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This study investigated muscle metabolic responses to positive and negative pacing strategies in Thoroughbred horses. Eight Thoroughbred horses performed 2\u00a0min treadmill running using positive (1\u00a0min at 110% maximal O<jats:sub>2<\/jats:sub> uptake [V\u0307O<jats:sub>2<\/jats:sub>max], followed by 1\u00a0min at 90% V\u0307O<jats:sub>2<\/jats:sub>max) and negative (1\u00a0min at 90% V\u0307O<jats:sub>2<\/jats:sub>max, followed by 1\u00a0min at 110% V\u0307O<jats:sub>2<\/jats:sub>max) pacing strategies. The arterial-mixed venous O<jats:sub>2<\/jats:sub> difference did not significantly differ between the two strategies. Plasma lactate levels increased toward 2\u00a0min, with significantly higher concentrations during positive pacing than during negative pacing. Muscle glycogen level was significantly lower at 1 and 2\u00a0min of positive pacing than those of negative pacing. Metabolomic analysis showed that the sum of glycolytic intermediates increased during the first half of positive pacing and the second half of negative pacing. Regardless of pacing strategy, the sum of tricarboxylic acid cycle metabolites increased during the first half but remained unchanged thereafter. Our data suggest that positive pacing strategy is likely to activate glycolytic metabolism to a greater extent compared to negative pacing, even though the total workload is identical.<\/jats:p>","DOI":"10.1038\/s41598-024-69339-x","type":"journal-article","created":{"date-parts":[[2024,8,7]],"date-time":"2024-08-07T15:02:24Z","timestamp":1723042944000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Effects of pacing strategy on metabolic responses to 2-min intense exercise in Thoroughbred horses"],"prefix":"10.1038","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8643-6849","authenticated-orcid":false,"given":"Kenya","family":"Takahashi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1992-6634","authenticated-orcid":false,"given":"Kazutaka","family":"Mukai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9165-6835","authenticated-orcid":false,"given":"Yusaku","family":"Ebisuda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fumi","family":"Sugiyama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshinobu","family":"Yoshida","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1428-5911","authenticated-orcid":false,"given":"Hideo","family":"Hatta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6932-2735","authenticated-orcid":false,"given":"Yu","family":"Kitaoka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,8,7]]},"reference":[{"key":"69339_CR1","doi-asserted-by":"publisher","first-page":"239","DOI":"10.2165\/00007256-200838030-00004","volume":"38","author":"CR Abbiss","year":"2008","unstructured":"Abbiss, C. 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