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Role of transcription factor, NF-kB in preimplantation development and zygotic gene activation in mice

Research Project

Project/Area Number 13660287
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied animal science
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

YAMADA Masayasu  Kyoto Univ. Grad. School of Agri., Associate Professor, 農学研究科, 助教授 (10243073)

Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2001: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsNF-kB / Mouse embryo / Nuclear localization / DNA binding / Gel shift assay / Antisense oligmer / Preimplantation development / P65-p50 heyerodimer / 免疫蛍光染色 / 核内への局在 / RelA-p50二量体
Research Abstract

Nuclear facotr kappa B (NF-kB) is a transcription factor and was originally described as being present in the cytosol of most most cell types as an inactive heterodimer composed of 50-kDa (p50) and 65-kDa (p65) subunits and bound to one of the IkB inhibitory proteins. After degrdation of IkB by its phosphorylation, NF-kB is activated and then p50 and p65 heterodimer translocates to the nucleus, where it directly binds its cognate DNA sequence. It has recently been found that NF-kB plays a pivotal role in morphogenesis and embryonic development, such as the formation of Drosophila ventral structures and chicken limb buds. However, the role of NF-kB in preimplantation development in mammals has not yet been clarified. In this study, the expression and the activation of NF-kB during preimplantation development in mice were first examined. Immunocytochemical studies using anti-p65 peptide monoclonal antibody showed intense cytoplasmic staining in embryos at the 1-cell to the blastocyst sta … More ges and that nuclear localization of p65, which is indicative of an active form of NF-kB was observed in 1-cell and 2-cell embryos. Gel shift analysis revealed that the DNA binding activity of NF-kB was found in embrys at the 1-cell and 2-cell stages, and the supershift analysis using anti-p50 and p65 antibodies confirmed that the active NF-kB in mouse 1-cell and 2-cell embryos is composed of p50 and p65 heterodimer. In the second experiments, effects of phosphorothioate antisense oligonucleotide (ODN) against p65 on in vitro development in mice were examined. Treatments with antisense ODN p65 of 1-cell embryos induced their developmental arrest at the 2-cell or 4-cell stages, whereas sense ODN p65 had no effects on their development. When embryos at the 2-cell stage were treated with antisense or sense ODN p65 , most embryos developed to the blastocyst stage. These results suggest that NF-kB plays an important role for regulation of development of preimplantation embryos. Furthermore, the identification of specific target genes under transcriptional regulation by NF-kB will definitely enhance our understanding of gene expression and development of preimplantation. Less

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Nishikimi A.: "Localization of nitric oxide synthase activity in unfertilized oocytes and fertilized embryos during preimplantation development in mice"Reproduction. 122. 957-963 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nishikimi A.: "Nuclear translocation of a pre-mRNA splicing factor, p100^<prpl/zer1/prp6>, in mouse 1-cell embryos"J. Reprod. Dev.. 48. 257-263 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Matsukawa T.: "Alleviation of the two-cell block of ICR mouse embryos by polyaminocarboxylate metal chelators"Reproduction. 124. 65-71 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kurosaka S.: "Dependence of DNA synthesis and in vitro development of bovine nuclear transfer embryos on the stage of the cell cycle of donor cells and recipient cytoplasts"Biol. Reprod.. 67. 643-647 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Azuma T.: "Effects of EDTA saturated with Ca2+ (Ca-EDTA) on pig, bovine and mouse oocytes at the germinal vesicle stage during maturation culture and the involvement of chelation of Zn2+ in pronuclear formation induction by Ca-EDTA"Reproduction. 124. 235-240 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ikeda S.: "Apoptosis in cumulus cells during in vitro maturation of bovine cumulus-enclosed oocytes"Reproduction. 125. 369-376 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nishikimi A., et al.: "Localization of nitric oxide synthase activity in unfertilized oocytes and fertilized embryos during preimplantation development in mice"Reproduction. 122. 957-963 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nishikimi A., et al.: "Nuclear translocation of a pre-mRNA splicing factor, p100^<*****>, in mouse 1-cell embryos"J. Reprod Dev.. 48. 257-263 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Matsukawa T.: "Alleviation of the two-cell block of ICR mouse embryos by polyaminocarboxylate metal chelatars."Reproduction. 124. 65-71 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kurosaka s.: "Dependence of DNA synthesis and in vitro development of bovine nuclear transfer embryos on the stage of the cell cycle of donor cells and recipient cytoplasts."Biol. Reprod.. 67. 643-647 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Azuma T.: "Effects of EDTA saturated with Ca2+ (Ca-EDTA) on pig, bovine and mouse oocytes at the germinal vesicle stage during maturation culture and the involvement of chelation of Zn2+ in pronuclear formation induction by Ca-EDTA"Reproduction. 124. 235-240 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ikeda S.: "Apoptosis in cumulus cells during in vitro maturation of bovine cumulus-enclosed oocytes"Reproduction. 125. 369-376 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kurosaka S.: "Dependence of DNA synthesis and in vitro development of bovine nuclear transfer embryos on the stage of the cell cycle of donor cells and recipient cytoplasts"Biol. Reprod.. 67. 643-647 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Azuma T.: "Effects of EDTA saturated with Ca2+ (Ca-EDTA) on pig, bovine and mouse oocytes at the germinal vesicle stage during maturation culture and the involvement of chelation of Zn2+in pronuclear formation induction by Ca-EDTA."Reproduction. 124. 235-240 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ikeda S.: "Apoptosis in cumulus cells during in vitro maturation of bovine cumulus-enclosed oocytes"Reproduction. 125. 369-376 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nishikimi A.: "Localization of nitric oxide synthase activity in unfertilized oocytes and fertilized embryos during preimplantation development in mice"Reproduction. 122. 957-963 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nishikimi A.: "Nuclear translocation of a pre-mRNA splicing factor, p100^<prp1/zer1/prp6>, in mouse 1-cell embryos"J. Reprod. Dev.. (印刷中).

    • Related Report
      2001 Annual Research Report
  • [Publications] Matsukawa T.: "Alleviation of the two-cell block of ICR mouse embryos by polyaminocarboxylate metal chelators"Reproduction. (印刷中).

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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