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Characterization of zebrafish Ikaros, a gene necessary for differentiation of the immune system.

Research Project

Project/Area Number14570783
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Pediatrics
Research InstitutionThe University of Tokyo(2003-2005)
Kansai Medical University(2002)

Principal Investigator

KAWASAKI Hirohide  The University of Tokyo, Institute of Medical Science, Instructor, 医科学研究所, 助手 (80278621)

Co-Investigator(Kenkyū-buntansha) 国府寺 美  関西医科大学, 医学部, 助手 (90198614)
中野 崇秀  関西医科大学, 医学部, 助手 (40333207)
Project Period (FY) 2002 – 2005
Project Status Completed(Fiscal Year 2005)
Budget Amount *help
¥2,800,000 (Direct Cost : ¥2,800,000)
Fiscal Year 2005 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 2004 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 2003 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 2002 : ¥1,000,000 (Direct Cost : ¥1,000,000)
KeywordsZebrafish / congenital bone marrow failure / congenital immunodificiency / in situ hybridization / ikaros / 先天性免疫不全
Research Abstract

Vertebrate hematopoietic cells are derived from self-renewing multipotential stem cells (HSCs) that are specified early during embryonic development. The lymphoid lineages are derived from the HSCs under the influence of transcription factors. One of these, Ikaros, a zinc finger-containing gene, is intimately involved in early regulation of lymphoid development of the T-, B- and NK-cell lineage and is also involved in immune function in adults. We employed RT-PCR and conserved primers to clone a zebrafish Ikaros fragment from adult spleen mRNA, then used this fragment to isolate a full-lemgth cDNA transcript from an adult spleen cDNA library. The nucleotide sequence of this full-length clone (2305bp) encodes a protein of 537 amino acids, including six consensus Drosophia kruppel-like (CX2CX12HX3-5H) zinc finger motifs. Alignment and phylogenetic analysis of all available Ikaros sequences are consistent with the predicted evolutionary conservation of a canalized regulator of lymphopoies … More is. Early developmental expression patterns have been analyzed by whole embryo in situ hybridization. At the 5 somite stage, Ikaros is expressed in dorsolateral mesodermal stripes in a pattern identical to early hematopoietic transcription factors, such as Gata-1 and SCL. At 24hourspost fertilization (hpf) there is strong expression of Ikaros in the ICM, the yolk sac equivalent in zebrafish and putative site of early HSC differentiation. At 46 hpf Ikaros expression is observed in the dorsal aorta, the AGM equivalent in zebrafish. Starting at 72 hpf the expression of Ikaros is restricted to two main area : the bilateral thymi and rostral CNS ganglia. This expression pattern persists throughout the period of observation (up to 7 days pf). Tissue-specific expression patterns and sequence characterization of alternative-spliced isoforms are currently being investigated. We have also isolated a contig of genomic PAC clones encoding zebrafish Ikaros and have constructed a partial restriction map of the region. Nucleotide sequences have been obtained from within and around this locus in order to determine intron-exon boundaries and for generating polymorphisms, the latter being used to localize the Ikaros gene to linkage group 13. A 130kb PAC done encoding the Ikaros genomic locus is presently being modified using a GFP reporter construct in order to make transgenic individuals for recapitulation and characterization of the in vivo expression patterns of this gene. With our interest in development of the adaptive immune system we have initiated an early pressure screen for identifying mutations affecting lymphoid ontogeny. Ikaros will be a very useful marker to characterize the identified lymphoid mutants. Less

Report

(5results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report

Research Products

(14results)

All 2005 2004 2002 2001 Other

All Journal Article Publications

  • [Journal Article] RAS-blocking bisphosphonate zoledronic acid inhibits the abnormal proliferation and differentiation of juvenile myelomonocytic leukemia cells in vitro.2005

    • Author(s)
      Yoshitoshi Ohtsuka, Atsushi Manage, Hirohide Kawasaki et al.
    • Journal Title

      Blood 106

      Pages : 3134-3141

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Methylation statusof the P15 and p16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia.2005

    • Author(s)
      Daisuke Hasegawa, Atsushi Manabe, Takeo Kubota, Hirohide Kawasaki et al.
    • Journal Title

