Enhancement of developmental competence after in vitro fertilization of porcine oocytes by treatment with ascorbic acid 2-O-alpha-glucoside during in vitro maturation.
The present study was conducted to examine the effect of ascorbic acid 2-O-alpha-glucoside (AA-2G), a stable ascorbate derivative, on the sustenance of cytoplasmic maturation responsible for subsequent developmental competence after in vitro fertilization of porcine oocytes. Cumulus-oocyte complexes were cultured for 44 h in North Carolina State University 37 medium supplemented with cysteine, gonadotropins, 10% (v:v) porcine follicular fluid, and 0-750 microM AA-2G. When oocytes were matured in the presence of 250 microM AA-2G, their ability to promote transformation of the sperm nucleus into the male pronucleus (MPN) was strongly enhanced after in vitro fertilization. Similarly, the presence of 25 microM beta-mercaptoethanol (ME) enhanced the degree of progression to MPN of penetrated sperm by associating with the increase in intracellular glutathione (GSH) content. Although the AA-2G treatment during oocyte maturation showed no influence on the GSH concentration, significantly higher levels of ascorbic acid (AsA) were detected in these oocytes than in those oocytes cultured without AA-2G (P < 0.05). The length of DNA migration encompassed by reactive oxygen species (ROS), generated by the hypoxanthine-xanthine oxidase system, was not increased in the oocytes treated with AA-2G, whereas ME treatment could not block the DNA damage by ROS. These findings indicate that AA-2G in maturation medium can potentiate the cellular protection of oocytes against oxidative stress by continuously supplying AsA. The proportion of development to the blastocyst stage after in vitro insemination was significantly increased in oocytes matured with AA-2G (P < 0.05), and this proportion showed no difference in comparison with that of oocytes treated with ME. These findings suggest that a critical concentration of intracellular AsA, supplied by AA-2G during in vitro maturation, plays an important role in supporting the cytoplasmic maturation responsible for developmental competence after fertilization by prevention of oxidative stress against porcine oocytes.
Tatemoto
H
H
School of Bioresources, Hiroshima Prefectural University, Shobara, Hiroshima 727-0023, Japan. hidettmt@bio.hiroshima-pu.ac.jp
Ootaki
K
K
Shigeta
K
K
Muto
N
N
D000818:Animals; D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000494:pharmacology*; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology; D046508:Culture Techniques; D004249:DNA Damage / Q000187:drug effects; D004622:Embryo, Mammalian / Q000737:chemistry / Q000502:physiology*; D005314:Embryonic and Fetal Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects; D005978:Glutathione / Q000032:analysis; D019271:Hypoxanthine / Q000378:metabolism; D008623:Mercaptoethanol / Q000494:pharmacology; D009865:Oocytes / Q000502:physiology*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D013552:Swine; D014969:Xanthine Oxidase / Q000378:metabolism
D017382:Reactive Oxygen Species; D019271:Hypoxanthine; C065378:ascorbic acid 2-O-glucoside; D008623:Mercaptoethanol; D014969:Xanthine Oxidase; D005978:Glutathione; D001205:Ascorbic Acid