Characterization of zebrafish Ikaros, a gene necessary for differentiation of the immune system.
| Project/Area Number |
14570783
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | The University of Tokyo (2003-2005)
Kansai Medical University (2002) |
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Principal Investigator |
KAWASAKI Hirohide The University of Tokyo, Institute of Medical Science, Instructor, 医科学研究所, 助手 (80278621)
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| 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)
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| Project Period (FY) |
2002 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| 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)
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| Keywords | Zebrafish / congenital bone marrow failure / congenital immunodificiency / in situ hybridization / ikaros / 先天性免疫不全 |
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| 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
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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
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Report
(5 results)
Research Products
(14 results)
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[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
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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
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