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Relationship between In Vitro Maturation and Plasminogen Activator Activity on Porcine Cumulus-Oocytes Complexes Exposed to Oxidative Stress

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한국동물번식학회 (The Korean Society of Animal Reproduction)
초록

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 (H2O2), the proportion of the germinal vesicle breakdown (GVBD) and oocytes maturation were decreased with addition of H2O2, and were significantly (p<0.05) lower in medium with 0.1 mM H2O2 than control group. Also, the rate of degenerated oocytes was increased in as H2O2 concentration increased. 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 H2O2 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 H2O2 and (3) control medium with 1.0 mM H2O2 along with catalase in concentrations of 0.01, 0.1, and 1.0 mg/ml, respectively. H2O2 decreased the rate of GVBD and maturation in porcine COCs but catalase revealed protective activity against oxidative stress caused by H2O2. In this experiment, tPA and tPA-PAI activities were higher in media with 1.0 mM H2O2 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.

목차
ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    Culture Media    Preparation of Oocytes    Experimental Design    SDS-PAGE and Zymography    Statistics   RESULTS   DISCUSSION   REFERENCES
저자
  • Soo Jin Sa(Swine Science Division, National Institute of Animal Science, RDA)
  • Chun Keun Park(College of Animal Life Science, Kangwon National University)
  • Hee Tae Cheong(Veterinary Medicine, Kangwon National University)
  • Jung Ho Son(Noah Biotech. Inc.)
  • Myung Jick Kim(Swine Science Division, National Institute of Animal Science, RDA)
  • Kyu Ho Cho(Swine Science Division, National Institute of Animal Science, RDA)
  • Du Wan Kim(Swine Science Division, National Institute of Animal Science, RDA)
  • Kyoung Min So(Swine Science Division, National Institute of Animal Science, RDA)
  • In Cheul Kim(Swine Science Division, National Institute of Animal Science, RDA) Corresponding author