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Changes in metabolites following extreme alterations in the muscle environment in horses after exercise may provide useful evidence. We compared the muscle metabolites before and after supramaximal exercise to fatigue in horses. Six well-trained horses ran until exhaustion in incremental exercise tests. Biopsy samples were obtained from the <jats:italic>gluteus medius<\/jats:italic> muscle before and immediately after exercise for capillary electrophoresis\u2013mass spectrometry analysis. In the incremental exercise test, the total running time and speed of the last step were 10.4\u2009\u00b1\u20091.3 (mean\u2009\u00b1\u2009standard deviation) min and 12.7\u2009\u00b1\u20090.5\u00a0m\/s, respectively. Of 73 metabolites, 18 and 11 were significantly increased and decreased after exercise, respectively. The heat map of the hierarchical cluster analysis of muscle metabolites showed that changes in metabolites were clearly distinguishable before and after exercise. Strenuous exercise increased many metabolites in the glycolytic pathway and the tricarboxylic acid cycle in skeletal muscle. Targeted metabolomic analysis of skeletal muscle may clarify the intramuscular environment caused by exercise and explain the response of working muscles to strenuous exercise that induces hypoxemia and hypercapnia in Thoroughbred horses.<\/jats:p>","DOI":"10.1038\/s41598-021-90834-y","type":"journal-article","created":{"date-parts":[[2021,5,27]],"date-time":"2021-05-27T10:16:36Z","timestamp":1622110596000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Metabolomic analysis of skeletal muscle before and after strenuous exercise to fatigue"],"prefix":"10.1038","volume":"11","author":[{"given":"Hajime","family":"Ohmura","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazutaka","family":"Mukai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuji","family":"Takahashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshiyuki","family":"Takahashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,5,27]]},"reference":[{"key":"90834_CR1","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1186\/1751-0147-55-59","volume":"55","author":"H Ohmura","year":"2013","unstructured":"Ohmura, H., Matsui, A., Hada, T. & Jones, J. 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