Effect of antioxidants during bovine in vitro fertilization procedures on spermatozoa and embryo development.
Increased amounts of reactive oxygen species (ROS) during in vitro fertilization (IVF) may cause cytotoxic damage to gametes, whereas small amounts of ROS favour sperm capacitation. The aim of this study was to investigate the effect of antioxidants [50 microm beta-mercaptoethanol (beta-ME) and 50 microm cysteamine (Cyst)] or a pro-oxidant (5 mm buthionine sulfoximine) on the quality and penetrability of spermatozoa into bovine oocytes and on the subsequent embryo development and quality when added during IVF. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes, and mitochondrial function, was diminished (p < 0.05) after 4-h culture in the presence of antioxidants. Oocyte penetration rates were similar between treatments (p > 0.05), but antioxidants adversely affected the normal pronuclear formation rates (p < 0.05). The incidence of polyspermy was high for beta-ME (p < 0.05). No differences were observed in cleavage rates between treatments (p > 0.05). However, the developmental rate to the blastocyst stage was adversely affected by Cyst treatment (p < 0.05). The quality of embryos that reached the blastocyst stage, evaluated by total, inner cell mass (ICM) and trophectoderm cell numbers and ICM/total cell ratio was unaffected (p > 0.05) by treatments. The results indicate that ROS play a role in the fertilizing capacity in bovine spermatozoa, as well as in the interaction between the spermatozoa and the oocytes. It can be concluded that supplementation with antioxidants during IVF procedures impairs sperm quality, normal pronuclear formation and embryo development to the blastocyst stage.
Gonçalves
F S
FS
Department of Animal Reproduction, School of Agricultural and Veterinary Sciences, University of São Paulo State, Jaboticabal.
Barretto
L S S
LS
Arruda
R P
RP
Perri
S H V
SH
Mingoti
G Z
GZ
D000177:Acrosome / Q000187:drug effects; D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage*; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D019328:Buthionine Sulfoximine / Q000008:administration & dosage; D002417:Cattle; D002462:Cell Membrane / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure; D003543:Cysteamine / Q000008:administration & dosage; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects / Q000662:veterinary*; D008297:Male; D008623:Mercaptoethanol / Q000008:administration & dosage; D016877:Oxidants / Q000008:administration & dosage; D017382:Reactive Oxygen Species; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure
D000975:Antioxidants; D002843:Chromatin; D016877:Oxidants; D017382:Reactive Oxygen Species; D019328:Buthionine Sulfoximine; D003543:Cysteamine; D008623:Mercaptoethanol