In a crossover design 10 young healthy adults (20-35 y) will receive stable isotope tracer infusions and perform a single bout of resistance exercise. Immediately after exercise participants will ingest either 3.5 oz of Salmon fillet or its constituent macronutrients as isolated amino acids and fat. Repeated blood and breath samples as well as muscle biopsies will be collected to determine whole body amino acid kinetics, muscle amino acid transporters, anabolic signalling and myofibrillar protein synthesis rates during the trials
On both infusion trials, participants will report to the laboratory at 0700 h after an overnight fast. Upon arrival to the lab, the participant will fill out a questionnaire and a baseline breath sample will be collected to determine 13CO2 enrichment by isotope ratio mass spectrometry. A Teflon catheter will be inserted into the antecubital vein for a baseline blood sample (t=-210) and then participants will receive priming doses of NaH13CO2 (2.35 µmol·kg-1), L-\[1-13C\]leucine (7.6 µmol·kg-1), and L-\[ring-2H5\]phenylalanine (2.0 µmol·kg-1). Subsequently, a continuous intravenous solution of L-\[1-13C\]leucine (0.10 µmol·kg-1·min-1) and L-\[ring-2H5\]phenylalanine (0.05 µmol·kg-1·min-1) will be initiated (t=-210) and maintained over the infusion trials. A second Teflon catheter will be inserted into a heated dorsal vein for repeated arterialized blood sampling and remained patent by a 0.9% saline drip. Breath samples and arterialized blood samples will be collected every 30 to 60 minutes during the postabsorptive and postprandial states. In the post-absorptive state of infusion trial 1, muscle biopsies will be collected at t=-150 and -30 min of infusion to determine basal-state myofibrillar protein synthesis rates, relative skeletal muscle amino acid transporter content, and anabolic-related signaling. In the subsequent cross-over trial only 1 muscle biopsy will be collected at t=-30 for Western blot analysis and postabsorptive myofibrillar protein-bound tracer enrichment. After collection of the resting muscle biopsy at t=-30 for both trials, the participants will perform resistance exercise that consists of 4 sets of 10-12 repetitions or to muscular failure at 65-70% of 1-RM for both leg press and leg extension exercise. Immediately after the completion of the exercise bout, participants will ingest either 3.5 oz of Salmon fillet or a matched isolated amino acid and fat mixture (t=0). Completion of the meal will mark the start of the postprandial stage and additional muscle biopsies will be collected at t=120 and t=300.
Study Type
INTERVENTIONAL
Participants will perform leg press and leg extension immediately prior to ingestion of salmon or isolated mixture
Participants will ingest salmon immediately after resistance exercise
Participants will ingest a mixture of isolated amino acids and fatty acids immediately after resistance exercise
Freer Hall
Urbana, Illinois, United States
Fractional synthetic rate of myofibrillar proteins
Myofibrillar protein synthesis rates will be assessed during the postabsorptive period for 3 hr and during the 5 hr after the ingestion of the experimental interventions. This will allow us to assess the change from the postabsorptive to the postprandial period.
Time frame: Postabsorptive for 3 hours, postprandial for 5 hours.
Phosphorylation of muscle anabolic signaling
Phosphorylation of anabolic signaling pathways will be assessed in the fasted state and at 2 and 5 hr after the ingestion of the experimental interventions.
Time frame: Baseline and at 2 and 5 hours after protein ingestion
Exogenous rate of leucine appearance
Leucine oxidation rates will be assessed in the fasted state and at multiple time points in the postprandial phase.
Time frame: 5 hour postprandial phase
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Allocation
RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
NONE
Enrollment
10