      British Journal of Haematology 128

      Pages : 805-812

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] RAS-blocking bisphosphonate zoledronic acid inhibits the abnormal proliferation and differentiation of juvenile myelomonocytic leukemia cells in vitro.2005

    • Author(s)
      Yoshitoshi Ohtsuka, Atsushi Manabe, Hirohide Kawasaki, Daisuke Hasegawa, Yuji Zaike, Sumiko Watanabe, Takakuni Tanizawa, Tatsutoshi Nakahata, Kohichiro Tsuji
    • Journal Title

      Blood 106

      Pages : 3134-3141

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Methylation status of the p15 and p16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia.2005

    • Author(s)
      Hasegawa, Atsushi Manabe, Takeo Kubota, Hirohide Kawasaki, Imiko Hirose, Yoshitoshi Ohtsuka, Toshihisa Tsuruta, Yasuhiro Ebihara, Yu-ichi Goto, Xiao Yan Zhao, Kazuo Sakashita, Kenichi Koike, Mariko Isomura, Seiji Kojima, Akinori Hoshika, Kohichiro Tsuji, Tatsutoshi Nakahata
    • Journal Title

      British Journal of Haematology 128(6)

      Pages : 805-812

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] RAS-blocking bisphosphonate zoledronic acid inhibits the abnormal proliferation and differentiation of juvenile myelomonocytic leukemia cells in vitro.2005

    • Author(s)
      Ohtsuka Y, Manabe A, Kawasaki H, et al.
    • Journal Title

      Blood 印刷中

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Methylation status of the p15 and p16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia.2005

    • Author(s)
      Hasegawa D, Manabe A, Kubota T, Kawasaki H, et al.
    • Journal Title

      British Journal of Haematology 128・6

      Pages : 805-812

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Methylation status of the p15 and p16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia.2005

    • Author(s)
      Hasegawa D, Manabe A, Kubota T, Kawasaki H, et al.
    • Journal Title

      British Journal of Haematology (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] 後天性サイトメガロ感染症に合併した免疫学的血小板減少性紫斑病2004

    • Author(s)
      河崎裕英, 他
    • Journal Title

      日本小児血液学会雑誌 18・5

      Pages : 561-565

    • Related Report
      2005 Annual Research Report 2004 Annual Research Report
  • [Journal Article] CD7/CD13陽性急性分類不能型白血病2004

    • Author(s)
      長谷川大輔, 真部淳, 鶴田敏久, 大塚欣敏, 海老原康博, 河崎裕英, 他
    • Journal Title

      日本小児血液学会雑誌 18・3

      Pages : 155-159

    • Related Report
      2005 Annual Research Report 2004 Annual Research Report
  • [Journal Article] The effect of thrombopoietin on erythroid progenitors in Diamond-Blackfan anemia.2002

    • Author(s)
      Hirohide Kawasaki, et al.
    • Journal Title

      International Journal of Hematology 75・4

      Pages : 382-384

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] The effect of thrombopoietin on erythroid progenitors in Diamond-Blackfan anemia.2002

    • Author(s)
      Hirohide Kawasaki, Takahide Nakano, Urara Kohdera, Yohnosuke Kobayashi
    • Journal Title

      International Journal of Hematology 75(4)

      Pages : 382-384

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Ikaros expression as a marker for lymphoid progenitors during zebrafish development.2001

    • Author(s)
      Willett CE, Kawasaki H, et al.
    • Journal Title

      Developmental Dynamics 222・4

      Pages : 694-698

    • Related Report
      2005 Annual Research Report
  • [Publications] Hirohide Kawasaki, et al.: "The effect of thrombopoietin on erythroid progenitors in Diamond-Blackfan anemia."International Journal of Hematology. 75・4. 382-384 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hirohide Kawasaki, et al.: "The effect of thrombopoietin on erythroid progenitors in Diamond-Blackfan anemia"International Journal of Hematology. 75・4. 382-384 (2002)

    • Related Report
      2002 Annual Research Report

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Published : 2002-04-01   Modified : 2016-04-21  

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