Effects of oxygen concentration and antioxidants on the in vitro developmental ability, production of reactive oxygen species (ROS), and DNA fragmentation in porcine embryos.
After in vitro maturation and fertilization of porcine oocytes, the fertilized embryos were cultured under 5 or 20% oxygen (O2) for 7 days. In embryos cultured under 5% O2 versus 20% O2, development to the blastocyst stage was higher (36.3% versus 22.5%, P < 0.05); the hydrogen peroxide (H2O2) content as a reactive oxygen species was lower (92 pixels versus 111 pixels, P < 0.05); and fragmentation of DNA in 8- to 16-cell stage embryos (estimated by the comet assay) resulted in a shorter (P < 0.05) DNA tail (36 microm versus 141 microm). Antioxidants such as beta-mercaptoethanol (beta-ME) and Vitamin-E (Vit-E) suppressed oxidative damage in the embryos and improved their developmental ability. For embryos cultured under 20% O2, there were the following differences (P < 0.05) between embryos exposed to 0 microM versus 50 microM beta-ME: 28% versus 57% developed to the blastocyst stage; 125 pixels versus 98 pixels per embryo in H2O2 content; and a DNA tail of 209 microm versus 105 microm. In addition, for embryos cultured under 20% O2, there were also differences (P < 0.05) between those exposed to 0 microM versus 50 microM of Vit-E: 28% versus 40% rate of development to the blastocyst stage; 28.9 cells versus 35.9 cells in the expanded blastocyst; 122 pixels versus 95 pixels per embryo (H2O2 content); and 215 microm versus 97 microm length of the DNA tail. Therefore, a low O2 concentration during in vitro culture of porcine embryos decreased the H2O2 content and, as a consequence, reduced DNA fragmentation, and, thereby, improved developmental ability.
Laboratory of Animal Breeding and Reproduction, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Yuichiro
Y
Kitagawa
Keita
K
Suzuki
Akihiro
A
Yoneda
Tomomasa
T
Watanabe
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002970:Cleavage Stage, Ovum; D020552:Comet Assay; D003543:Cysteamine / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D010100:Oxygen / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013552:Swine / Q000196:embryology*; D014810:Vitamin E / Q000494:pharmacology
D000975:Antioxidants; D017382:Reactive Oxygen Species; D014810:Vitamin E; D003543:Cysteamine; D006861:Hydrogen Peroxide; D010100:Oxygen