title: Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders.
abstract: In this study, total glutathione content was determined in human spermatozoa before and after cryopreservation. Total GSH in fresh semen was 4.47±0.46nmol/10(8) cells. Following semen cryopreservation, GSH decreased to 1.62±0.13nmol/10(8) cells, a 64% reduction (p<0.01). This decrease in GSH content was associated with a decrease in sperm progressive motility (68% of reduction, p<0.01). Addition of 1mM GSH to the freezing extender increased the percentage of total motility and sperm viability. It also modified the motility pattern measured by CASA with changes in the straight-line and average path velocities and wobble of the curvilinear trajectory. Addition of GSH to the freezing media reduced spermatozoa ROS levels and increased the level of sulfhydryl groups on membrane proteins. Nevertheless, no effect of GSH addition on lipid membrane disorder or chromatin condensation was detected. Addition of 1 or 5mM GSH to the thawing media increased the percentage of motile and progressively motile spermatozoa, but no effect on viability was detected. In conclusion, the antioxidant defensive capacity of the GSH is severely altered by the freeze-thawing process. The addition of GSH to the freezing and thawing extender could be of partial and limited benefit in improving the function of frozen human spermatozoa.
authors.affiliation: Dept. Physiology, University of Murcia, Murcia 30100, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Gadea
authors.forename: M
authors.initials: M
authors.lastname: Molla
authors.forename: E
authors.initials: E
authors.lastname: Selles
authors.forename: M A
authors.initials: MA
authors.lastname: Marco
authors.forename: F A
authors.initials: FA
authors.lastname: Garcia-Vazquez
authors.forename: J C
authors.initials: JC
authors.lastname: Gardon
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000378:metabolism / Q000494:pharmacology; D005615:Freezing / Q000009:adverse effects; D005978:Glutathione / Q000032:analysis* / Q000494:pharmacology*; D006801:Humans; D008297:Male; D017382:Reactive Oxygen Species / Q000032:analysis; D055101:Semen Analysis / Q000379:methods; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects*; D013094:Spermatozoa / Q000737:chemistry* / Q000187:drug effects* / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D003451:Cryoprotective Agents; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D005978:Glutathione