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http://dx.doi.org/10.5187/jast.2022.e45

Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress  

Jeong-Woong, Park (Department of Animal Science and Biotechnology, Kyungpook National University)
Kyoung Hwan, Kim (Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University)
Sujung, Kim (The Animal Molecular Genetics and Breeding Center, Jeonbuk National University)
Jae-rung, So (Department of Animal Science, Jeonbuk National University)
Byung-Wook, Cho (Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University)
Ki-Duk, Song (The Animal Molecular Genetics and Breeding Center, Jeonbuk National University)
Publication Information
Journal of Animal Science and Technology / v.64, no.4, 2022 , pp. 800-811 More about this Journal
Abstract
The integration of metabolomics and transcriptomics may elucidate the correlation between the genotypic and phenotypic patterns in organisms. In equine physiology, various metabolite levels vary during exercise, which may be correlated with a modified gene expression pattern of related genes. Integrated metabolomic and transcriptomic studies in horses have not been conducted to date. The objective of this study was to detect the effect of moderate exercise on the metabolomic and transcriptomic levels in horses. In this study, using nuclear magnetic resonance (NMR) spectroscopy, we analyzed the concentrations of metabolites in muscle and plasma; we also determined the gene expression patterns of branched chain (alpha) keto acid dehydrogenase kinase complex (BCKDK), which encodes the key regulatory enzymes in branched-chain amino acid (BCAA) catabolism, in two breeds of horses, Thoroughbred and Jeju, at different time intervals. The concentrations of metabolites in muscle and plasma were measured by 1H NMR (nuclear magnetic resonance) spectroscopy, and the relative metabolite levels before and after exercise in the two samples were compared. Subsequently, multivariate data analysis based on the metabolic profiles was performed using orthogonal partial least square discriminant analysis (OPLS-DA), and variable important plots and t-test were used for basic statistical analysis. The stress-induced expression patterns of BCKDK genes in horse muscle-derived cells were examined using quantitative reverse transcription polymerase chain reaction (qPCR) to gain insight into the role of transcript in response to exercise stress. In this study, we found higher concentrations of aspartate, leucine, isoleucine, and lysine in the skeletal muscle of Jeju horses than in Thoroughbred horses. In plasma, compared with Jeju horses, Thoroughbred horses had higher levels of alanine and methionine before exercise; whereas post-exercise, lysine levels were increased. Gene expression analysis revealed a decreased expression level of BCKDK in the post-exercise period in Thoroughbred horses.
Keywords
Metabolite; mRNA expression; Nuclear magnetic resonance (NMR) spectroscopy; Branched chain (alpha) keto acid dehydrogenase kinase complex (BCKDK) gene; Equine myoblast;
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1 Munoz A, Riber C, Trigo P, Castejon-Riber C, Castejon FM. Dehydration, electrolyte imbalances and renin-angiotensin-aldosterone-vasopressin axis in successful and unsuccessful endurance horses. Equine Vet J. 2010;42:83-90. https://doi.org/10.1111/j.2042-3306.2010.00211.x   DOI
2 Treiber KH, Hess TM, Kronfeld DS, Boston RC, Geor RJ, Friere M, et al. Glucose dynamics during exercise: dietary energy sources affect minimal model parameters in trained Arabian geldings during endurance exercise. Equine Vet J. 2006;38:631-6. https://doi.org/10.1111/j.2042-3306.2006.tb05617.x   DOI
3 Coenen M. Exercise and stress: impact on adaptive processes involving water and electrolytes. Livest Prod Sci. 2005;92:131-45. https://doi.org/10.1016/j.livprodsci.2004.11.018   DOI
4 Hargreaves M. Skeletal muscle metabolism during exercise in humans. Clin Exp Pharmacol Physiol. 2000;27:225-8. https://doi.org/10.1046/j.1440-1681.2000.03225.x   DOI
5 Beckonert O, Keun HC, Ebbels TMD, Bundy J, Holmes E, Lindon JC, et al. Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts. Nat Protoc. 2007;2:2692-703. https://doi.org/10.1038/nprot.2007.376   DOI
6 Keller MD, Pollitt CC, Marx UC. Nuclear magnetic resonance-based metabonomic study of early time point laminitis in an oligofructose-overload model. Equine Vet J. 2011;43:737-43. https://doi.org/10.1111/j.2042-3306.2010.00336.x   DOI
7 Ralston SL, Pappalardo L, Pelczer I, Spears PF. NMR-based metabonomic analyses of horse serum: detection of metabolic markers of disease. In: Recent recent advances in Animal Nutrition-Australia; 2011; Armidale, Australia. p. 197-205.
