Evaluation of Zona Pellucida Function for Sperm Penetration During In Vitro Fertilization in Pigs

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  • TANIHARA Fuminori
    The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
  • NAKAI Michiko
    Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
  • KANEKO Hiroyuki
    Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
  • NOGUCHI Junko
    Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
  • OTOI Takeshige
    The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan
  • KIKUCHI Kazuhiro
    The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan

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  • Evaluation of Zona Pellucida Function for Sperm Penetration During <i>In Vitro</i> Fertilization in Pigs

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In porcine oocytes, the function of the zona pellucida (ZP) with regard to sperm penetration or prevention of polyspermy is not well understood. In the present study, we investigated the effects of the ZP on sperm penetration during in vitro fertilization (IVF). We collected in vitro-matured oocytes with a first polar body (ZP+ oocytes). Some of them were freed from the ZP (ZP− oocytes) by two treatments (pronase and mechanical pipetting), and the effects of these treatments on sperm penetration parameters (sperm penetration rate and numbers of penetrated sperm per oocyte) were evaluated. There was no evident difference in the parameters between the two groups. Secondly, we compared the sperm penetration parameters of ZP+ and ZP− oocytes using frozen-thawed epididymal spermatozoa from four boars. Sperm penetration into ZP+ oocytes was found to be accelerated relative to ZP− oocytes. Thirdly, we evaluated the sperm penetration of ZP+ and ZP− oocytes at 1−10 h after IVF (3 h gamete co-incubation). The proportions of oocytes penetrated by sperm increased significantly with time in both groups; however, the number of penetrated sperm per oocyte did not increase in ZP− oocytes. Finally, we performed IVF using ZP− oocytes divided into control (3 h) and prolonged gamete co-incubation (5 h) groups. Greater numbers of sperm penetrated in the 5 h group than in the control group. These results suggest that the ZP and oolemma are not competent factors for prevention of polyspermy in our present porcine IVF system. However, it appears that ZP removal is one of the possibilities for reducing polyspermic penetration in vitro in pigs.

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