Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress.
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 (), 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 H 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 -test were used for basic statistical analysis. The stress-induced expression patterns of 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 in the post-exercise period in Thoroughbred horses.
© Copyright 2022 Korean Society of Animal Science and Technology.
Publication Date: 2022-07-31 PubMed ID: 35969708PubMed Central: PMC9353351DOI: 10.5187/jast.2022.e45Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research article investigates how moderate exercise impacts levels of metabolites (substances needed for metabolism) and gene expression in horses. By utilising nuclear magnetic resonance (NMR) spectroscopy to analyse metabolites in muscle and plasma, and evaluating gene expression of branched chain (alpha) keto acid dehydrogenase kinase complex, the study found differences in concentrations of certain metabolites and changes in gene expression after exercise between two horse breeds, Thoroughbred and Jeju.
Research Objectives and Methods
- The researchers sought to understand the effects of exercise on the metabolomic (study of the chemical processes involved in metabolism) and transcriptomic (study of the complete set of RNA molecules) levels in horses. This is the first study to combine these two areas of study in equine physiology.
- Nuclear magnetic resonance (NMR) spectroscopy was used to analyse concentrations of metabolites in muscle and plasma. This non-invasive technique provides detailed information about the structure and dynamics of molecules.
- Gene expression patterns of branched chain (alpha) keto acid dehydrogenase kinase complex were determined using quantitative reverse transcription polymerase chain reaction (qPCR). This gene complex is vital to the metabolism of branched-chain amino acids (BCAAs), which are essential nutrients.
Findings
- The study discovered differences in metabolite levels and gene expression between the two breeds of horses both before and after exercise.
- Jeju horses displayed higher concentrations of aspartate, leucine, isoleucine, and lysine in skeletal muscle compared to Thoroughbred horses.
- Thoroughbred horses had higher levels of alanine and methionine in plasma before exercise compared to Jeju horses. After exercise, lysine levels in plasma increased in Thoroughbreds.
- Exercise led to changes in the gene expression of branched chain (alpha) keto acid dehydrogenase kinase complex. Expression levels decreased post-exercise in Thoroughbred horses, suggesting a potential impact of exercise on the catabolism (breakdown) of BCAAs.
Implications
- The study suggests that moderate exercise might cause changes in metabolite levels and gene expression in horses. This could have implications for equine health and performance.
- Differences between the two horse breeds indicate a possible genetic divergence in metabolite concentration and gene expression, which could be explored in subsequent studies. Understanding these differences may help refine exercise and diet regimes for different breeds to optimise their health and performance.
Cite This Article
APA
Park JW, Kim KH, Kim S, So JR, Cho BW, Song KD.
(2022).
Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress.
J Anim Sci Technol, 64(4), 800-811.
https://doi.org/10.5187/jast.2022.e45 Publication
Researcher Affiliations
- Department of Animal Science and Biotechnology, Kyungpook National University, SangJu 37224, Korea.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
- The Animal Molecular Genetics and Breeding Center, Jeonbuk National University, Jeonju 54896, Korea.
- Department of Animal Science, Jeonbuk National University, Jeonju 54896, Korea.
- Department of Animal Science, College of Natural Resources and Life Sciences, Pusan National University, Miryang 50463, Korea.
- The Animal Molecular Genetics and Breeding Center, Jeonbuk National University, Jeonju 54896, Korea.
- Department of Agricultural Convergence Technology, Jeonbuk National University, Jeonju 54896, Korea.
Conflict of Interest Statement
No potential conflict of interest relevant to this article was reported.
References
This article includes 30 references
- Muñoz A, Riber C, Trigo P, Castejón-Riber C, Castejón FM. Dehydration, electrolyte imbalances and renin-angiotensin-aldosterone-vasopressin axis in successful and unsuccessful endurance horses.. Equine Vet J Suppl 2010 Nov;(38):83-90.
- Treiber KH, Hess TM, Kronfeld DS, Boston RC, Geor RJ, Friere M, Silva AM, Harris PA. Glucose dynamics during exercise: dietary energy sources affect minimal model parameters in trained Arabian geldings during endurance exercise.. Equine Vet J Suppl 2006 Aug;(36):631-6.
- Coenen M. Exercise and stress: impact on adaptive processes involving water and electrolytes.. Livest Prod Sci 2005;92:131–45.
