{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,4]],"date-time":"2026-01-04T23:19:24Z","timestamp":1767568764694,"version":"3.48.0"},"reference-count":57,"publisher":"Oxford University Press (OUP)","issue":"5","content-domain":{"domain":["rep.bioscientifica.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2025,9,17]]},"abstract":"<jats:sec>\n                    <jats:title>In brief<\/jats:title>\n                    <jats:p>Preservation of sperm, while successful in many species, can lead to molecular changes which make artificial insemination less successful. This study explores changes to proteasome function caused by chilling and freezing sperm, describing a new molecular change caused by sperm preservation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Abstract<\/jats:title>\n                    <jats:p>\n                      Sperm cryopreservation causes significant sublethal alterations, which ultimately impact\n                      <jats:italic>in vivo<\/jats:italic>\n                      fertility and reduce the efficacy of assisted reproductive technologies. A major driver of this damage is oxidative stress, which somatic cells combat with a coordinated cellular response, including actions of the proteasome. While sperm possess functional proteasomes with roles in capacitation and fertilisation, links between proteasomal proteolysis and cryopreservation-induced oxidative stress have never been explored. This study involved exposure of ram spermatozoa to cold shock (4\u00b0C, no cryoprotectant), chilled storage (24\u201348 h at 4\u00b0C with egg yolk), or cryopreservation compared to fresh controls. Direct impacts of oxidative stress were investigated by exposing fresh sperm to increasing levels of H\n                      <jats:sub>2<\/jats:sub>\n                      O\n                      <jats:sub>2<\/jats:sub>\n                      . Motility was assessed by computer-assisted sperm analysis, proteasome activities were measured using AMC-labelled oligopeptides, and proteasome subunit abundance was assessed by western blotting. Both caspase-like and chymotrypsin-like activities were not impacted by cold shock, significantly decreased in chilled samples, and significantly increased in frozen samples. Trypsin-like activity significantly decreased in cold shock and frozen samples, with a trend to decreasing in chilled samples. There was a weak positive correlation between trypsin-like activity and total motility (\n                      <jats:italic>r<\/jats:italic>\n                      <jats:sub>s<\/jats:sub>\n                      = 0.39,\n                      <jats:italic>P<\/jats:italic>\n                      = 0.001). All proteasome activities significantly decreased under oxidative stress. Proteasome subunit abundance showed an apparent increase following cryopreservation, but no change following chilled storage. This demonstrates that proteasome function may be altered by oxidative stress and chilled and frozen storage of spermatozoa. The functional implications of these alterations to sperm proteasome activity require further investigation.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1530\/rep-25-0008","type":"journal-article","created":{"date-parts":[[2025,9,17]],"date-time":"2025-09-17T13:48:15Z","timestamp":1758116895000},"update-policy":"https:\/\/doi.org\/10.1530\/crossmarkpolicy-11","source":"Crossref","is-referenced-by-count":0,"title":["Cold shock, chilling and cryopreservation impact sperm proteasome function"],"prefix":"10.1093","volume":"170","author":[{"given":"Grace","family":"Voss","sequence":"first","affiliation":[{"name":"School of Veterinary Science, The University of Queensland, Gatton, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mikayla","family":"Crouch","sequence":"additional","affiliation":[{"name":"School of Veterinary Science, The University of Queensland, Gatton, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2964-5940","authenticated-orcid":true,"given":"Michal","family":"Zigo","sequence":"additional","affiliation":[{"name":"Division of Animal Sciences, University of Missouri, Columbia, Missouri, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessica","family":"Epple-Farmer","sequence":"additional","affiliation":[{"name":"Division of Animal Science, Lincoln University of Missouri, Jefferson City, Missouri, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9231-2823","authenticated-orcid":true,"given":"Peter","family":"Sutovsky","sequence":"additional","affiliation":[{"name":"Division of Animal Sciences, University of Missouri, Columbia, Missouri, USA"},{"name":"Department of Obstetrics, Gynecology and Women\u2019s Health, University of Missouri, Columbia, Missouri, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4272-9943","authenticated-orcid":true,"given":"Taylor","family":"Pini","sequence":"additional","affiliation":[{"name":"School of Veterinary Science, The University of Queensland, Gatton, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","reference":[{"key":"bib1","series-title":"Mol Cell Proteomics","article-title":"Oxidative stress-mediated regulation of proteasome complexes","volume":"10","author":"Aiken CT","year":"2011","unstructured":"Aiken CT\n, \nKaake RM\n, \nWang X\n, et al.\n 2011 Oxidative stress-mediated regulation of proteasome complexes. 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