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Analysis of molecules involved in mouse primordial germ cell development using transgenic mice

Research Project

Project/Area Number 11670211
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Experimental pathology
Research InstitutionOsaka University

Principal Investigator

KIMURA Tohru  Research Institute for Micobial Diseases, Osaka University, Research Associate, 微生物病研究所, 助手 (50280962)

Co-Investigator(Kenkyū-buntansha) NAKANO Toru  Research Institute for Micobial Diseases, Osaka University, Professor, 微生物病研究所, 教授 (00172370)
伊川 正人  大阪大学, 遺伝情報実験施設, 助手 (20304066)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,700,000 (Direct Cost: ¥2,700,000)
KeywordsGerm lineage / primordial germ cells / TNAP gene / Oct-3 / 4 gene / transgenic mice / EGFP / Cre / loxP system / loxP系 / 生殖質 / germ cell-less(gcl)遺伝子
Research Abstract

Primordial germ cells (PGC) are founder cells of all gametes. Because of the absence of efficient experimental system, it remains largely unknown what kind of molecular mechanisms are operating during the commitment and maintenance of mouse germ lineage. In order to establish the experimental system to investigate the PGC development, we generated the following transgenic mice using the promoters of TNAP (tissue nonspecific alkaline phosphatase) and Oct-3/4 genes, the marker genes for PGC.
(1) We established the mice expressing fluorescent protein EGFP in PGC by knocking-in EGFP cDNA into TNAP locus. In vivo visualization of PGC was successfully achieved in this mouse. Furthermore, live PGC can be isolated at 100% purity using cell sorter. These convenient visualization and purification techniques should be useful for analyzing PGC development at cellular and molecular levels.
(2) We tried to generate a model mouse for PGC-specific gene expression system. For this purpose, we produced the transgenic mouse containing the transgene in which floxed-EGFP cDNA followed by _-galactosidase cDNA is under the control of Oct-3/4 gene promoter. As expected, EGFP but not _-galactosidase was expressed in PGC of the transgenic mice. On the other hand, it had been expected that expression of _-galactosidase would have turned on in PGC when this mouse was crossed with transgenic mouse expressing Cre recombinase in PGC.However, _-galactosidase activity was not detected in PGC.The failure to detect the transgene expression is probably due to the excesives loss of transgenes that resulted from the recombination between loxP sites of tandemly inserted transgenes.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Fukushi M,Dixon J,Kimura T,Tsurutani N,Dixon MJ,Yamamoto N.: "Identification and cloning of a novel cellular protein Naf1, Nef-associated factor 1, that increases cell surface CD4 expression."FEBS Letters. 442. 83-88 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Suzuki Y,Koyanagi Y,Tanaka Y,Murakami T,Misawa N, et al.: "Determinant in human immunodeficiency virus type 1 for efficient replication under cytokine-induced CD4+ T-helper1 (Th1)-and Th2-type conditions."J.Virol.. 73. 316-324 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Motoyama N,Kimura T,Takahashi T,Watanabe T,Nakano T.: "bcl-x prevents apoptotic cell death of both primitive and definitive erythrocytes at the end of maturation."J.Exp.Med.. 189. 1691-1698 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kimura T,Yomogida K,Iwai N,Kato Y,Nakano T.: "Molecular cloning and genomic organization of mouse homologue of Drosophila germ cell-less and its expression in germ lineage cells."Biochem.Biophys.Res.Commun.. 262. 223-230 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kusunoki A,Wada A,Kurosaki N,Kimura T,Takai K,Yamamoto N, et al.: "Antisense oligodeoxynucleotide complementary to CXCR4 mRNA block replication of HIV-1 in COS cells."Nucleosides Nucleotides. 18. 1705-1708 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Kimura,Y.Sonoda,N.T.Izui,M.Suda,K.Sasaki,T.Nakano: "Proliferation and cell death of embryonic primitive erythrocytes."Exp.Hematol.. 28. 635-641 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fukushi M, Dixon J, Kimura T, Tsurutani N, Dixon MJ, and Yamamoto N.: "Identification and cloning of a novel cellular protein Naf1, Nef-associated factor 1, that increases cell surface CD4 expression."FEBS Letters. 442. 83-88 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Suzuki Y, Koyanagi Y, Tanaka Y, Murakami T, Misawa N, Maeda N, Kimura T, Shida H, Hoxie JA, O'Brien WA, and Yamamoto N.: "Determinant in human immunodeficiency virus type 1 for efficient replication under cytokine-induced CD4+ T-helper 1 (Th1)- and Th2-type conditions."J.Virol.. 73. 316-324 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Motoyama N, Kimura T, Takahashi T, Watanabe T, and Nakano T.: "bcl-x prevents apoptotic cell death of both primitive and definitive erythrocytes at the end of maturation."J.Exp.Med.. 189. 1691-1698 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kimura T, Yomogida K, Iwai N, Kato Y, and Nakano T.: "Molecular cloning and genomic organization of mouse homologue of Drosophila germ cell-less and its expression in germ lineage cells."Biochem.Biophys.Res.Commun.. 262. 223-230 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kusunoki A, Wada A, Kurosaki N, Kimura T, Takai K, Yamamoto N, and Takaku H.: "Antisense oligodeoxynucleotide complementary to CXCR4 mRNA block replication of HIV-1 in COS cells."Nucleosides Nucleotides. 18. 1705-1708 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kimura T, Sonoda Y, Iwai N, Satoh M, Yamaguchi-Tsukio M, Izui T, Suda M, Sasaki K, and Nakano T.: "Proliferation and cell death of embryonic primitive erythrocytes."Exp.Hematol.. 28. 635-641 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Kimura,Y.Sonoda,N.T.Izui,M.Suda,K.Sasaki,T.Nakano: "Proliferation and cell death of embryonic primitive erythrocytes"Exp Hematol. 28. 635-641 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] M.Fukushi et al.: "Identification and cloning of a novel cellular protein Nafl, Nef-associated factor 1, that increases cell surface CD4 expression"FEBS Letters. 442(1). 83-88 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Y.Suzuki et al.: "Determinant in human immunodeficiency virus type 1 for efficient replication under cytokine-induced CD4+ T-helper 1(Th1)-and Th2-type conditions"J. Virol.. 73(1). 316-324 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Motoyama et al.: "bcl-x prevents apoptotic cell death of both primitive and definitive erythrocytes at the end of maturation"J. Exp. Med.. 189(11). 1691-1698 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] M.Matsumoto et al.: "Activation of the transcription factor ISGF3 by interferon-gamma"Biol. Chem.. 380(6). 699-703 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] T.Kimura et al.: "Molecular cloning and genomic organization of mouse homologue of Drosophila germ cell-less and its expression in germ lineage cells"Biochem. Biophys. Res. Commun.. 262(1). 223-230 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] A.Kusunoki et al.: "Antisense oligodeoxynucleotide complementary to CXCR4 mRNA block replication of HIV-1 in COS cells"Nucleosides Nucleotides. 18(6-7). 1705-1708 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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