{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T15:47:13Z","timestamp":1753890433181,"version":"3.41.2"},"reference-count":37,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,5,16]],"date-time":"2025-05-16T00:00:00Z","timestamp":1747353600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The livestock industry has witnessed a notable increase in the proportion of male calves born following artificial insemination with cryopreserved semen, hinting at a possible association between cryotolerance and inherent structural or functional disparities between X- and Y-chromosome bearing sperm. To delve into this phenomenon, we conducted a comprehensive proteomic analysis on bovine sex-sorted semen.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Sperm samples were meticulously categorized based on stringent quality parameters. Concurrent sorting of X- and Y-sperm from identical ejaculates was performed, followed by protein extraction for subsequent analysis. Quantitative isobaric tags for relative and absolute quantification (iTRAQ) were employed to identify differentially expressed proteins.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>iTRAQ pinpointed 20 proteins with differential expression between X- and Y-spermatozoa, including 12 upregulated and 8 downregulated proteins in Y-sperm. Aquaporin 7 (AQP7), significantly upregulated in Y-sperm, was identified as a key candidate. Western blot analysis corroborated elevated AQP7 expression in Y-sperm compared to X-sperm, while Aquaporin 3 (AQP3) showed no significant difference. Immunofluorescence revealed AQP7 localization to the post-acrosomal region, midpiece, and principal piece of Y-sperm with higher fluorescence intensity, whereas AQP3 distribution was comparable between groups.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>These findings imply a potential role for AQP7 in augmenting Y-sperm cryotolerance, providing molecular insights into sex-specific cryopreservation effects on fertility outcomes. Future research should elucidate AQP7\u2019s functional mechanisms and explore implications for livestock breeding technologies.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcell.2025.1582961","type":"journal-article","created":{"date-parts":[[2025,5,16]],"date-time":"2025-05-16T09:00:32Z","timestamp":1747386032000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Comparative analysis of AQP7 expression and cryotolerance in X- and Y-chromosome bearing bovine sperm"],"prefix":"10.3389","volume":"13","author":[{"given":"Jieru","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruchun","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fei","family":"Huang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peng","family":"Niu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanling","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dongfang","family":"Yuan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan","family":"Du","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Congcong","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mingcai","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiajia","family":"Suo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qinghua","family":"Gao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,5,16]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.7176\/JBAH\/9-1-01","article-title":"Calves sex ratio in artificial and naturally bred cattle at Adami Tulu agricultural research center","volume":"9","author":"Arega","year":"2019","journal-title":"J. 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