{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T10:18:43Z","timestamp":1784715523247,"version":"3.55.0"},"reference-count":50,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2003,5,29]],"date-time":"2003-05-29T00:00:00Z","timestamp":1054166400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Reproduction Devel"],"published-print":{"date-parts":[[2003,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Fluo\u20104 loaded immature oocytes were cooled from 30\u00b0C to various lower temperatures between 20 and 10\u00b0C and changes in intracellular calcium (Ca<jats:sup>2+<\/jats:sup>) levels were measured. Pig oocytes cooled to 14\u00b0C exhibited a clear biphasic Ca<jats:sup>2+<\/jats:sup> rise. Lower temperatures produced similar responses, while higher temperatures did not exert any effect. The Ca<jats:sup>2+<\/jats:sup> response appeared to rely on ryanodine dependent stores as removal of extracellular Ca<jats:sup>2+<\/jats:sup> and intracytoplasmic injection of heparin did not modify cold\u2010induced Ca<jats:sup>2+<\/jats:sup> elevation, while procaine or ruthenium red virtually eliminated the response. Confocal analysis of subcellular Ca<jats:sup>2+<\/jats:sup> distribution during cooling revealed that the ion rises sharply within the nucleus. As Ca<jats:sup>2+<\/jats:sup> imbalance may activate nuclear endonucleases, DNA integrity of cooled pig oocytes was evaluated by TUNEL and comet assays. Most cooled oocytes showed clear signs of DNA fragmentation. Oocytes injected with 1,2\u2010bis(2\u2010aminophenoxy)ethane\u2010<jats:italic>N,N,N<\/jats:italic>\u2032<jats:italic>,N<\/jats:italic>\u2032\u2010tetracetic acid tetrapotassium salt (BAPTA), a Ca<jats:sup>2+<\/jats:sup> chelator, maintained their DNA integrity thus confirming that intracellular Ca<jats:sup>2+<\/jats:sup> is involved in triggering DNA fragmentation. The protective effect exerted by ruthenium red and\/or procaine further confirmed this hypothesis. These data show that a moderate and transient cooling is sufficient to cause an intracellular Ca<jats:sup>2+<\/jats:sup> rise that leads to DNA damage. The addition of inhibitors of ryanodine dependent Ca<jats:sup>2+<\/jats:sup> stores may represent a valuable protective treatment to reduce chilling injuries. Mol. Reprod. Dev. 65: 289\u2013297, 2003. \u00a9 2003 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/mrd.10275","type":"journal-article","created":{"date-parts":[[2003,5,30]],"date-time":"2003-05-30T10:06:10Z","timestamp":1054289170000},"page":"289-297","source":"Crossref","is-referenced-by-count":34,"title":["Cold\u2010induced calcium elevation triggers DNA fragmentation in immature pig 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