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Life (Basel, Switzerland)2021; 11(7); doi: 10.3390/life11070719

In Vitro Assays for the Assessment of Impaired Mitochondrial Bioenergetics in Equine Atypical Myopathy.

Abstract: Equine atypical myopathy is a seasonal intoxication of grazing equids. In Europe, this poisoning is associated with the ingestion of toxins contained in the seeds and seedlings of the sycamore maple (Acer pseudoplatanus). The toxins involved in atypical myopathy are known to inhibit ß-oxidation of fatty acids and induce a general decrease in mitochondrial respiration, as determined by high-resolution respirometry applied to muscle samples taken from cases of atypical myopathy. The severe impairment of mitochondrial bioenergetics induced by the toxins may explain the high rate of mortality observed: about 74% of horses with atypical myopathy die, most within the first two days of signs of poisoning. The mechanism of toxicity is not completely elucidated yet. To improve our understanding of the pathological process and to assess therapeutic candidates, we designed in vitro assays using equine skeletal myoblasts cultured from muscle biopsies and subjected to toxins involved in atypical myopathy. We established that equine primary myoblasts do respond to one of the toxins incriminated in the disease.
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Publication Date: 2021-07-20 PubMed ID: 34357091PubMed Central: PMC8307747DOI: 10.3390/life11070719Google Scholar: Lookup
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  • 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 examines the impact of toxins associated with Equine Atypical Myopathy (a type of poisoning) on the efficiency of mitochondria in horse muscle cells. The study designs tests using cultured horse muscle cells exposed to these toxins.

Research Overview

Equine Atypical Myopathy (EAM) is a seasonally occurring intoxication found in grazing equids, primarily horses, and is particularly common in Europe. It’s associated with ingestion of toxins found in seeds and seedlings of the sycamore maple. The toxins related to EAM are known to disrupt the ß-oxidation of fatty acids, which is critical for energy production, thereby decreasing the efficiency of mitochondria – the powerhouses of cells. Using high-resolution respirometry on samples gathered from EAM cases, a significant decrease in mitochondrial respiration is observed. A mortality rate of about 74% is seen in horses with EAM, with most of them dying within the first two days of showing poisoning signs. However, the specific toxicity mechanism is not yet fully understood.

Methodology

  • The researchers aimed to enhance understanding of the pathological process and evaluate potential therapeutic measures.
  • The study designed in vitro assays (experiments conducted in controlled artificial environments), using equine skeletal myoblasts (precursor muscle cells) cultured from muscle biopsies and subjected to toxins associated with EAM. These tests were designed to mimic the in vivo (within the body) environment of EAM intoxication.

Findings

  • The researchers established that equine primary myoblasts do respond to one of the toxins implicated in EAM. This suggests the toxin has a direct impact on the precursor muscle cells, which might explain the high rate of muscle degradation observed in afflicted horses.
  • The research’s findings and in vitro experiments lay the groundwork for more detailed studies on horse muscle cell responses to various toxins, potentially enabling better understanding and treatment for EAM.

Cite This Article

APA
Kruse CJ, Stern D, Mouithys-Mickalad A, Niesten A, Art T, Lemieux H, Votion DM. (2021). In Vitro Assays for the Assessment of Impaired Mitochondrial Bioenergetics in Equine Atypical Myopathy. Life (Basel), 11(7). https://doi.org/10.3390/life11070719

Publication

Life (Basel, Switzerland)
ISSN: 2075-1729
NlmUniqueID: 101580444
Country: Switzerland
Language: English
Volume: 11
Issue: 7

