1 for ISS- and ISS+ pigs fed CON, respectively; P < 0.05) and protein deposition (PD) (64.9 compared with 45.0 ± 2.9 mmol N ⋅ kg BW-1 ⋅ d-1 for ISS- and ISS+ pigs fed CON, respectively; P < 0.01), whereas ISS did not affect whole-body protein degradation. Leu intake did not affect whole-body protein synthesis or degradation in ISS+ pigs. CONCLUSIONS: Our results indicate that supplemental Leu intake improves the efficiency of PD rather than PD directly in healthy pigs but did not affect whole-body protein turnover during ISS.