Fertilization Ability of Porcine Oocytes Reconstructed from Ooplasmic Fragments Produced and Characterized after Serial Centrifugations

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Author(s)

    • VIET LINH Nguyen VIET LINH Nguyen
    • Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki 319-0206, Japan|Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan|Institute of Biotechnology, Vietnamese Academy of Science and Technology, Hanoi, Vietnam
    • KIKUCHI Kazuhiro SOMFAI Tamas
    • NARO Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), Ibaraki 305-0901, Japan
    • NAGAI Takashi
    • NARO Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), Ibaraki 305-0901, Japan
    • MANABE Noboru
    • Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki 319-0206, Japan
    • KIKUCHI Kazuhiro
    • Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan|The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan
    • NAKAI Michiko
    • Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
    • TANIHARA Fuminori
    • Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan|The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan
    • NOGUCHI Junko
    • 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
    • DANG-NGUYEN Thanh Quang
    • Institute of Biotechnology, Vietnamese Academy of Science and Technology, Hanoi, Vietnam|Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba 305-8572, Japan|NARO Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), Ibaraki 305-0901, Japan
    • MEN Nguyen Thi
    • Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan|Institute of Biotechnology, Vietnamese Academy of Science and Technology, Hanoi, Vietnam|NARO Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), Ibaraki 305-0901, Japan
    • VAN HANH Nguyen
    • Institute of Biotechnology, Vietnamese Academy of Science and Technology, Hanoi, Vietnam

Abstract

Mitochondria are reported to be critical in <i>in vitro</i> maturation of oocytes and subsequent embryo development after fertilization, but their contribution for fertilization has not been investigated in detail. In the present study, we investigate the contribution of mitochondria to fertilization using reconstructed porcine oocytes by fusion of ooplasmic fragments produced by serial centrifugations (centri-fusion). Firstly, we evaluated the characteristics of ooplasmic fragments. Three types of fragments were obtained by centrifugation of porcine oocytes matured <i>in vitro</i> for 46 h: brownish (B), transparent (T) and large (L) fragments containing both B and T parts in a fragment. The production efficiencies of these types of fragments were 71.7, 91.0 and 17.8 fragments/100 oocytes, respectively. In experiments, L fragments were excluded because they contained both brownish and transparent components that were apparently intermediate between B and T fragments. Observations by confocal microscopy after staining with MitoTracker Red CMXRos<sup>®</sup> and transmission electron microscopy revealed highly condensed active mitochondria in B fragments in contrast to T fragments that contained only sparse organelles. We reconstructed oocytes by fusion of a karyoplast and two cytoplasts from B and T fragments (B and T oocytes, respectively). The B oocytes showed higher sperm penetration (95.8%) and male pronuclear formation rates (94.2%) by <i>in vitro</i> fertilization than T oocytes (66.7% and 50.0%, respectively). These results suggest that the active mitochondria in oocytes may be related to their ability for fertilization.

Journal

  • Journal of Reproduction and Development

    Journal of Reproduction and Development 59(6), 549-556, 2013

    THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

Codes

  • NII Article ID (NAID)
    130003361058
  • NII NACSIS-CAT ID (NCID)
    AA10936678
  • Text Lang
    ENG
  • ISSN
    0916-8818
  • NDL Article ID
    025084146
  • NDL Call No.
    Z54-H305
  • Data Source
    NDL  J-STAGE 
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