Researcher Affiliations

Kruse, Caroline-J
  • Department of Functional Sciences, Faculty of Veterinary Medicine, Physiology and Sport Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Stern, David
  • Equine Pole, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medicine, University of Liège, 4000 Liège, Belgium.
Mouithys-Mickalad, Ange
  • Center for Oxygen, Research & Development (CORD), Center for Interdisciplinary Research on Medicines (CIRM), Institute of Chemistry, B6a, University of Liège, Allée du Six Août, 11, 4000 Liège, Belgium.
Niesten, Ariane
  • Center for Oxygen, Research & Development (CORD), Fundamental and Applied Research for Animals & Health (FARAH), Institute of Chemistry, B6a, University of Liège, Allée du Six Août, 11, 4000 Liège, Belgium.
Art, Tatiana
  • Department of Functional Sciences, Faculty of Veterinary Medicine, Physiology and Sport Medicine, Fundamental and Applied Research for Animals & Health (FARAH), University of Liège, 4000 Liège, Belgium.
Lemieux, Hélène
  • Faculty Saint-Jean and Department of Medicine, University of Alberta, 8406-91 Street, Edmonton, AB T6C 4G9, Canada.
Votion, Dominique-M
  • Equine Pole, Fundamental and Applied Research for Animals & Health (FARAH), Faculty of Veterinary Medicine, University of Liège, 4000 Liège, Belgium.

Grant Funding

  • 40001860 / Fonds De La Recherche Scientifique - FNRS
  • R.CFRA.2991-J-F / Fonds spéciaux à la recherche, Université de Liège

