Influence of Leucine Enriched Amino Acids on Myofibrillar and Collagen Protein Synthesis

Daniel Moore8 enrolled

Overview

The primary objective will be to determine the effect of leucine-enriched essential amino acids (LEAA) compared to carbohydrate placebo on dietary incorporation of \[D5\] Phenylalanine \& \[D5\] Glycine into the three skeletal muscle protein pools (myofibrillar, sarcoplasmic and collagen), both following resistance exercise and at rest, with the two tracers provided as a 'intrinsically labeled' bolus. Other outcomes will relate to molecular regulation of protein synthesis.

Study Type

INTERVENTIONAL

Allocation

RANDOMIZED

Purpose

TREATMENT

Masking

TRIPLE

Enrollment

Conditions

Interventional

Interventions

Leucine enriched amino acidsDIETARY_SUPPLEMENT

Leucine enriched essential amino acids (4g total, of which 1.6g are leucine). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda.

Carbohydrate PlaceboDIETARY_SUPPLEMENT

iso-caloric placebo (4g maltodextrin). 20g carbohydrate, 2g of protein free drink powder, and 5g of Splenda.

Eligibility

Sex: ALLMin age: 18 YearsMax age: 35 YearsHealthy volunteers:

Medical Language ↔ Plain English

Inclusion Criteria: * Healthy, male and female, recreationally-active participants. * Healthy will be defined as screened by the PAR-Q+ (The Physical Activity Readiness Questionnaire for everyone; Appendix B). * Self-reported not having engaged in resistance exercise and/or lower body plyometrics for at least 3 months prior to the study. * Participants will be aged 18-35 years old. * Participants are willing to abide by the compliance rules of this study. * Self-reported regular menstrual cycle (25-35d) within the last 3 months (female participants). Exclusion Criteria: * Inability to adhere to any of the compliance rules judged by principle investigator or medical doctor. * Self-reported regular tobacco use. * Self-reported illicit drug use (e.g. growth hormone, testosterone, etc.). * Individuals who have participated in studies within the past year involving any of the stable isotopes in the study. * Use of birth control and discontinued use within the last 3 months (female participants).

Locations (1)

Goldring Centre for High Performance Sport at the University of Toronto

Toronto, Ontario, Canada

Outcomes

Primary Outcomes

Myofibrillar Protein synthesis

Rate of incorporation of orally ingested amino acid tracers (D5 Phe and D5 Gly) into myofibrillar proteins isolated from skeletal muscle biopsies 4h following drink (LEAA or PLA) consumption and resistance exercise or at rest. Enrichment will be quantified via tandem LC-MS/MS to derive a rate of incorporation over the 4h period.

Time frame: 4 hours

Secondary Outcomes

Sarcoplasmic protein synthesis

Rate of incorporation of orally ingested amino acid tracers (D5 Phe and D5 Gly) into sarcoplasmic (i.e., soluble) proteins isolated from skeletal muscle biopsies 4h following drink consumption (LEAA or PLA) and resistance exercise or at rest. Enrichment will be quantified via tandem LC-MS/MS.

Time frame: 4 hours

Collagen protein synthesis

Rate of incorporation of orally ingested amino acid tracers (D5 Phe and D5 Gly) into collagen (i.e., non-soluble) proteins isolated from skeletal muscle biopsies 4h following drink consumption (LEAA or PLA) and resistance exercise or at rest. Enrichment will be quantified via tandem LC-MS/MS.

Time frame: 4 hours

Protein localization via immunofluorescence

Antibody-determined Phosphorylated RPS6 (S240/244) and mTOR protein localization in human muscle cross sections and longitudinal sections as examined via immunofluorescence. Muscle sections (both longitudinal and cross-sections) will be derived from snap-frozen muscle biopsies performed 4h following drink consumption (LEAA or PLA) with or without resistance exercise (unilateral). Localization will be performed as done by Hodson et al., (2022; AJP Cell). Particularly, the localization of these targets relative to the cell periphery and z-discs (longitudinal sections only) will be analyzed via colocalization analysis.

Time frame: 4 hours

Data from ClinicalTrials.gov

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