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Basic study for the production of nock-out pig by nucleiar transfer using recombinant somatic cell

Research Project

Project/Area Number 12480248
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory animal science
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

MORI Tadashi  Hokkaido Univ., Grad.School of Agriculture, Inst., 大学院・農学研究科, 助手 (30230072)

Co-Investigator(Kenkyū-buntansha) TAKAHASHI Yoshiyuki  Hokkaido Univ., Grad.School of Veterinary Med., Prof., 大学院・獣医学研究科, 教授 (70167485)
MATSUMOTO Kenichi  Hokkaido Univ., Grad.School of Pharm.Sci., Asso.Prof., 大学院・薬学研究科, 助教授 (30202328)
YOSHIDA Ikuya  Hokkaido Univ., Center of Advan Sci.and Tech., Inst., 先端科学技術研究センター, 助手 (90240275)
SUZUKI Keita  Hokkaido Univ., Field Scientific Center, Inst., 北方生物圏フィールド科学センター, 助手 (60261335)
Project Period (FY) 2000 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥8,100,000 (Direct Cost: ¥8,100,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsPig / Oocytes / Embryos / IP3 / Ca^<2+> concentration / PhospholipaseCζ / Nuclear transfer / EGFP gene / 核移植胚 / 遺伝子発現 / ジーンターゲッティング / Xist / 遺伝子導入 / 核初期化 / ゲノミックインプリティング / ゲノミックインプリンティング / ビエゾドライブマイクロマニュビレーター / カルシウムオシレーション
Research Abstract

Inositol 1,4,5-trisphosphate (IP3) is considered to be important for activation of mammalian oocytes at the time of fertilization, and that the activation induces a rise in intracellular Ca^<2+> concentration. Therefore, IP3 can be useful as an artificial activator of oocytes. In this study, we injected IP3 into porcine matured oocytes and demonstrated that injected IP3 promotes the development of the matured oocytes and that their developmental ability is positively correlated with the rise in Ca^<2+> concentration induced by IP3. Naturally, mammalian oocytes are activated with sperm penetration, and it is suggested that some unknown sperm factor must activate the oocytes. For several years, it has been argued what is a real sperm factor to activate oocytes. Recently, phospholipaseC(PLC)ζ cDNA is cloned and is expected to be the most useful candidate to activate the oocytes. We originally succeed to clone the porcine PLCζcDNA. In addition to these activation works, and finally in vitro developmental study of porcine nuclear transfer embryos from foreign gene-transfected somatic cells was investigated. We used enhanced green fluorescent protein(EGFP) gene as a foreign gene maker. The result indicated that EGFP gene transfected into oocytes has no deleterious effect in the embryonic development to blastocyst stage. However, the clear transfer fetus which is expressing EGFP gene could not be obtained so far.

Report

(5 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • 2000 Annual Research Report

Research Products

(20 results)

All 2005 2004 2003 2002 2000 Other

All Journal Article (12 results) Publications (8 results)

  • [Journal Article] Roles of cumulus cells during maturation of porcine in the rise in intracellular Ca2+ induced by inositol 1,4,5-triphosphate.2005

    • Author(s)
      Amano T., Mori T.et al.
    • Journal Title

      Theriogenology (In press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Activation and development of porcine oocyte maturated in vitro following injection of inositol 1.4.5-triphosphate.2004

    • Author(s)
      Amano T., Mori T.et al.
    • Journal Title

      Animal Reproduction Science 80

      Pages: 101-112

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Effects of delipidaton and oxygen concentration on in vitro development of porcine embryos.2004

    • Author(s)
      Yoneda A., Morf T.et al.
    • Journal Title

      Journal of Reproduction and Development 50

      Pages: 287-295

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Activation and development of porcine oocyte maturated in vitro following injection of inositol 1,4,5-triphosphate.2004

    • Author(s)
      Amano T., Mori T., Watanabe T.
    • Journal Title

      Animal Reproduction Science 80

      Pages: 101-112

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Effects of delipidation and oxgen concentration on in vitro development of porcine embryos.2004

    • Author(s)
      Yoneda A., Suzuki K., Mori T., Ueda J., Watanabe T.
    • Journal Title

      Journal of Reproduction and Development 50

      Pages: 287-295

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Effect of fusion/activation prptocol on in vitro development of porcine nuclear transfer embryos constructed with foreign gene-transfected fetal fibroblasts.2003

    • Author(s)
      Matinez Diaz M.A., Mori T.et al.
    • Journal Title

      Journal of Veterinary Medical Science 65

      Pages: 989-994

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Effect of fusion/activation protocol on in vitro development of porcine nuclear transfer embryos constructed with foreign gene-transfected fetal fibroblasts.2003

    • Author(s)
      Martinez Diaz M.A., Mori T., Nagano M., Katagiri S., Shimizu H.
    • Journal Title

      Journal of Veterinary Medical Science 65

      Pages: 989-994

    • NAID

      110003886296

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] The suppression of fragmentation by stabilization of actin filament in porcine enucleated oocytes.2002

    • Author(s)
      Kawahara M., Mori T.et al.
    • Journal Title

      Theriogenology 58

      Pages: 1081-1095

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] The supression of fragmentation by stabilization of actin filament in porcine enucleated oocytes.2002

    • Author(s)
      Kawahara M., Mori T., Tanaka H., Shimizu H.
    • Journal Title

      Theriogenology 58

      Pages: 1081-1095

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Roles of gap junctional communication of cumulus cells in cytoplasmic maturation of porcine oocytes.2000

    • Author(s)
      Mori T., Shimizu H.et al.
    • Journal Title

      Biology of Reproduction 62

      Pages: 913-919

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Roles of gap junctional communication of cumulus cells in cytoplasmic maturation of porcine oocytes.2000

    • Author(s)
      Mori T., Amano T., Shimizu H.
    • Journal Title

      Biology or Reproduction 62

      Pages: 913-919

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Journal Article] Role of cumulus cells during maturation of porcine oocyte in the rise in intracellular Ca^<2+> induced by inositol 1,4,5-triphosphate.

    • Author(s)
      Amano T., Mori T., Watanabe T.
    • Journal Title

      Theriogenology (In press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Manabu Kawahara: "The suppression of fragmentation by stablization of actin filament in porcine enucleated oocytes"Theriogenology. 58. 1081-1095 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.A.Martinez Diaz: "Effects of cycloheximide treatment and interval between fusion and activation on in vitro development of pig nuclear transfer embryos"Reprod Fertil Dev. 14. 191-197 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Koji Ikeda: "Effects of butyrolactonr I on the inhibition of meiotic resumption of porcine oocytes and subsequent development after somatic cell nuclear transfer"J Mamm Ova Res. 19. 46-51 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tadashi Mori: "Estradiol production by porcine parthenogenetic embryos during early pregnancy"Theriogenology. 59. 371 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Manabu Kawahara: "The supressiom of fragmentation by stabilization of actin filament in porcine encleated oocytes"Teriogenology. 58. 1081-1095 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.A.Martinez Diaz: "Effects of cycloheximide treatment and interval between fusion and activation in vitro development of pig nuclear transfer embryos"Reprod Fertil Dev. 14. 191-197 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Koji Ikeda: "Effects of butyrolactone I on the inhibition of meiotic resumption of porcine oocytes and subsequent development after somatic cell nuclear tranfer"J Mamm Ova Res. 19. 46-51 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tadashi Mori: "Estradiol production by porcine parthenogenetic embryos during early pregnancy"Teriogenology. 59. 371-371 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2000-03-31   Modified: 2016-04-21  

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