Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Relationship between In Vitro Maturation and Plasminogen Activator Activity on Porcine Cumulus-Oocytes Complexes Exposed to Oxidative Stress

  • Sa, Soo-Jin (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Park, Chun-Keun (College of Animal Life Science, Kangwon National University) ;
  • Cheong, Hee-Tae (Veterinary Medicine, Kangwon National University) ;
  • Son, Jung-Ho (Noah Biotech. Inc.) ;
  • Kim, Myung-Jick (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Cho, Kyu-Ho (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Kim, Du-Wan (Swine Science Division, National Institute of Animal Science, RDA) ;
  • So, Kyoung-Min (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Kim, In-Cheul (Swine Science Division, National Institute of Animal Science, RDA)
  • Received : 2011.08.29
  • Accepted : 2011.09.05
  • Published : 2011.09.30
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Abstract

This study was undertaken to evaluate the relationship between in vitro maturation and plasminogen activators (PAs) activity on porcine cumulus-oocytes complexes (COCs) exposed to oxidative stress. When COCs were cultured in maturation medium with hydrogen peroxide ($H_2O_2$), the proportion of the germinal vesicle breakdown (GVBD) and oocytes maturation were decrease with addition of $H_2O_2$, and were significantly (p<0.05) lower in medium with 0.1 mM $H_2O_2$ than control group. Also, the rate of degenerated oocytes was increased in as $H_2O_2$ concentration in eased. When COCs were cultured for 48 h, three plasminogen-dependent lytic bands were observed: tissue-type PA (tPA); urokinase-type PA (uPA); and tPA-PA inhibitor (tPA-PAI). PA activity was quantified using SDS-PAGE and zymography. When $H_2O_2$ concentration was increased, tPA and tPA-PAI activities also increased in porcine oocytes cultured for 48 h, but not uPA. In other experiment, embryos were divided into three groups and cultured in (1) control medium, (2) control medium with 1.0 mM $H_2O_2$ and (3) control medium with 1.0 mM $H_2O_2$ along with catalase in concentrations of 0.01, 0.1, and 1.0 mg/ml, respectively. $H_2O_2$ decreased the rate of GVBD and maturation in porcine COCs but catalase revealed protective activity, against oxidative stress caused by $H_2O_2$. In this experiment, tPA and tPA-PAI activities were higher in media with 1.0 mM $H_2O_2$ alone. Increasing concentration of catalase decreased tPA and tPA-PAI activities in porcine oocytes. These results indicate that the exposure of porcine follicular oocytes to ROS inhibits oocytes maturation to metaphase-II stage and increase the oocytes degeneration. Also, we speculated that increased ROS level may trigger tPA and tPA-PAI activities in porcine oocytes matured in vitro.

Keywords

References

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