Effects of Cumulus Cells and Reactive Oxygen Species (ROS) on Plasminogen Activator Activity during In Vitro Maturation of Porcine Oocytes

  • Sa, Soo-Jin (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Park, Chun-Keun (College of Animal Life Science, Kangwon National University) ;
  • Kim, In-Cheul (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Lee, Seung-Hoon (Swine Science Division, National Institute of Animal Science, RDA) ;
  • Kwon, Oh-Sub (Swine Science Division, National Institute of Animal Science, RDA) ;
  • 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) ;
  • Cheong, Hee-Tae (Veterinary Medicine, Kangwon National University) ;
  • Webb, Bob (School of Biosciences, University of Nottingham, Sutton Bonington Campus)
  • Received : 2010.08.10
  • Accepted : 2010.08.26
  • Published : 2010.09.30

Abstract

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. The PA/plasmin system has been associated with a number of physiological processes such as fibrinolysis, ovulation and fertilization. Although correlations have been reported between reactive oxygen species (ROS) and oocyte maturation, the relationship between PA activity and ROS is unknown. The present study was undertaken to determine the effects of cumulus cells on PA activity in matured porcine oocytes under xanthine (X)-xanthine oxidase (XO) system. When oocytes were matured under the X-XO system, the proportion of oocytes remaining GV stage was higher (p<0.05) in oocytes without cumulus cells. The incidence of degenerated oocytes was higher (p<0.05) in the X+XO ($11.1{\pm}6.1$ and $21.6{\pm}3.4%$) than in the control group ($2.9{\pm}1.8$ and $4.0{\pm}1.6%$). The proportion of TUNEL-positive oocytes and activity of caspase-3 were higher (p<0.05) in cumulus-free oocytes and oocytes exposed to ROS. Tissue-type plasminogen activator-plasminogen activator inhibitor (tPA-PAI) and tissue-type plasminogen activator (tPA) activity were detected in oocytes that were separated from cumulus-oocytes complexs (COCs) at 44 h of maturation culture, and only tPA was produced in oocytes that were denuded before the onset of maturation culture. On the other hand, the activities of PA were increased (p<0.05) when oocytes were cultured under the X-XO system. The higher activity of tPA was observed in denuded oocytes (DOs) underwent apoptotic changes by oxidative stress. In COCs, however, tPA-PAI as well as tPA activity was detected and apoptotic changes such as DNA cleavage or caspase-3 activation were not observed. These results suggest that tP A may be relevant to apoptotic cell death in porcine oocytes by oxidative stress.

Keywords

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