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 37 references
  1. Votion D.-M., van Galen G., Sweetman L., Boemer F., de Tullio P., Dopagne C., Lefère L., Mouithys-Mickalad A., Patarin F., Rouxhet S.. Identification of methylenecyclopropyl acetic acid in serum of European horses with atypical myopathy.. Equine Vet. J. 2013;46:146–149.
    doi: 10.1111/evj.12117pubmed: 23773055google scholar: lookup
  2. Valberg S.J., Sponseller B.T., Hegeman A.D., Earing J., Bender J.B., Martinson K.L., Patterson S.E., Sweetman L.. Seasonal pasture myopathy/atypical myopathy in North America associated with ingestion of hypoglycin A within seeds of the box elder tree.. Equine Vet. J. 2013;45:419–426.
    doi: 10.1111/j.2042-3306.2012.00684.xpubmed: 23167695google scholar: lookup
  3. Unger L., Nicholson A., Jewitt E.M., Gerber V., Hegeman A., Sweetman L., Valberg S.. Hypoglycin A concentrations in seeds of acer pseudoplatanus trees growing on atypical myopathy-affected and control pastures.. J. Vet. Intern. Med. 2014;28:1289–1290.
    doi: 10.1111/jvim.12367pmc: PMC4857957pubmed: 24863395google scholar: lookup
  4. Westermann C.M., van Leeuwen R., van Raamsdonk L.W.D., Mol H.G.J.. Hypoglycin A concentrations in maple tree species in the netherlands and the occurrence of atypical myopathy in horses.. J. Vet. Intern. Med. 2016;30:880–884.
    doi: 10.1111/jvim.13927pmc: PMC4913566pubmed: 26995161google scholar: lookup
  5. Baise E., Habyarimana J.A., Amory H., Boemer F., Douny C., Gustin P., Marcillaud-Pitel C., Patarin F., Weber M., Votion D.M.. Samaras and seedlings of Acer pseudoplatanus are potential sources of hypoglycin A intoxication in atypical myopathy without necessarily inducing clinical signs.. Equine Vet. J. 2016;48:414–417.
    doi: 10.1111/evj.12499pubmed: 26278545google scholar: lookup
  6. Fowden L., Pratt H.M.. Cyclopropyl amino acids of the genus Acer: Distribution and biosynthesis.. Phytochemistry 1973;12:1677–1681.
    doi: 10.1016/0031-9422(73)80387-5google scholar: lookup
  7. Bochnia M., Sander J., Ziegler J., Terhardt M., Sander S., Janzen N., Cavalleri J.V., Zuraw A., Wensch-Dorendorf M., Zeyner A.. Detection of MCPG metabolites in horses with atypical myopathy.. PLoS ONE 2019;14:e0211698.
    doi: 10.1371/journal.pone.0211698pmc: PMC6363182pubmed: 30721263google scholar: lookup
  8. Von Holt C.. Methylenecyclopropaneacetic acid, a metabolite of hypoglycin.. Biochim. Biophys. Acta 1966;125:1–10.
    doi: 10.1016/0005-2760(66)90138-Xpubmed: 5968592google scholar: lookup
  9. Melde K., Buettner H., Boschert W., Wolf H.P., Ghisla S.. Mechanism of hypoglycaemic action of methylenecyclopropylglycine.. Biochem. J. 1989;259:921–924.
    doi: 10.1042/bj2590921pmc: PMC1138607pubmed: 2730593google scholar: lookup
  10. Melde K., Jackson S., Bartlett K., Stanley H., Sherrattt A., Ghisla S.. Metabolic consequences of methylenecyclopropylglycine poisoning in rats.. Biochem. J. 1991;274:395–400.
    doi: 10.1042/bj2740395pmc: PMC1150150pubmed: 2006907google scholar: lookup
  11. Von Holt C., Chang J., Von Holt M., Böhm H.. Metabolism and metabolic effects of hypoglycin.. Biochim. Biophys. Acta 1964;90:611–613.
    doi: 10.1016/0304-4165(64)90242-9pubmed: 14237871google scholar: lookup
  12. Westermann C.M., Dorland D., van Diggelen O.P., Schoonderwoerd K., Bierau J., Waterham H.R., van der Kolk J.H.. Decreased oxidative phosphorylation and PGAM deficiency in horses suffering from atypical myopathy associated with acquired MADD.. Mol. Gen. Metab. 2011;104:273–278.
    doi: 10.1016/j.ymgme.2011.07.022pubmed: 21843962google scholar: lookup
  13. Lemieux H., Boemer F., van Galen G., Serteyn D., Amory H., Baise E., Cassart D., van Loon G., Marcillaud-Pitel C., Votion D.M.. Mitochondrial function is altered in horse atypical myopathy.. Mitochondrion 2016;30:35–41.
    doi: 10.1016/j.mito.2016.06.005pubmed: 27374763google scholar: lookup
