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Dietary leucine supplementation alters energy metabolism and induces slow-to-fast transitions in longissimus dorsi muscle of weanling piglets

  • Qiwen Fan (a1) (a2) (a3), Baisheng Long (a1) (a2) (a3), Guokai Yan (a1) (a2) (a3), Zhichang Wang (a1) (a2) (a3), Min Shi (a1) (a2) (a3), Xiaoyu Bao (a1) (a2) (a3), Jun Hu (a1) (a2) (a3), Xiuzhi Li (a1) (a2) (a3), Changqing Chen (a1) (a2) (a3), Zilong Zheng (a1) (a2) (a3) and Xianghua Yan (a1) (a2) (a3)...


Leucine plays an important role in promoting muscle protein synthesis and muscle remodelling. However, what percentage of leucine is appropriate in creep feed and what proteome profile alterations are caused by dietary leucine in the skeletal muscle of piglets remain elusive. In this case, we applied isobaric tags for relative and absolute quantitation to analyse the proteome profile of the longissimus dorsi muscles of weanling piglets fed a normal leucine diet (NL; 1·66 % leucine) and a high-leucine diet (HL; 2·1 % leucine). We identified 157 differentially expressed proteins between these two groups. Bioinformatics analysis of these proteins exhibited the suppression of oxidative phosphorylation and fatty acid β-oxidation, as well as the activation of glycolysis, in the HL group. For further confirmation, we identified that SDHB, ATP5F1, ACADM and HADHB were significantly down-regulated (P<0·01, except ATP5F1, P<0·05), whereas the glycolytic enzyme pyruvate kinase was significantly up-regulated (P<0·05) in the HL group. We also show that enhanced muscle protein synthesis and the transition from slow-to-fast fibres are altered by leucine. Together, these results indicate that leucine may alter energy metabolism and promote slow-to-fast transitions in the skeletal muscle of weanling piglets.

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Corresponding author

* Corresponding author: X. Yan, fax +86 27 87280408, email


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1. Wolfson, RL, Chantranupong, L, Saxton, RA, et al. (2016) Sestrin2 is a leucine sensor for the mTORC1 pathway. Science 351, 4348.
2. Sun, X & Zemel, MB (2009) Leucine modulation of mitochondrial mass and oxygen consumption in skeletal muscle cells and adipocytes. Nutr Metab (Lond) 6, 26.
3. D’Antona, G, Ragni, M, Cardile, A, et al. (2010) Branched-chain amino acid supplementation promotes survival and supports cardiac and skeletal muscle mitochondrial biogenesis in middle-aged mice. Cell Metab 12, 362372.
4. Nishitani, S, Matsumura, T, Fujitani, S, et al. (2002) Leucine promotes glucose uptake in skeletal muscles of rats. Biochem Biophys Res Commun 299, 693696.
5. Nair, KS, Woolf, PD, Welle, SL, et al. (1987) Leucine, glucose, and energy metabolism after 3 days of fasting in healthy human subjects. Am J Clin Nutr 46, 557562.
6. Chang, T & Goldberg, A (1978) Leucine inhibits oxidation of glucose and pyruvate in skeletal muscles during fasting. J Biol Chem 253, 36963701.
7. Paul, HS & Adibi, SA (1976) Assessment of effect of starvation, glucose, fatty acids and hormones on alpha-decarboxylation of leucine in skeletal muscle of rat. J Nutr 106, 10791088.
8. Li, H, Xu, M, Lee, J, et al. (2012) Leucine supplementation increases SIRT1 expression and prevents mitochondrial dysfunction and metabolic disorders in high-fat diet-induced obese mice. Am J Physiol Endocrinol Metab 303, E1234E1244.
9. Pette, D & Staron, RS (2000) Myosin isoforms, muscle fiber types, and transitions. Microsc Res Tech 50, 500509.
10. Schiaffino, S & Reggiani, C (1994) Myosin isoforms in mammalian skeletal muscle. J Appl Physiol 77, 493501.
11. Matsakas, A & Patel, K (2009) Skeletal muscle fibre plasticity in response to selected environmental and physiological stimuli. Histol Histopathol 24, 611629.