8 Apro W, Blomstrand E. Influence of supplementation with branched-chain amino acids in combination with resistance exercise on p70S6 kinase phosphorylation in resting and exercising human skeletal muscle. Acta Physiol. 2010;200:237-48. https://doi.org/10.1111/j.1748-1708.2010.02151.x   DOI
9 Anthony JC, Anthony TG, Kimball SR, Jefferson LS. Signaling pathways involved in translational control of protein synthesis in skeletal muscle by leucine. J Nutr. 2001;131:856S-60S. https://doi.org/10.1093/jn/131.3.856S   DOI
10 Du M, Shen QW, Zhu MJ, Ford SP. Leucine stimulates mammalian target of rapamycin signaling in C2C12 myoblasts in part through inhibition of adenosine monophosphateactivated protein kinase. J Anim Sci. 2007;85:919-27. https://doi.org/10.2527/jas.2006-342   DOI
11 Shimomura Y, Obayashi M, Murakami T, Harris RA. Regulation of branched-chain amino acid catabolism: nutritional and hormonal regulation of activity and expression of the branchedchain α-keto acid dehydrogenase kinase. Curr Opin Clin Nutr Metab Care. 2001;4:419-23. https://doi.org/10.1097/00075197-200109000-00013   DOI
12 Harris RA, Joshi M, Jeoung NH, Obayashi M. Overview of the molecular and biochemical basis of branched-chain amino acid catabolism. J Nutr. 2005;135:1527S-30S. https://doi.org/10.1093/jn/135.6.1527S   DOI
13 Suryawan A, Hawes JW, Harris RA, Shimomura Y, Jenkins AE, Hutson SM. A molecular model of human branched-chain amino acid metabolism. Am J Clin Nutr. 1998;68:72-81. https://doi.org/10.1093/ajcn/68.1.72   DOI
14 Popov KM, Zhao Y, Shimomura Y, Kuntz MJ, Harris RA. Branched-chainα-ketoacid dehydrogenase kinase. Molecular cloning, expression, and sequence similarity with histidine protein kinases. J Biol Chem. 1992;267:13127-30. https://doi.org/10.1016/S0021-9258(18)42179-5   DOI
15 Xu M, Nagasaki M, Obayashi M, Sato Y, Tamura T, Shimomura Y. Mechanism of activation of branched-chain α-keto acid dehydrogenase complex by exercise. Biochem Biophys Res Commun. 2001;287:752-6. https://doi.org/10.1006/bbrc.2001.5647   DOI
16 Le Moyec L, Robert C, Triba MN, Billat VL, Mata X, Schibler L, et al. Protein catabolism and high lipid metabolism associated with long-distance exercise are revealed by plasma NMR metabolomics in endurance horses. PLOS ONE. 2014;9:e90730. https://doi.org/10.1371/journal.pone.0090730   DOI
17 Nobukuni Y, Mitsubuchi H, Akaboshi I, Indo Y, Endo F, Yoshioka A, et al. Maple syrup urine disease. Complete defect of the E1 beta subunit of the branched chain alpha-ketoacid dehydrogenase complex due to a deletion of an 11-bp repeat sequence which encodes a mitochondrial targeting leader peptide in a family with the disease. J Clin Invest. 1991;87:1862-6. https://doi.org/10.1172/JCI115209   DOI
18 Garcia-Cazorla A, Oyarzabal A, Fort J, Robles C, Castejon E, Ruiz-Sala P, et al. Two novel mutations in the BCKDK (branched-chain keto-acid dehydrogenase kinase) gene are responsible for a neurobehavioral deficit in two pediatric unrelated patients. Hum Mutat. 2014;35:470-7. https://doi.org/10.1002/humu.22513   DOI
