{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T08:57:09Z","timestamp":1785229029172,"version":"3.55.0"},"reference-count":61,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T00:00:00Z","timestamp":1777248000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T00:00:00Z","timestamp":1777248000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Andrology"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Challenges in establishing clinical conventional in vitro fertilization (IVF) in horses include variation in sperm and oocyte quality, which can be affected by mare age. Extended preincubation of stallion spermatozoa has resulted in successful IVF. However, there is limited research on the impact of preincubation on sperm characteristics and IVF success.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>Responses of frozen\u2013thawed stallion spermatozoa over time under IVF preincubation conditions were evaluated, and samples based on specific responses were tested for their ability to fertilize oocytes, including those from older mares, using conventional IVF.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Materials and Methods<\/jats:title>\n                    <jats:p>Frozen\u2013thawed spermatozoa were sorted, incubated for 10\u00a0h in capacitating conditions, and evaluated for motility, membrane integrity, mitochondrial function, and protein tyrosine phosphorylation (PY). Sperm samples for IVF were selected based on motion and PY patterns using oocytes from younger and older mares. Accessory spermatozoa binding to the zona pellucida (ZP), fertilization, cleavage, and blastocyst rates were assessed.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Sperm motility, velocities, linearity, and straightness were significantly reduced during incubation, while the amplitude of lateral head displacement increased. Plasma membrane integrity and mitochondrial activity were also significantly reduced over time. After 5\u00a0h of incubation, an increase in tyrosine\u2010phosphorylated proteins was observed, occurring primarily in the equatorial segment, but also including the midpiece or tail. The response of sperm samples to capacitating conditions varied. Sperm samples selected for IVF were successful at fertilizing oocytes from younger and older mares, with more accessory spermatozoa bound to ZP in oocytes from younger than older mares, although cleavage and blastocyst rates did not differ.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion and Conclusion<\/jats:title>\n                    <jats:p>We concluded that changes in sperm motion and PY patterns were indicative of hyperactivation and capacitation, although wide variation occurred among stallions. Cryopreserved spermatozoa can successfully fertilize oocytes using conventional IVF. Age of the mare oocyte donor does not appear to affect fertilization after IVF.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/andr.70249","type":"journal-article","created":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T05:39:34Z","timestamp":1777354774000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Hyperactivation and Tyrosine Phosphorylation of Cryopreserved Stallion Spermatozoa in Relation to Conventional In Vitro Fertilization Blastocyst Production"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6494-7557","authenticated-orcid":false,"given":"Raul A.","family":"Gonzalez\u2010Castro","sequence":"first","affiliation":[{"name":"Department of Biomedical Sciences Colorado State University  Fort Collins Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolina C.","family":"Porflidt","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences Colorado State University  Fort Collins Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra L.","family":"Ash","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences Colorado State University  Fort Collins Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacobo S.","family":"Rodriguez","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences Colorado State University  Fort Collins Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Perla D.","family":"Fleury","sequence":"additional","affiliation":[{"name":"In Vitro Equinos  Mogi Mirim S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward L.","family":"Squires","sequence":"additional","affiliation":[{"name":"In Vitro Equinos  Mogi Mirim S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elaine M.","family":"Carnevale","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences Colorado State University  Fort Collins Colorado USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,27]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.fertnstert.2024.03.003"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1017\/S1751731119002775"},{"key":"e_1_2_8_4_1","unstructured":"IETS \u201cStatistics of Embryo Production and Transfer in Domestic Farm Animals 2023 \u201d accessed March 10 2025 www.iets.org\/Committees\/Data\u2010Retrieval."},{"key":"e_1_2_8_5_1","first-page":"375","article-title":"In Vitro Fertilization in the Horse. 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