12. Davidsen, PK, Gallagher, IJ, Hartman, JW, et al. (2011) High responders to resistance exercise training demonstrate differential regulation of skeletal muscle microRNA expression. J Appl Physiol 110, 309317.
13. Ohlendieck, K (2010) Proteomics of skeletal muscle differentiation, neuromuscular disorders and fiber aging. Expert Rev Proteomics 7, 283296.
14. Reeds, PJ, Fjeld, CR & Jahoor, F (1994) Do the differences between the amino acid compositions of acute-phase and muscle proteins have a bearing on nitrogen loss in traumatic states? J Nutr 124, 906910.
15. Anthony, JC, Anthony, TG, Kimball, SR, et al. (2000) Orally administered leucine stimulates protein synthesis in skeletal muscle of postabsorptive rats in association with increased eIF4F formation. J Nutr 130, 139145.
16. Vary, TC (2007) Acute oral leucine administration stimulates protein synthesis during chronic sepsis through enhanced association of eukaryotic initiation factor 4G with eukaryotic initiation factor 4E in rats. J Nutr 137, 20742079.
17. Mobley, CB, Fox, CD, Thompson, RM, et al. (2016) Comparative effects of whey protein versus l-leucine on skeletal muscle protein synthesis and markers of ribosome biogenesis following resistance exercise. Amino Acids 48, 733750.
18. Escobar, J, Frank, JW, Suryawan, A, et al. (2005) Physiological rise in plasma leucine stimulates muscle protein synthesis in neonatal pigs by enhancing translation initiation factor activation. Am J Physiol Endocrinol Metab 288, E914E921.
19. Suryawan, A, Jeyapalan, AS, Orellana, RA, et al. (2008) Leucine stimulates protein synthesis in skeletal muscle of neonatal pigs by enhancing mTORC1 activation. Am J Physiol Endocrinol Metab 295, E868E875.
20. Hernandez-Garcia, AD, Columbus, DA, Manjarin, R, et al. (2016) Leucine supplementation stimulates protein synthesis and reduces degradation signal activation in muscle of newborn pigs during acute endotoxemia. Am J Physiol Endocrinol Metab 311, E791E801.
21. Koopman, R, Verdijk, L, Manders, RJ, et al. (2006) Co-ingestion of protein and leucine stimulates muscle protein synthesis rates to the same extent in young and elderly lean men. Am J Clin Nutr 84, 623632.
22. Bukhari, SS, Phillips, BE, Wilkinson, DJ, et al. (2015) Intake of low-dose leucine-rich essential amino acids stimulates muscle anabolism equivalently to bolus whey protein in older women at rest and after exercise. Am J Physiol Endocrinol Metab 308, E1056E1065.
23. Kramer, IF, Verdijk, LB, Hamer, HM, et al. (2016) Both basal and post-prandial muscle protein synthesis rates, following the ingestion of a leucine-enriched whey protein supplement, are not impaired in sarcopenic older males. Clin Nutr 16, 3126331268.
24. Stickland, N, Widdowson, EM & Goldspink, G (1975) Effects of severe energy and protein deficiencies on the fibres and nuclei in skeletal muscle of pigs. Br J Nutr 34, 421428.
25. Goldspink, G & Ward, P (1979) Changes in rodent muscle fibre types during post-natal growth, undernutrition and exercise. J Physiol 296, 453469.
26. Sieck, GC, Lewis, MI & Blanco, C (1989) Effects of undernutrition on diaphragm fiber size, SDH activity, and fatigue resistance. J Appl Physiol 66, 21962205.
27. Xia, Z, Cholewa, J, Zhao, Y, et al. (2016) Hypertrophy-promoting effects of leucine supplementation and moderate intensity aerobic exercise in pre-senescent mice. Nutrients 8, E246.
28. Okumura, N, Hashida-Okumura, A, Kita, K, et al. (2005) Proteomic analysis of slow- and fast-twitch skeletal muscles. Proteomics 5, 28962906.
