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Semen taken from each of the seven Akkaraman rams were pooled. Semen collection was done twice a week and it continued for 3\u2009weeks. Each pooled semen sample was divided into six equal groups and diluted with tris\u2009+\u2009egg yolk extender including 0 (control), 200, 400, 800\u2009nM, 1 and 5\u2009\u03bcM concentrations of C<jats:sub>60<\/jats:sub>HyFn at 37\u00b0C. They were then frozen in liquid nitrogen vapour at \u2212140\u00b0C, stored in liquid nitrogen container (\u2212196\u00b0C) and thawed at 37\u00b0C for 25\u2009s before analysis. In comparison with control, C<jats:sub>60<\/jats:sub>HyFn addition prior to freezing procedure provided significant increases in total and progressive motility rates, glutathione peroxidase, catalase activities and percentage of highly active mitochondria, and significant decreases in dead and abnormal sperm rates, lipid peroxidation, caspase\u20103 and DNA fragmentation levels in frozen\u2013thawed ram semen. When compared to control, C<jats:sub>60<\/jats:sub>HyFn supplementation significantly down\u2010regulated the expression levels of miR\u2010200a and KCNJ11, and significantly up\u2010regulated the expression levels of miR\u20103958\u20103p (at the concentrations of 200, 400, 800\u2009nM and 1\u2009\u03bcM), CatSper1 (at the concentrations of 200, 400\u2009nM and 5\u2009\u03bcM), CatSper2 (at the concentrations of 1 and 5\u2009\u03bcM), CatSper3 (at the concentrations of 200, 400\u2009nM, 1 and 5\u2009\u03bcM), CatSper4 (at all concentrations), ANO1 (at the concentrations of 800\u2009nM, 1 and 5\u2009\u03bcM) and TRPV5 (at the concentrations of 200, 400 and 800\u2009nM). The addition of C<jats:sub>60<\/jats:sub>HyFn had no effect on global DNA methylation rates. As a result, C<jats:sub>60<\/jats:sub>HyFn supplementation to ram semen extenders may be beneficial in reducing some of the functional, structural and molecular damages in sperm induced by the freeze\u2013thawing procedure.<\/jats:p>","DOI":"10.1111\/rda.14513","type":"journal-article","created":{"date-parts":[[2023,12,1]],"date-time":"2023-12-01T17:03:43Z","timestamp":1701450223000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Reduction of cryopreservation\u2010induced structural, functional and molecular damages in ram sperm by hydrated C<sub>60<\/sub> fullerene"],"prefix":"10.1111","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5250-1478","authenticated-orcid":false,"given":"\u0130brahim Halil","family":"G\u00fcng\u00f6r","sequence":"first","affiliation":[{"name":"Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine F\u0131rat University  Elaz\u0131\u011f 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