- Hargreaves M. Skeletal muscle metabolism during exercise in humans.. Clin Exp Pharmacol Physiol 2000 Mar;27(3):225-8.
- Beckonert O, Keun HC, Ebbels TM, Bundy J, Holmes E, Lindon JC, Nicholson JK. Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts.. Nat Protoc 2007;2(11):2692-703.
- 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 Nov;43(6):737-43.
- Ralston SL, Pappalardo L, Pelczer I, Spears PF. Recent recent advances in Animal Nutrition-Australia. Armidale, Australia: 2011. NMR-based metabonomic analyses of horse serum: detection of metabolic markers of disease; pp. 197–205.
- Apró 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 (Oxf) 2010 Nov;200(3):237-48.
- 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 Mar;131(3):856S-860S.
- Du M, Shen QW, Zhu MJ, Ford SP. Leucine stimulates mammalian target of rapamycin signaling in C2C12 myoblasts in part through inhibition of adenosine monophosphate-activated protein kinase.. J Anim Sci 2007 Apr;85(4):919-27.
- 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 branched-chain alpha-keto acid dehydrogenase kinase.. Curr Opin Clin Nutr Metab Care 2001 Sep;4(5):419-23.
- Harris RA, Joshi M, Jeoung NH, Obayashi M. Overview of the molecular and biochemical basis of branched-chain amino acid catabolism.. J Nutr 2005 Jun;135(6 Suppl):1527S-30S.
- 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 Jul;68(1):72-81.
- Popov KM, Zhao Y, Shimomura Y, Kuntz MJ, Harris RA. Branched-chain alpha-ketoacid dehydrogenase kinase. Molecular cloning, expression, and sequence similarity with histidine protein kinases.. J Biol Chem 1992 Jul 5;267(19):13127-30.
- Xu M, Nagasaki M, Obayashi M, Sato Y, Tamura T, Shimomura Y. Mechanism of activation of branched-chain alpha-keto acid dehydrogenase complex by exercise.. Biochem Biophys Res Commun 2001 Sep 28;287(3):752-6.
- Nobukuni Y, Mitsubuchi H, Akaboshi I, Indo Y, Endo F, Yoshioka A, Matsuda I. 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 May;87(5):1862-6.
- García-Cazorla A, Oyarzabal A, Fort J, Robles C, Castejón E, Ruiz-Sala P, Bodoy S, Merinero B, Lopez-Sala A, Dopazo J, Nunes V, Ugarte M, Artuch R, Palacín M, Rodríguez-Pombo P, Alcaide P, Navarrete R, Sanz P, Font-Llitjós M, Vilaseca MA, Ormaizabal A, Pristoupilova A, Agulló SB. 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 Apr;35(4):470-7.
- 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 Suppl 2002 Sep;(34):168-72.
- Le Moyec L, Robert C, Triba MN, Billat VL, Mata X, Schibler L, Barrey E. Protein catabolism and high lipid metabolism associated with long-distance exercise are revealed by plasma NMR metabolomics in endurance horses.. PLoS One 2014;9(3):e90730.
- Essén-Gustavsson B, Jensen-Waern M. Effect of an endurance race on muscle amino acids, pro- and macroglycogen and triglycerides.. Equine Vet J Suppl 2002 Sep;(34):209-13.
- 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 Oct;34(10):1590-1599.
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.. Methods 2001 Dec;25(4):402-8.
- Jang HJ, Kim DM, Kim KB, Park JW, Choi JY, Oh JH, Song KD, Kim S, Cho BW. Analysis of metabolomic patterns in thoroughbreds before and after exercise.. Asian-Australas J Anim Sci 2017 Nov;30(11):1633-1642.
- 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 (Berl) 2010 Aug 1;94(4):455-64.
- Blomstrand E, Eliasson J, Karlsson HK, Köhnke R. Branched-chain amino acids activate key enzymes in protein synthesis after physical exercise.. J Nutr 2006 Jan;136(1 Suppl):269S-73S.
- 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 (Tokyo) 2013;59(5):412-9.
- 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 (Berl) 2004 Apr;88(3-4):172-7.
- 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 (Berl) 2011 Oct;95(5):664-75.
- Barrey E, Evans SE, Evans DL, Curtis RA, Quinton R, Rose RJ. Locomotion evaluation for racing in thoroughbreds.. Equine Vet J Suppl 2001 Apr;(33):99-103.
- 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 Feb 23;491(1-2):50-4.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