29. Le Bihan, MC, Hou, Y, Harris, N, et al. (2006) Proteomic analysis of fast and slow muscles from normal and kyphoscoliotic mice using protein arrays, 2-DE and MS. Proteomics 6, 46464661.
30. Gelfi, C, Vigano, A, De Palma, S, et al. (2006) 2-D protein maps of rat gastrocnemius and soleus muscles: a tool for muscle plasticity assessment. Proteomics 6, 321340.
31. Gonnet, F, Bouazza, B, Millot, GA, et al. (2008) Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE). Proteomics 8, 264278.
32. Chan, XC, McDermott, JC & Siu, KW (2007) Identification of secreted proteins during skeletal muscle development. J Proteome Res 6, 698710.
33. Guelfi, KJ, Casey, TM, Giles, JJ, et al. (2006) A proteomic analysis of the acute effects of high-intensity exercise on skeletal muscle proteins in fasted rats. Clin Exp Pharmacol Physiol 33, 952957.
34. Almeida, AM, van Harten, S, Campos, A, et al. (2010) The effect of weight loss on protein profiles of gastrocnemius muscle in rabbits: a study using 1D electrophoresis and peptide mass fingerprinting. J Anim Physiol Anim Nutr (Berl) 94, 174185.
35. Hwang, H, Bowen, BP, Lefort, N, et al. (2010) Proteomics analysis of human skeletal muscle reveals novel abnormalities in obesity and type 2 diabetes. Diabetes 59, 3342.
36. Gelfi, C, Vigano, A, Ripamonti, M, et al. (2006) The human muscle proteome in aging. J Proteome Res 5, 13441353.
37. Gannon, J, Staunton, L, O’Connell, K, et al. (2008) Phosphoproteomic analysis of aged skeletal muscle. Int J Mol Med 22, 3342.
38. Wang, X, Ou, D, Yin, J, et al. (2009) Proteomic analysis reveals altered expression of proteins related to glutathione metabolism and apoptosis in the small intestine of zinc oxide-supplemented piglets. Amino Acids 37, 209218.
39. Goichon, A, Chan, P, Lecleire, S, et al. (2013) An enteral leucine supply modulates human duodenal mucosal proteome and decreases the expression of enzymes involved in fatty acid beta-oxidation. J Proteomics 78, 535544.
40. Oyedotun, KS & Lemire, BD (2004) The quaternary structure of the Saccharomyces cerevisiae succinate dehydrogenase homology modeling, cofactor docking, and molecular dynamics simulation studies. J Biol Chem 279, 94249431.
41. Hardie, DG, Ross, FA & Hawley, SA (2012) AMPK: a nutrient and energy sensor that maintains energy homeostasis. Nat Rev Mol Cell Biol 13, 251262.
42. Deacon, K & Blank, JL (1999) MEK kinase 3 directly activates MKK6 and MKK7, specific activators of the p38 and c-Jun NH2-terminal kinases. J Biol Chem 274, 1660416610.
43. Enslen, H, Raingeaud, J & Davis, RJ (1998) Selective activation of p38 mitogen-activated protein (MAP) kinase isoforms by the MAP kinase kinases MKK3 and MKK6. J Biol Chem 273, 17411748.
44. Hayes, JD & Pulford, DJ (1995) The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol 30, 445600.
45. McPherron, AC, Lawler, AM & Lee, S-J (1997) Regulation of skeletal muscle mass in mice by a new TGF-p superfamily member. Nature 387, 8390.
46. Newgard, CB (2012) Interplay between lipids and branched-chain amino acids in development of insulin resistance. Cell Metab 15, 606614.
47. Gerhart-Hines, Z, Rodgers, JT, Bare, O, et al. (2007) Metabolic control of muscle mitochondrial function and fatty acid oxidation through SIRT1/PGC‐1α . EMBO J 26, 19131923.
48. Fluck, M & Hoppeler, H (2003) Molecular basis of skeletal muscle plasticity – from gene to form and function. Rev Physiol Biochem Pharmacol 146, 159216.
