{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T05:52:48Z","timestamp":1793944368872,"version":"4.8.0"},"reference-count":70,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2020,7,14]],"date-time":"2020-07-14T00:00:00Z","timestamp":1594684800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010198","name":"Ministerio de Econom\u00eda, Industria y Competitividad, Gobierno de Espa\u00f1a","doi-asserted-by":"publisher","award":["AGL2017-83098-R"],"award-info":[{"award-number":["AGL2017-83098-R"]}],"id":[{"id":"10.13039\/501100010198","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014180","name":"Junta de Castilla y Le\u00f3n","doi-asserted-by":"publisher","award":["JCyL-LE253P18"],"award-info":[{"award-number":["JCyL-LE253P18"]}],"id":[{"id":"10.13039\/501100014180","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>To improve artificial insemination protocols in ovine species it is crucial to optimize sperm quality evaluation after preservation technologies. Emerging technologies based on novel biomolecules and related to redox balance and proteins involved in sperm motility such as ProAKAP4 could be successfully applied in ram sperm evaluation. In this work, a multiparametric analysis of fresh, cooled, and cryopreserved ram sperm was performed at different complexity levels. Samples were evaluated in terms of motility (total motility, progressive motility, and curvilinear velocity), viability, apoptosis, content of reactive oxygen species, oxidation\u2012reduction potential, and ProAKAP4 expression and concentration. As expected, cryopreserved samples showed a significant decrease of sperm quality (p &lt; 0.05), evidencing different freezability classes among samples that were detected by ProAKAP4 analyses. However, in cooled sperm no differences were found concerning motility, viability, apoptosis, ROS content, and redox balance compared to fresh sperm that could explain the reported decrease in fertility rates. However, although the proportion of sperm ProAKAP4 positive-cells remained unaltered in cooled sperm compared to fresh control, the concentration of this protein significantly decreased (p &lt; 0.05) in cooled samples. This altered protein level could contribute to the decrease in fertility rates of cooled samples detected by some authors. More importantly, ProAKAP4 can be established as a promising diagnostic parameter of sperm quality allowing us to optimize sperm conservation protocols and finally improve artificial insemination in ovine species.<\/jats:p>","DOI":"10.3390\/biom10071046","type":"journal-article","created":{"date-parts":[[2020,7,20]],"date-time":"2020-07-20T10:59:38Z","timestamp":1595242778000},"page":"1046","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":52,"title":["ProAKAP4 as Novel Molecular Marker of Sperm Quality in Ram: An Integrative Study in Fresh, Cooled and Cryopreserved Sperm"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3655-6401","authenticated-orcid":false,"given":"Marta","family":"F. Riesco","sequence":"first","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Cellular Biology, Department of Molecular Biology, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7179-5742","authenticated-orcid":false,"given":"Luis","family":"Anel-Lopez","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Anatomy, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Neila-Montero","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Palacin-Martinez","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Montes-Garrido","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mercedes","family":"Alvarez","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2531-595X","authenticated-orcid":false,"given":"Paulino","family":"de Paz","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Cellular Biology, Department of Molecular Biology, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Anel","sequence":"additional","affiliation":[{"name":"Itra-ULE, INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1016\/j.theriogenology.2009.10.009","article-title":"Effect of storage duration, storage temperature, and diluent on the viability and fertility of fresh ram sperm","volume":"73","author":"Hanrahan","year":"2010","journal-title":"Theriogenology"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Kukovics, S., Gyoker, E., Nemeth, T., and Gergatz, E. 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