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Effect of plasmin on sperm penetration and zona pellucida solubility in pigs
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/362133
In our previous study, exogenous plasminasminogen activators (PAs) influenced to fertility of boar spermatozoa via reduction of zona pellucida (ZP) resistance against protease and number of sperm binding ZP. plasminasmin (plasmin), is converted by PAs, is an important enzyme to degrade extracellular matrix and it is closely associated with fertilization process. Therefore, the aim of present study was to confirm changes of sperm penatration and ZP solubility by plasmin during in vitro fertilization (IVF). The cumulus-oocyte complasminexes (COCs) were aspirated from the antral follicles 3-6 mm in diameter and matured for 44 hours. Then, the cumulus cells were removed and denuded oocytes were co-incubated with spermatozoa for 18-20 hours in IVF medium containing 100 ng/ml plasmin. The number of sperm binding ZP and ZP solubility were measured using hoechst 33342 and 0.5% (w/v) pronase, respectively. Aceto-orcein stain was used to assess fertilization parameters. In results, sperm penetration did not affect by plasmin treatment during fertilization. Hoewever, treatment of plasmin decreased monospermic fertilization and IVF efficiency compared with control group (p<0.05). Furthermore, the number of penetrated sperm and pronucleus formation per zygote in plasmin group was significantly increased compared with control group (p<0.05). Despite of reduced monospermic fertilization by plasmin treatment, the number of sperm binding ZP was significantly higher in non-treated zygote than plasmin-treated zygote (p<0.05). Similar with previous study, ZP digestion time was reduced by plasmin treatment (p<0.05). These findings shown that plasminasmin during fertilization enhance the penetration of spermatozoa into ZP via increasing of ZP solubility and it was correspond with our previous results that fertility of spermatozoa during IVF was increased by exogenous urokinase-type PA treatment via sperm-ZP binding and increase of ZP solubility. Therefore, during the fertilization process, plasmin that is converted by PAs from oviduct epithelial cells might be closely associated with degradation of ZP proteins for penetration of sperm. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).
