Evaluating the impacts of osmotic and oxidative stress on common carp (Cyprinus carpio, L.) sperm caused by cryopreservation techniques.
Cryopreservation causes osmotic changes and oxidative damage that have sublethal and lethal effects on spermatozoa. We examined these osmotic and oxidative effects on common carp spermatozoa motility; membrane integrity; levels of thiobarbituric-acid-reactive substance (TBARS) and carbonyl groups (CP); and the activity of superoxide dismutase (SOD), glutathione reductase, and glutathione peroxidase (GPx). Sperm was diluted in dimethyl sulfoxide (DMSO) and ethylene glycol-based extenders, followed by equilibration, freezing, and thawing. Equilibration in DMSO extender resulted in a significant reduction of spermatozoa motility, but motility was induced in those spermatozoa following dilution with saline buffer, which usually inhibits undiluted spermatozoa motility. Spermatozoa velocity and membrane integrity decreased with both extenders following freezing and thawing. No significant difference in levels of TBARS or CP, or in SOD activity, was seen in samples equilibrated with either extender. The freeze/thaw process induced significantly higher levels of TBARS, CP, and GPx activity, but did not affect the level of SOD. Glutathione reductase activity was inhibited in samples exposed to DMSO extender. Ethylene glycol should be considered a preferred cryoprotective agent for common carp spermatozoa to reduce osmotic and oxidative stress during cryopreservation.
Li
Ping
P
Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic. pingli06@yahoo.com
Li
Zhi-Hua
ZH
Dzyuba
Boris
B
Hulak
Martin
M
Rodina
Marek
M
Linhart
Otomar
O
D000818:Animals; D017756:Aquaculture / Q000379:methods; D002347:Carps / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D019855:Ethylene Glycol / Q000494:pharmacology*; D029941:Fish Proteins / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D008297:Male; D018715:Microscopy, Video / Q000662:veterinary; D009997:Osmotic Pressure / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects*; D050050:Protein Carbonylation / Q000187:drug effects; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D003451:Cryoprotective Agents; D029941:Fish Proteins; D017392:Thiobarbituric Acid Reactive Substances; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D005980:Glutathione Reductase; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide