Abstract
We compared immediate post-exercise whey protein (WP, 500 mg) versus l-leucine (LEU, 54 mg) feedings on skeletal muscle protein synthesis (MPS) mechanisms and ribosome biogenesis markers 3 h following unilateral plantarflexor resistance exercise in male, Wistar rats (~250 g). Additionally, in vitro experiments were performed on differentiated C2C12 myotubes to compare nutrient (i.e., WP, LEU) and ‘exercise-like’ treatments (i.e., caffeine, hydrogen peroxide, and AICAR) on ribosome biogenesis markers. LEU and WP significantly increased phosphorylated-rpS6 (Ser235/236) in the exercised (EX) leg 2.4-fold (P < 0.01) and 2.7-fold (P < 0.001) compared to the non-EX leg, respectively, whereas vehicle-fed control (CTL) did not (+65 %, P > 0.05). Compared to the non-EX leg, MPS levels increased 32 % and 52 % in the EX leg of CTL (P < 0.01) and WP rats (P < 0.001), respectively, but not in LEU rats (+15 %, P > 0.05). Several genes associated with ribosome biogenesis robustly increased in the EX versus non-EX legs of all treatments; specifically, c-Myc mRNA, Nop56 mRNA, Bop1 mRNA, Ncl mRNA, Npm1 mRNA, Fb1 mRNA, and Xpo-5 mRNA. However, only LEU significantly increased 45S pre-rRNA levels in the EX leg (63 %, P < 0.001). In vitro findings confirmed that ‘exercise-like’ treatments similarly altered markers of ribosome biogenesis, but only LEU increased 47S pre-rRNA levels (P < 0.01). Collectively, our data suggests that resistance exercise, as well as ‘exercise-like’ signals in vitro, acutely increase the expression of genes associated with ribosome biogenesis independent of nutrient provision. Moreover, while EX with or without WP appears superior for enhancing translational efficiency (i.e., increasing MPS per unit of RNA), LEU administration (or co-administration) may further enhance ribosome biogenesis over prolonged periods with resistance exercise.
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Acknowledgments
We graciously thank Dr. Troy Horberger for his insight into the SUnSET technique as well as Dr. John McCarthy and Tyler Kirby for their intellectual insight on ribosome biogenesis. Funding from MusclePharm Research Institute was used to fund the direct costs of this study, undergraduate technical support, and publication costs of these data.
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Besides M.K. and J.R.M., none of the authors have non-financial and/or financial competing interests. M.K. and J.R.M. are employed by the MusclePharm Research Institute, but both intellectually contributed to study design and data write-up. Therefore, all co-authors agreed that their intellectual input into this project warranted co-authorship.
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This article does not contain any studies with human participants performed by any of the authors. All of the proposed animal studies were approved by the Institutional Animal Care and Use Committee at Auburn University. When applicable, replacement of rats with cell culture experiments, a reduction in the number of rats used, and refinement of methods to alleviate animal discomfort were used in the animal protocol.
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Mobley, C.B., Fox, C.D., Thompson, R.M. et al. Comparative effects of whey protein versus l-leucine on skeletal muscle protein synthesis and markers of ribosome biogenesis following resistance exercise. Amino Acids 48, 733–750 (2016). https://doi.org/10.1007/s00726-015-2121-z
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DOI: https://doi.org/10.1007/s00726-015-2121-z