{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T17:45:33Z","timestamp":1784569533094,"version":"3.55.0"},"reference-count":73,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2024,10,16]],"date-time":"2024-10-16T00:00:00Z","timestamp":1729036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005357","name":"Slovak Research and Development Agency","doi-asserted-by":"publisher","award":["APVV-21-0095"],"award-info":[{"award-number":["APVV-21-0095"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005357","name":"Slovak Research and Development Agency","doi-asserted-by":"publisher","award":["APVV-21-0168"],"award-info":[{"award-number":["APVV-21-0168"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005357","name":"Slovak Research and Development Agency","doi-asserted-by":"publisher","award":["1\/0067\/24"],"award-info":[{"award-number":["1\/0067\/24"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005357","name":"Slovak Research and Development Agency","doi-asserted-by":"publisher","award":["NUKLEUS 313011V387"],"award-info":[{"award-number":["NUKLEUS 313011V387"]}],"id":[{"id":"10.13039\/501100005357","id-type":"DOI","asserted-by":"publisher"}]},{"name":"VEGA","award":["APVV-21-0095"],"award-info":[{"award-number":["APVV-21-0095"]}]},{"name":"VEGA","award":["APVV-21-0168"],"award-info":[{"award-number":["APVV-21-0168"]}]},{"name":"VEGA","award":["1\/0067\/24"],"award-info":[{"award-number":["1\/0067\/24"]}]},{"name":"VEGA","award":["NUKLEUS 313011V387"],"award-info":[{"award-number":["NUKLEUS 313011V387"]}]},{"name":"Integrated Infrastructure","award":["APVV-21-0095"],"award-info":[{"award-number":["APVV-21-0095"]}]},{"name":"Integrated Infrastructure","award":["APVV-21-0168"],"award-info":[{"award-number":["APVV-21-0168"]}]},{"name":"Integrated Infrastructure","award":["1\/0067\/24"],"award-info":[{"award-number":["1\/0067\/24"]}]},{"name":"Integrated Infrastructure","award":["NUKLEUS 313011V387"],"award-info":[{"award-number":["NUKLEUS 313011V387"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Theobromine (TBR) is a methylxanthine known for its bronchodilatory and stimulatory effects. This research evaluated the vitality, capacitation patterns, oxidative characteristics, microbial profile and expression of capacitation-associated proteins (CatSper1\/2, sodium bicarbonate cotransporter [NBC], protein kinases A [PKA] and C [PKC] and adenylate cyclase 10 [ADCY10]) in cryopreserved bovine spermatozoa (n = 30) in the absence (cryopreserved control [CtrlC]) or presence of different TBR concentrations (12.5, 25, and 50 \u00b5M) in egg yolk extender. Fresh ejaculate served as a negative control (CtrlN). Significant post-thaw maintenance of the sperm motility, membrane and DNA integrity and mitochondrial activity (p &lt; 0.001) were recorded following the administration of 25 \u03bcM and 50 \u03bcM TBR, then compared to CtrlC. All groups supplemented with TBR exhibited a significantly lower percentage of prematurely capacitated spermatozoa (p &lt; 0.001) than CtrlC. Significantly decreased levels of global reactive oxygen species (ROS), hydrogen peroxide and hydroxyl radicals were observed in the presence of 25 \u03bcM and 50 \u03bcM TBR (p &lt; 0.01). Western blot analysis revealed that supplementation with 50 \u03bcM TBR significantly prevented the loss of NBC and ADCY10 (p &lt; 0.01), while all TBR doses stabilized the levels of PKC (p &lt; 0.05 at 50 \u03bcM TBR; p &lt; 0.001 at 12.5 \u03bcM and 25 \u03bcM TBR). In summary, we suggest that TBR is effective in protecting the spermatozoa during the cryopreservation process through its potential to stimulate energy synthesis while preventing ROS overproduction and the loss of proteins involved in the sperm activation process.<\/jats:p>","DOI":"10.3390\/cells13201710","type":"journal-article","created":{"date-parts":[[2024,10,16]],"date-time":"2024-10-16T10:11:04Z","timestamp":1729073464000},"page":"1710","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Cryoprotective Potential of Theobromine in the Improvement of the Post-Thaw Quality of Bovine Spermatozoa"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9652-0446","authenticated-orcid":false,"given":"Filip","family":"Benko","sequence":"first","affiliation":[{"name":"Institute of Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u0160tefan","family":"Ba\u0148as","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2907-5909","authenticated-orcid":false,"given":"Michal","family":"\u010eura\u010dka","sequence":"additional","affiliation":[{"name":"AgroBioTech Research Centre, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4460-0222","authenticated-orcid":false,"given":"Miroslava","family":"Ka\u010d\u00e1niov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Horticulture, Faculty of Horticulture and Landscape Engineering, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia"},{"name":"School of Medical and Health Sciences, University of Economics and Human Sciences in Warsaw, Okopowa 59, 010 43 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2895-1249","authenticated-orcid":false,"given":"Eva","family":"Tvrd\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ugur, M.R., Abdelrahman, A.S., Evans, H.C., Gilmore, A.A., Hitit, M., Arififiantini, R.I., Purwantara, B., Kaya, A., and Memili, E. (2019). Advances in Cryopreservation of Bull Sperm. Front. Vet. 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