19 Trottier NL, Nielsen BD, Lang KJ, Ku PK, Schott HC. Equine endurance exercise alters serum branched-chain amino acid and alanine concentrations. Equine Vet J. 2002;34:168-72. https://doi.org/10.1111/j.2042-3306.2002.tb05412.x   DOI
20 Essen-Gustavsson B, Jensen-Waern M. Effect of an endurance race on muscle amino acids, pro- and macroglycogen and triglycerides. Equine Vet J. 2002;34:209-13. https://doi.org/10.1111/j.2042-3306.2002.tb05420.x   DOI
21 Park JW, Kim KH, Choi JK, Park TS, Song KD, Cho BW. Regulation of toll-like receptors expression in muscle cells by exercise-induced stress. Anim Biosci. 2021;34:1590-9. https://doi.org/10.5713/ab.20.0484   DOI
22 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25:402-8. https://doi.org/10.1006/meth.2001.1262   DOI
23 Sato T, Ito Y, Nagasawa T. Regulation of skeletal muscle protein degradation and synthesis by oral administration of lysine in rats. J Nutr Sci Vitaminol. 2013;59:412-9. https://doi.org/10.3177/jnsv.59.412   DOI
24 Jang HJ, Kim DM, Kim KB, Park JW, Choi JY, Oh JH, et al. Analysis of metabolomic patterns in thoroughbreds before and after exercise. Asian-Australas J Anim Sci. 2017;30:1633-42. https://doi.org/10.5713/ajas.17.0167   DOI
25 van den Hoven R, Bauer A, Hackl S, Zickl M, Spona J, Zentek J. Changes in intramuscular amino acid levels in submaximally exercised horses - a pilot study. J Anim Physiol Anim Nutr. 2010;94:455-64. https://doi.org/10.1111/j.1439-0396.2009.00929.x   DOI
26 Blomstrand E, Eliasson J, Karlsson HK, Kohnke R. Branched-chain amino acids activate key enzymes in protein synthesis after physical exercise. J Nutr. 2006;136:269S-73S. https://doi.org/10.1093/jn/136.1.269S   DOI
27 Assenza A, Bergero D, Tarantola M, Piccione G, Caola G. Blood serum branched chain amino acids and tryptophan modifications in horses competing in long-distance rides of different length. J Anim Physiol Anim Nutr. 2004;88:172-7. https://doi.org/10.1111/j.1439-0396.2004.00493.x   DOI
28 van den Hoven R, Bauer A, Hackl S, Zickl M, Spona J, Zentek J. A preliminary study on the changes in some potential markers of muscle-cell degradation in sub-maximally exercised horses supplemented with a protein and amino acid mixture. J Anim Physiol Anim Nutr. 2011;95:664-75. https://doi.org/10.1111/j.1439-0396.2010.01097.x   DOI
29 Barrey E, Evans SE, Evans DL, Curtis RA, Quinton R, Rose RJ. Locomotion evaluation for racing in Thoroughbreds. Equine Vet J. 2001;33:99-103. https://doi.org/10.1111/j.2042-3306.2001.tb05369.x   DOI
30 Obayashi M, Sato Y, Harris RA, Shimomura Y. Regulation of the activity of branched-chain 2-oxo acid dehydrogenase (BCODH) complex by binding BCODH kinase. FEBS Lett. 2001;491:50-4. https://doi.org/10.1016/S0014-5793(01)02149-4   DOI