{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T04:01:06Z","timestamp":1792814466268,"version":"4.5.0"},"reference-count":60,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,8,21]],"date-time":"2020-08-21T00:00:00Z","timestamp":1597968000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Spanish Ministry of Science, Innovation and Universities","award":["RTI2018-096736-A-I00 and RTI2018-093548-B-I00"],"award-info":[{"award-number":["RTI2018-096736-A-I00 and RTI2018-093548-B-I00"]}]},{"name":"Spanish Ministry of Science, Innovation and Universities","award":["Ram\u00f3n y Cajal: RYC-2016-20147"],"award-info":[{"award-number":["Ram\u00f3n y Cajal: RYC-2016-20147"]}]},{"name":"CONACYT fellowship Mexico","award":["283833"],"award-info":[{"award-number":["283833"]}]},{"name":"&quot;Doctorados Industriales 2018&quot; fellowship Comunidad de Madrid","award":["IND2018\/BIO-9610"],"award-info":[{"award-number":["IND2018\/BIO-9610"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Assisted reproductive technologies (ARTs) in the horse still yield suboptimal results in terms of pregnancy rates. One of the reasons for this is the lack of optimal conditions for the sperm capacitation in vitro. This study assesses the use of synthetic human tubal fluid (HTF) supplemented with D-penicillamine (HTF + PEN) for the in vitro capacitation of frozen\/thawed stallion spermatozoa by examining capacitation-related events over 180 min of incubation. Besides these events, we explored the in vitro capacity of the spermatozoa to migrate by thermotaxis and give rise to a population of high-quality spermatozoa. We found that HTF induced higher levels of hyperactive-like motility and protein tyrosine phosphorylation (PTP) compared to the use of a medium commonly used in this species (Whitten\u2019s). Also, HTF + PEN was able to maintain this hyperactive-like motility, otherwise lost in the absence of PEN, for 180 min, and also allowed for sperm selection by thermotaxis in vitro. Remarkably, the selected fraction was enriched in spermatozoa showing PTP along the whole flagellum and lower levels of DNA fragmentation when compared to the unselected fraction (38% \u00b1 11% vs 4.4% \u00b1 1.1% and 4.2% \u00b1 0.4% vs 11% \u00b1 2% respectively, t-test p &lt; 0.003, n = 6). This procedure of in vitro capacitation of frozen\/thawed stallion spermatozoa in HTF + PEN followed by in vitro sperm selection by thermotaxis represents a promising sperm preparation strategy for in vitro fertilization and intracytoplasmic sperm injection in this species.<\/jats:p>","DOI":"10.3390\/ani10091467","type":"journal-article","created":{"date-parts":[[2020,8,21]],"date-time":"2020-08-21T09:21:51Z","timestamp":1598001711000},"page":"1467","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["The Presence of D-Penicillamine during the In Vitro Capacitation of Stallion Spermatozoa Prolongs Hyperactive-Like Motility and Allows for Sperm Selection by Thermotaxis"],"prefix":"10.3390","volume":"10","author":[{"given":"Sara","family":"Ruiz-D\u00edaz","sequence":"first","affiliation":[{"name":"Department of Animal Reproduction, National Institute for Agriculture and Food Research and Technology (INIA), 28040 Madrid, Spain"},{"name":"Mistral Fertility Clinics S.L., Cl\u00ednica Tambre, 28002 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3547-3751","authenticated-orcid":false,"given":"Ivan","family":"Oseguera-L\u00f3pez","sequence":"additional","affiliation":[{"name":"Unidad Iztapalapa, Universidad Aut\u00f3noma Metropolitana, Ciudad de M\u00e9xico 09340, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9545-9230","authenticated-orcid":false,"given":"David","family":"De La Cuesta-D\u00edaz","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction, National Institute for Agriculture and Food Research and Technology (INIA), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bel\u00e9n","family":"Garc\u00eda-L\u00f3pez","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction, National Institute for Agriculture and Food Research and Technology (INIA), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Consuelo","family":"Serres","sequence":"additional","affiliation":[{"name":"Departamento de Medicina y Cirug\u00eda Animal, Facultad de Veterinaria, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Jos\u00e9","family":"Sanchez-Calabuig","sequence":"additional","affiliation":[{"name":"Departamento de Medicina y Cirug\u00eda Animal, Facultad de Veterinaria, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9893-9179","authenticated-orcid":false,"given":"Alfonso","family":"Guti\u00e9rrez-Ad\u00e1n","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction, National Institute for Agriculture and Food Research and Technology (INIA), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Serafin","family":"Perez-Cerezales","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction, National Institute for Agriculture and Food Research and Technology (INIA), 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1530\/REP-16-0420","article-title":"Why doesn\u2019t conventional IVF work in the horse? 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