Fertilization Ability of Porcine Oocytes Reconstructed from Ooplasmic Fragments Produced and Characterized after Serial Centrifugations
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概要
- 論文の詳細を見る
Mitochondria are reported to be critical in in vitro 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 in vitro 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® 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 in vitro 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.
- 日本繁殖生物学会の論文
著者
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NOGUCHI Junko
Division of Agrobiological Resource II, National Institute of Agrobiological Resource
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Kikuchi Kazuhiro
Division Of Animal Sciences National Institute Of Agrobiological Sciences
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Nguyen Bui
Institute Of Biotechnology Vietnamese Academy Of Science And Technology
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Manabe Noboru
Animal Resource Center The University Of Tokyo:research Center. For Food Safety The University Of To
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Kaneko Hiroyuki
Division Of Animal Sciences National Institute Of Agrobiological Sciences
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Nakai Michiko
Division Of Animal Sciences National Institute Of Agrobiological Sciences
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Dang-nguyen Thanh
Institute Of Biotechnology Vietnamese Academy Of Science And Technology
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Viet Linh
Animal Resource Science Center Graduate School Of Agricultural And Life Sciences The University Of Tokyo
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NAGAI Takashi
NARO Institute of Livestock and Grassland Science
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SOMFAI Tamas
NARO Institute of Livestock and Grassland Science
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NGUYEN Bui
Institute of Biotechnology, Vietnamese Academy of Science and Technology, Hanoi, Vietnam
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VIET LINH
Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki 319-0206, Japan
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TANIHARA Fuminori
Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
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MEN Nguyen
Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
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VAN HANH
Institute of Biotechnology, Vietnamese Academy of Science and Technology, Hanoi, Vietnam
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MANABE Noboru
Animal Resource Science Center, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ibaraki 319-0206, Japan
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KANEKO Hiroyuki
Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
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KIKUCHI Kazuhiro
Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
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