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2005 Fiscal Year Final Research Report Summary

Study of Japanese flounder cytokine gene promoter functions

Research Project

Project/Area Number 16580147
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General fisheries
Research InstitutionTokyo University of Marine Science and Technology

Principal Investigator

HIRONO Ikuo  Tokyo University of Marine Science and Technology, Course of Graduate School of Marine Science and Technology, Associate Professor, 海洋科学技術研究科, 助教授 (00270926)

Project Period (FY) 2004 – 2005
KeywordsJapanese Flounder / zebrafish / gene promoter / tumor necrosis factor / TNF / cell line / transgenic / cytokine
Research Abstract

Tumor necrosis factor (TNF) is known as a primary mediator of immune regulation and inflammatory response. However, little is known about TNF-mediated networks or signal cascades in fish. In this study, we characterized the regulation of gene expression of Japanese flounder TNF in cell lines and zebrafish embryo.
A recombinant plasmid containing the 2.4 kb promoter region of Japanese flounder tumor necrosis factor (TNF) and green fluorescence protein (GFP) was introduced into YO-K cells derived from Japanese flounder kidney, HINAE cells derived Japanese flounder embryos and zebrafish fertilized. YO-K cells and HINAE cells were incubated with lipopolysaccharide (LPS) at 20℃ for 24 h. The number of cells expressing GFP, as well as the amount of GFP protein was increased by LPS stimulation in both cell lines. GFP mRNA transcription was also induced by LPS stimulation in both YO-K cells and HINAE cells. In YO-K cells, expression level of GFP decreased gradually from 3 h to 6 h post stimulation, while a reverse trend was observed in HINAE cells. When the transgenic zebrafish embryos were exposed to LPS, GFP was expressed in the whole zebrafish embryonic surface and Western blot analysis also showed that the level of the expressed GFP protein in zebrafish embryo was higher at a LPS concentration of 140 μg/ml than at LPS concentrations of 0 and 70 μg/ml. These results indicate that transcription regulated by the Japanese flounder TNF promoter could be under the control of the LPS-recognition system in zebrafish embryos.

  • Research Products

    (4 results)

All 2005

All Journal Article (4 results)

  • [Journal Article] Functional analysis of tumor necrosis factor gene promoter from2005

    • Author(s)
      Yazawa R, Hirono I, Ohira T, Aoki T.
    • Journal Title

      Dev. Comp. Immunol 29

      Pages: 73-81

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Induction of Japanese flounder TNF promoter activity by lipopolysaccharide in zebra fish embryo.2005

    • Author(s)
      Yazawa R, Hirono I, Ohira T, Aoki T.
    • Journal Title

      Mar. Biotechnol., 7

      Pages: 231-235

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Induction of Japanese flounder TNF promoter activity by lipopolysaccharide in zebrafish embryo.2005

    • Author(s)
      Yazawa R, Hirono I, Ohira T, Aoki T.
    • Journal Title

      Marine Biotechnology 7

      Pages: 231-235

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Functional analysis of tumor necrosis factor gene promoter from Japanese flounder, Paralichthys olivaceus, using fish cell lines.2005

    • Author(s)
      Yazawa, R., Hirono, I., Ohira, T., Aoki, T.
    • Journal Title

      Development and Comparative Immunology 29

      Pages: 73-81

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2007-12-13  

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