49. Spangenburg, EE & Booth, FW (2003) Molecular regulation of individual skeletal muscle fibre types. Acta Physiol Scand 178, 413424.
50. Lombardi, A, Silvestri, E, Cioffi, F, et al. (2009) Defining the transcriptomic and proteomic profiles of rat ageing skeletal muscle by the use of a cDNA array, 2D- and Blue native-PAGE approach. J Proteomics 72, 708721.
51. Ohnuki, Y, Umeki, D, Cai, W, et al. (2013) Role of masseter muscle β 2-adrenergic signaling in regulation of muscle activity, myosin heavy chain transition, and hypertrophy. J Pharmacol Sci 123, 3646.
52. Muroya, S, Nakajima, I & Chikuni, K (2003) Amino acid sequences of multiple fast and slow troponin T isoforms expressed in adult bovine skeletal muscles. J Anim Sci 81, 11851192.
53. Perry, SV (2001) Vertebrate tropomyosin: distribution, properties and function. J Muscle Res Cell Motil 22, 549.
54. Burke, RE, Levine, DN & Zajac, FE 3rd (1971) Mammalian motor units: physiological-histochemical correlation in three types in cat gastrocnemius. Science 174, 709712.
55. Hara, K, Yonezawa, K, Weng, Q-P, et al. (1998) Amino acid sufficiency and mTOR regulate p70 S6 kinase and eIF-4E BP1 through a common effector mechanism. J Biol Chem 273, 1448414494.
56. Patti, M-E, Brambilla, E, Luzi, L, et al. (1998) Bidirectional modulation of insulin action by amino acids. J Clin Invest 101, 15191529.
57. Anthony, JC, Anthony, TG, Kimball, SR, et al. (2001) Signaling pathways involved in translational control of protein synthesis in skeletal muscle by leucine. J Nutr 131, 856S860S.
58. Elliott, B, Renshaw, D, Getting, S, et al. (2012) The central role of myostatin in skeletal muscle and whole body homeostasis. Acta Physiol (Oxf) 205, 324340.
59. Mendias, CL, Kayupov, E, Bradley, JR, et al. (2011) Decreased specific force and power production of muscle fibers from myostatin-deficient mice are associated with a suppression of protein degradation. J Appl Physiol (1985) 111, 185191.
60. Lokireddy, S, McFarlane, C, Ge, X, et al. (2011) Myostatin induces degradation of sarcomeric proteins through a Smad3 signaling mechanism during skeletal muscle wasting. Mol Endocrinol 25, 19361949.
61. Liu, X, Pan, S, Li, X, et al. (2015) Maternal low-protein diet affects myostatin signaling and protein synthesis in skeletal muscle of offspring piglets at weaning stage. Eur J Nutr 54, 971979.
62. Siriett, V, Platt, L, Salerno, MS, et al. (2006) Prolonged absence of myostatin reduces sarcopenia. J Cell Physiol 209, 866873.
63. Jackson, MF, Luong, D, Vang, DD, et al. (2012) The aging myostatin null phenotype: reduced adiposity, cardiac hypertrophy, enhanced cardiac stress response, and sexual dimorphism. J Endocrinol 213, 263275.
64. Lee, SJ & McPherron, AC (2001) Regulation of myostatin activity and muscle growth. Proc Natl Acad Sci U S A 98, 93069311.
65. Rebbapragada, A, Benchabane, H, Wrana, JL, et al. (2003) Myostatin signals through a transforming growth factor beta-like signaling pathway to block adipogenesis. Mol Cell Biol 23, 72307242.
66. Biesemann, N, Mendler, L, Wietelmann, A, et al. (2014) Myostatin regulates energy homeostasis in the heart and prevents heart failure. Circ Res 115, 296310.
67. Zhang, C, McFarlane, C, Lokireddy, S, et al. (2011) Myostatin-deficient mice exhibit reduced insulin resistance through activating the AMP-activated protein kinase signalling pathway. Diabetologia 54, 14911501.
68. Das, AK, Yang, QY, Fu, X, et al. (2012) AMP-activated protein kinase stimulates myostatin expression in C2C12 cells. Biochem Biophys Res Commun 427, 3640.
