{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T04:59:50Z","timestamp":1790398790084,"version":"4.1.0"},"reference-count":26,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T00:00:00Z","timestamp":1789776000000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP24K18026"],"award-info":[{"award-number":["JP24K18026"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Reprod Domestic Animals"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Toll\u2010like receptor 2 (TLR2) has been implicated in fertilization\u2010related functions of bovine spermatozoa, but post\u2010thaw responses to pharmacological TLR1\/2 modulation across kinematic and membrane\u2010related characteristics remain less defined. One ejaculate from each of four Holstein bulls was used and semen was exposed after thawing to a TLR1\/2 agonist, a TLR1\/2 antagonist or no additive. Five independent experimental repetitions per bull were performed using separate straws from the same ejaculate; therefore, the biological donor sample size was four bulls. Computer\u2010assisted sperm analysis and\n                    <jats:italic>k<\/jats:italic>\n                    \u2010means clustering identified six kinematically defined motility subpopulations and flow cytometry assessed plasma membrane and acrosomal integrity, membrane fluidity and mitochondrial superoxide\u2010related fluorescence using MitoSOX Red. At 3\u2009h, antagonist exposure reduced total motility compared with control and agonist treatments (30.1%\u2009\u00b1\u20096.3% vs. 40.4%\u2009\u00b1\u20095.6% and 37.7%\u2009\u00b1\u20095.8%, respectively; adjusted\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.001). Conversely, antagonist\u2010treated samples contained more spermatozoa with intact plasma membrane and acrosome than agonist\u2010treated samples (52.4%\u2009\u00b1\u20093.4% vs. 48.4%\u2009\u00b1\u20093.1%; adjusted\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.001) and more viable spermatozoa with low MitoSOX fluorescence (31.1%\u2009\u00b1\u20096.1% vs. 22.4%\u2009\u00b1\u20094.7%; adjusted\n                    <jats:italic>p<\/jats:italic>\n                    \u2009=\u20090.001). Agonist exposure increased the proportion of viable spermatozoa with high membrane fluidity and was associated with higher MitoSOX fluorescence than antagonist exposure. These findings describe distinct post\u2010thaw functional profiles following pharmacological TLR1\/2 agonist and antagonist exposure; they do not establish TLR2\u2010specific causality or effects on fertilizing competence.\n                  <\/jats:p>","DOI":"10.1111\/rda.70321","type":"journal-article","created":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T07:37:21Z","timestamp":1789803441000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effects of Post\u2010Thaw Toll\u2010Like Receptor 1\/2 Agonist and Antagonist Exposure on Motility Subpopulations and Functional Characteristics of Bovine Spermatozoa"],"prefix":"10.1111","volume":"61","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7277-5970","authenticated-orcid":false,"given":"Chihiro","family":"Kanno","sequence":"first","affiliation":[{"name":"Laboratory of Clinical Veterinary Medicine for Large Animals, School of Veterinary Medicine Kitasato University  Towada Aomori Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toju","family":"Oshima","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Veterinary Medicine for Large Animals, School of Veterinary Medicine Kitasato University  Towada Aomori Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1236-3461","authenticated-orcid":false,"given":"Shogo","family":"Sato","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Veterinary Medicine for Large Animals, School of Veterinary Medicine Kitasato University  Towada Aomori Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5059-3164","authenticated-orcid":false,"given":"Yosuke","family":"Maeda","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Veterinary Medicine for Large Animals, School of Veterinary Medicine Kitasato University  Towada Aomori Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-6453-8718","authenticated-orcid":false,"given":"Fumiaki","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Veterinary Medicine for Large Animals, School of Veterinary Medicine Kitasato University  Towada Aomori 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