  14. Westermann C.M., Dorland L., Votion D.M., de Sain-van der Velden M.G., Wijnberg I.D., Wanders R.J., Spliet W.G., Testerink N., Berger R., Ruiter J.P.. Acquired multiple Acyl-CoA dehydrogenase deficiency in 10 horses with atypical myopathy.. Neuromuscul. Disord. 2008;18:355–364.
    doi: 10.1016/j.nmd.2008.02.007pubmed: 18406615google scholar: lookup
  15. Bochnia M., Ziegler J., Sander J., Uhlig A., Schaefer S., Vollstedt S., Glatter M., Abel S., Recknagel S., Schusser G.F.. Hypoglycin A content in blood and urine discriminates horses with atypical myopathy from clinically normal horses grazing on the same pasture.. PLoS ONE 2015;10:e0136785.
    doi: 10.1371/journal.pone.0136785pmc: PMC4574941pubmed: 26378918google scholar: lookup
  16. Karlíková R., Široká J., Jahn P., Friedecký D., Gardlo A., Janečková H., Hrdinová F., Drábková Z., Adam T.. Equine atypical myopathy: A metabolic study.. Vet. J. 2016;216:125–132.
    doi: 10.1016/j.tvjl.2016.07.015pubmed: 27687939google scholar: lookup
  17. Boemer F., Detilleux J., Cello C., Amory H., Marcillaud-Pitel C., Richard E., van Galen G., van Loon G., Lefère L., Votion D.M.. Acylcarnitines profile best predicts survival in horses with atypical myopathy.. PLoS ONE 2017;12:e0182761.
    doi: 10.1371/journal.pone.0182761pmc: PMC5573150pubmed: 28846683google scholar: lookup
  18. van Galen G., Marcillaud Pitel C., Saegerman C., Patarin F., Amory H., Baily J.D., Cassart D., Gerber V., Hahn C., Harris P.. European outbreaks of atypical myopathy in grazing equids (2006–2009): Spatiotemporal distribution, history and clinical features.. Equine Vet. J. 2012;44:614–620.
    doi: 10.1111/j.2042-3306.2012.00556.xpubmed: 22448904google scholar: lookup
  19. Cassart D., Baise E., Cherel Y., Delguste C., Antoine N., Votion D., Amory H., Rollin F., Linden A., Coignoul F.. Morphological alterations in oxidative muscles and mitochondrial structure associated with equine atypical myopathy.. Equine Vet. J. 2007;39:26–32.
    doi: 10.2746/042516407X157765pubmed: 17228591google scholar: lookup
  20. Palencia P., Rivero J.L.L.. Atypical myopathy in two grazing equids in northern Spain.. Vet. Rec. 2007;161:346–348.
    doi: 10.1136/vr.161.10.346pubmed: 17827475google scholar: lookup
  21. Votion D.-M., Gnaiger E., Lemieux H., Mouithys-Mickalad A., Serteyn S.. Physical fitness and mitochondrial respiratory capacity in horse skeletal muscle.. PLoS ONE 2012;7:e34890.
    doi: 10.1371/annotation/e24ced68-9c2d-4303-9553-0661decb9a51pmc: PMC3329552pubmed: 22529950google scholar: lookup
  22. Votion D.M., François A.C., Kruse C., Renaud B., Farinelle A., Bouquieaux M.C., Marcillaud-Pitel C., Gustin P.. Answers to the frequently asked questions regarding horse feeding and management practices to reduce the risk of atypical myopathy.. Animals 2020;24:365.
    doi: 10.3390/ani10020365pmc: PMC7071031pubmed: 32102384google scholar: lookup
  23. van Galen G., Saegerman C., Marcillaud Pitel C., Patarin F., Amory H., Baily J.D., Cassart D., Gerber V., Hahn C., Harris P.. European outbreaks of atypical myopathy in grazing horses (2006–2009): Determination of indicators for risk and prognostic factors.. Equine Vet. J. 2012;44:621–625.
    doi: 10.1111/j.2042-3306.2012.00555.xpubmed: 22413891google scholar: lookup
  24. Chen K.K., Anderson R.C., McCowen M.C., Harris P.N.. Pharmacologic action of hypoglycin A and B.. J. Pharmacol. Exp. Ther. 1957;121:272–285.
    pubmed: 13481850
  25. Brooks S.E., Audretsch J.J.. Studies on hypoglycin toxicity in rats. I. Changes in hepatic ultrastructure.. Am. J. Pathol. 1970;59:161–180.
    pmc: PMC2032857pubmed: 5441716
  26. Ceusters J.D., Mouithys-Mickalad A.A., de la Rebière de Pouyade G., Franck T.J., Votion D.M., Deby-Dupont G.P., Serteyn D.A.. Assessment of reactive oxygen species production in cultured equine skeletal myoblasts in response to conditions of anoxia followed by reoxygenation with or without exposure to peroxidases.. Am. J. Vet. Res. 2012;73:426–434.