69. Philip, B, Lu, Z & Gao, Y (2005) Regulation of GDF-8 signaling by the p38 MAPK. Cell Signal 17, 365375.
70. Huang, Z, Chen, D, Zhang, K, et al. (2007) Regulation of myostatin signaling by c-Jun N-terminal kinase in C2C12 cells. Cell Signal 19, 22862295.
71. Morissette, MR, Cook, SA, Foo, S, et al. (2006) Myostatin regulates cardiomyocyte growth through modulation of Akt signaling. Circ Res 99, 1524.
72. Lin, JD, Handschin, C & Spiegelman, BM (2005) Metabolic control through the PGC-1 family of transcription coactivators. Cell Metab 1, 361370.
73. Finck, BN & Kelly, DP (2006) PGC-1 coactivators: inducible regulators of energy metabolism in health and disease. J Clin Invest 116, 615622.
74. Lin, J, Wu, H, Tarr, PT, et al. (2002) Transcriptional co-activator PGC-1 alpha drives the formation of slow-twitch muscle fibres. Nature 418, 797801.
75. Arany, Z, Lebrasseur, N, Morris, C, et al. (2007) The transcriptional coactivator PGC-1 beta drives the formation of oxidative type IIX fibers in skeletal muscle. Cell Metab 5, 3546.
76. Arany, Z (2008) PGC-1 coactivators and skeletal muscle adaptations in health and disease. Curr Opin Genet Dev 18, 426434.
77. Zong, HH, Ren, JM, Young, LH, et al. (2002) AMP kinase is required for mitochondrial biogenesis in skeletal muscle in response to chronic energy deprivation. Proc Natl Acad Sci U S A 99, 1598315987.
78. Roeckl, KS, Hirshman, MF, Brandauer, J, et al. (2007) Skeletal muscle adaptation to exercise training: AMP-activated protein kinase mediates muscle fiber type shift. Diabetes 56, 20622069.
79. Akimoto, T, Pohnert, SC, Li, P, et al. (2005) Exercise stimulates Pgc-1alpha transcription in skeletal muscle through activation of the p38 MAPK pathway. J Biol Chem 280, 1958719593.
80. Fan, M, Rhee, J, St-Pierre, J, et al. (2004) Suppression of mitochondrial respiration through recruitment of p160 myb binding protein to PGC-1 alpha: modulation by p38 MAPK. Genes Dev 18, 278289.
81. Puigserver, P, Rhee, J, Lin, J, et al. (2001) Cytokine stimulation of energy expenditure through p38 MAP kinase activation of PPARgamma coactivator-1. Mol Cell 8, 971982.
82. McKinsey, TA, Zhang, CL & Olson, EN (2002) MEF2: a calcium-dependent regulator of cell division, differentiation and death. Trends Biochem Sci 27, 4047.
83. Esser, K, Nelson, T, Lupa-Kimball, V, et al. (1999) The CACC box and myocyte enhancer factor-2 sites within the myosin light chain 2 slow promoter cooperate in regulating nerve-specific transcription in skeletal muscle. J Biol Chem 274, 1209512102.
84. Chin, ER, Olson, EN, Richardson, JA, et al. (1998) A calcineurin-dependent transcriptional pathway controls skeletal muscle fiber type. Genes Dev 12, 24992509.


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Dietary leucine supplementation alters energy metabolism and induces slow-to-fast transitions in longissimus dorsi muscle of weanling piglets

  • Qiwen Fan (a1) (a2) (a3), Baisheng Long (a1) (a2) (a3), Guokai Yan (a1) (a2) (a3), Zhichang Wang (a1) (a2) (a3), Min Shi (a1) (a2) (a3), Xiaoyu Bao (a1) (a2) (a3), Jun Hu (a1) (a2) (a3), Xiuzhi Li (a1) (a2) (a3), Changqing Chen (a1) (a2) (a3), Zilong Zheng (a1) (a2) (a3) and Xianghua Yan (a1) (a2) (a3)...


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