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Effect of Exogenous Plasmiongen Activators and Their Inhibitor on In Vitro Fertilization in Pigs

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The aim of present study was to confirm changes of results in in vitro fertilization (IVF) by two types of exogenous plasminogen activators (urokinase-type, uPA; tissue- type, tPA), and their inhibitor (PAI-1) and investigate the regulatory mechanism. the cumulus-oocyte complexes (COCs) were aspirated from 3~6 mm antral follicles and matured for 44 hours. Then, the cumulus cells were removed for fertilization and denuded oocytes were co-incubated with spermatozoa for 18~20 hours in IVF medium containing 100 pg/mL uPA, tPA or PAI-1. The number of sperm that bound to zona pellucida (ZP) and ZP solubility were measured using hoechst 33342 and 0.5% (w/v) pronase, respectively. Aceto-orcein stain was used to assess sperm penetration and sperm characteristics were analyzed by flow cytometry. In result, sperm penetration was increased by uPA and PAI-1 treatment than other groups (p<0.05). Interestingly, treatment of tPA enhanced ratio of monopsermic fertilization in penetrated zygotes population and reduced polyspermy than uPA groups (p<0.05). Similar to sperm penetration, number of penetrated sperm per zygote were higher PAI-1 treatment than tPA group (p<0.05). The number of ZP bounded sperm and ZP digestion time were lower in uPA and tPA-treated zygotes than other treatment groups (p<0.05) and ZP resistance was increased by treatment of PAI-1 (p<0.05). In results in sperm assay, acrosome reaction did not affected by all treatment in live and all sperm population. However, tPA treatment increased ratio of damaged sperm (p<0.05). These findings shown that exogenous PAs could affect to result of IVF via regulation ZP resistance, ZP binding, and sperm viability and two-types PAs differently regulated the fertility of boar spermatozoa.

저자정보

  • Yong Hwangbo College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea
  • Ji-Eun Lee College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea
  • Hee-Tae Cheong College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea
  • Boo-Keun Yang College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea
  • Choon-Keun Park College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea

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