    doi: 10.2460/ajvr.73.3.426pubmed: 22369537google scholar: lookup
  27. Pesta D., Gnaiger E.. High-Resolution Respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle.. Methods Mol. Biol. 2012;810:25–58.
    doi: 10.1007/978-1-61779-382-0_3pubmed: 22057559google scholar: lookup
  28. Gnaiger E.. Mitochondrial Pathways and Respiratory Control. An Introduction to OXPHOS Analysis.. 5th ed. Bioenergetics Communications; Axams, Austria: 2020.
    doi: 10.26124/bec:2020-0002google scholar: lookup
  29. Lemieux H., Subarsky P., Doblander C., Wurn M., Troppmair J., Gnaiger E.. Impairment of mitochondrial respiratory function as an early biomarker of apoptosis induced by growth factor removal.. BioRxiv 2017.
    doi: 10.1101/151480google scholar: lookup
  30. Remels A.H.V., Langen R.C.J., Schrauwen P., Schaart G., Schols A.M.W.J., Gosker H.R.. Regulation of mitochondrial biogenesis during myogenesis.. Mol. Cell. Endocrinol. 2010;315:113–120.
    doi: 10.1016/j.mce.2009.09.029pubmed: 19804813google scholar: lookup
  31. Bavli D., Prill S., Ezra E., Levy G., Cohen M., Vinken M., Vanfleteren J., Jaeger M., Nahmias Y.. Real-time monitoring of metabolic function in liver-onchip microdevices tracks the dynamics of Mitochondrial dysfunction.. Proc. Natl. Acad. Sci. USA 2016;113:E2231–E2240.
    doi: 10.1073/pnas.1522556113pmc: PMC4843487pubmed: 27044092google scholar: lookup
  32. Schrauwen P., Schrauwen-Hinderling V., Hoeks J., Hesselink M.K.C.. Mitochondrial dysfunction and lipotoxicity.. BBA Mol. Cell. Biol. Lipids 2010;1801:266–271.
    doi: 10.1016/j.bbalip.2009.09.011pubmed: 19782153google scholar: lookup
  33. Cui H., Kong Y., Zhang H.. Oxidative Stress, Mitochondrial Dysfunction, and Aging.. J. Signal Trans. 2012;2012:1–13.
    doi: 10.1155/2012/646354pmc: PMC3184498pubmed: 21977319google scholar: lookup
  34. Farah B.L., Sinha R.A., Wu Y., Singh B.K., Lim A., Hirayama M., Landau D.J., Bay B.H., Koeberl D., Yen P.M.. Hepatic mitochondrial dysfunction is a feature of Glycogen Storage Disease Type Ia (GSDIa). Sci. Rep. 2017;7:1–12.
    doi: 10.1038/srep44408pmc: PMC5357851pubmed: 28317891google scholar: lookup
  35. Moehle E.A., Shen K., Dillin A.. Mitochondrial proteostasis in the context of cellular and organismal health and aging.. J. Biol. Chem. 2019;14:5396–5407.
    doi: 10.1074/jbc.TM117.000893pmc: PMC6462515pubmed: 29622680google scholar: lookup
  36. Baydoun A.R.. Cell culture techniques.. In: Baydoun A.R., Wilson K., editors. Principles and Techniques of Biochemistry and Molecular Biology. Cambridge University Press; Cambridge, UK: 2005. pp. 71–102.
    doi: 10.1017/CBO9780511813412.003google scholar: lookup
  37. Kaur G., Dufour J.M.. Cell lines.. Spermatogenesis 2012;2:1–5.
    doi: 10.4161/spmg.19885pmc: PMC3341241pubmed: 22553484google scholar: lookup

Citations

This article has been cited 3 times.
  1. François AC, Taminiau B, Renaud B, Gonza-Quito IE, Massey C, Hyde C, Piercy RJ, Douny C, Scippo ML, Daube G, Gustin P, Delcenserie V, Votion DM. In Vitro Investigation of Equine Gut Microbiota Alterations During Hypoglycin A Exposure. Animals (Basel) 2025 Nov 19;15(22).
    doi: 10.3390/ani15223343pubmed: 41302050google scholar: lookup
  2. Wouters CP, Klein B, Price N, Boemer F, Voz ML, Votion DM. A Zebrafish Embryo Model to Screen Potential Therapeutic Compounds in Sapindaceae Poisoning. Molecules 2024 Oct 19;29(20).
    doi: 10.3390/molecules29204954pubmed: 39459322google scholar: lookup
  3. Nesci S, Lenaz G. Impaired Mitochondrial Bioenergetics under Pathological Conditions. Life (Basel) 2022 Jan 29;12(2).
    doi: 10.3390/life12020205pubmed: 35207491google scholar: lookup
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