About this trial
The muscles of the body are constantly breaking down old proteins and building new ones. These two processes, protein breakdown and protein synthesis, together are known as protein turnover. Protein turnover is essential for maintaining healthy muscle.
Despite its importance, females have historically been underrepresented in protein metabolism research. A long-standing assumption has been that fluctuations in female sex hormones such as estrogen and progesterone, whether across the natural menstrual cycle or in individuals using oral contraceptives (OCs), make metabolism and training responses too variable to study. Because of this, many researchers have excluded female participants for logistical reasons.
Resistance exercise, such as weightlifting, is the most effective way to increase muscle size and strength. Each resistance-training session triggers muscle protein synthesis (MPS), the process by which new muscle proteins are built. Consuming dietary protein or individual amino acids further increases the rate at which new proteins are formed. Over time, higher rates of protein synthesis support muscle growth and the maintenance of other lean tissues in the body.
The purpose of this study is to examine how menstrual cycle phases and OC use influence the synthesis of proteins in both muscle tissue and the rest of the body. Improving scientific understanding in this area will support more effective, evidence-based training and nutrition recommendations for females.
Eligibility criteria
This trial accepts healthy volunteersQualifiers
Age: 18-40 years
BMI between 18.5-29.9 kg/m2 (non-obese)
Recreationally active (resistance train minimum twice a week)
Using monophasic or triphasic oral contraceptives for >1 year (for oral contraceptive users)
Disqualifiers
Mid-luteal progesterone levels <16umol
Chronic disease diagnosis (cardiovascular, thyroid, diabetes)
Current or recent remission of cancer
Regular use of non-steroidal anti-inflammatory drugs (NSAIDs; except low-dose aspirin), anticoagulants
Trial design
Crossover
Treatments tested in this trial
Protein Beverage
Dietary supplementParticipants will consume 16 half-hourly (8 hours) isoenergetic, isonitrogenous beverages containing 1.2 g/kg/d protein.
Stable Isotope Tracer [D5]phenylalanine
Dietary supplementProtein beverages will be enriched with the stable isotope \[D5\]phenylalanine. The use of \[D5\]phenylalanine will allow for determination of muscle protein synthesis.
Stable Isotope Tracer [13C]phenylalanine
Dietary supplementProtein beverages will be enriched with the stable isotope tracer \[13C\]phenylalanine. The use of \[13C\]phenylalanine will allow for the determination of the fate of amino acids in the body (incorporation into body protein or oxidation) which can be used to determine protein requirements.
Treatment groups
Trial outcomes
Primary outcomes
Muscle Protein Synthesis (MPS)
Muscle protein synthesis (MPS) will be assessed during an 8-hour post-exercise and feeding period using a stable isotope tracer, L-\[ring-D5\]phenylalanine (\[D5\]Phe). The incorporation of the \[D5\]Phe tracer into myofibrillar muscle proteins, collected from muscle biopsies (pre-exercise and post-8h feeding), will be analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine the fractional synthesis rate (FSR) of muscle proteins. This will help us understand how menstrual cycle phases and oral contraceptive use influence MPS, providing insight into optimizing protein intake and training adaptations in female athletes.
Secondary outcomes
Whole Body Protein Synthesis (WBPS)
Whole body protein synthesis (WBPS) will be measured over a 4-hour period using the Indicator Amino Acid Oxidation (IAAO) methodology. Participants will consume a stable isotope tracer, L-\[1-13C\]phenylalanine (\[13C\]Phe), and breath samples will be collected every 30 minutes during the final 4 hours of the metabolic trial. The enrichment of 13CO₂ in breath samples will be analyzed to determine phenylalanine oxidation rates, providing an estimate of whole-body protein turnover and dietary protein utilization. This will allow us to assess whether hormonal fluctuations influence protein metabolism and nutritional requirements in female athletes.
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