Effect of exogenous glutathione on the in vitro fertilization of bovine oocytes.
Increased amounts of reactive oxygen species (ROS) during in vitro culture may cause cytotoxic damage to gametes and embryos. The main purpose of this study was to investigate the effect of glutathione (GSH), a ROS scavenger, supplemented during IVF of bovine oocytes on embryo development using spermatozoa from different bulls. The following experiments were performed: 1) matured COCs were fertilized in the absence or presence of 1 mM GSH using semen from 4 bulls (Bulls A, B, C and D); 2) matured COCs were fertilized in the absence or presence of 1 mM GSH using semen from Bull C to examine sperm penetration, pronuclear formation and apposition; 3) COCs were fertilized with in the presence of either 0, 0.1, 1.0 or 10 mM GSH to examine the effect of GSH concentration using sperm from Bull C; 4) concentrations of GSH were measured both in the medium and in the oocytes during IVF. Glutathione at 1 mM in IVF medium affected the blastocyst formation, but not the cleavage rate. The effect on blastocyst formation was bull dependent: semen from Bull B and D had a negative, that from Bull C a positive and the one from Bull A no effect. The positive effect of Bull C semen increased the rate of blastocyst formation from 20.1 to 27.3% in control and GSH-treated samples, respectively. The increased rate was due to more zygotes reaching the 8-cell or greater stage by Day 4 after IVF. There was no change in the fertilization or cleavage rates. The GSH was still stable after 18 h incubation in IVF medium, and there was a dose-dependent increase in the GSH concentration in the oocytes. It is concluded that the effect of GSH during IVF on the proportion of blastocysts is dependent on both bull and GSH concentration.
Kim
I H
IH
Department of Farm Animal Health, Veterinary Faculty, Utrecht University, The Netherlands.
Van Langendonckt
A
A
Van Soom
A
A
Vanroose
G
G
Casi
A L
AL
Hendriksen
P J
PJ
Bevers
M M
MM
D000818:Animals; D002417:Cattle; D015925:Cryopreservation; D005314:Embryonic and Fetal Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D005978:Glutathione / Q000494:pharmacology*; D008297:Male; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000502:physiology; D012661:Semen / Q000166:cytology; D012662:Semen Preservation; D013084:Sperm-Ovum Interactions / Q000187:drug effects / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*
D017382:Reactive Oxygen Species; D005978:Glutathione