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Functional analysis of germ line cells using gene manipulated mouse

Research Project

Project/Area Number 11234203
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionOSAKA UNIVERSITY

Principal Investigator

OKABE Masaru  Osaka University, Genome Information Research Center, Professor, 遺伝子情報実験センター, 教授 (30089875)

Co-Investigator(Kenkyū-buntansha) KONDOH Gen  Osaka University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (40243258)
Project Period (FY) 1999 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥85,000,000 (Direct Cost: ¥85,000,000)
Fiscal Year 2002: ¥20,900,000 (Direct Cost: ¥20,900,000)
Fiscal Year 2001: ¥20,900,000 (Direct Cost: ¥20,900,000)
Fiscal Year 2000: ¥20,900,000 (Direct Cost: ¥20,900,000)
Fiscal Year 1999: ¥22,300,000 (Direct Cost: ¥22,300,000)
Keywordsgene manipulate animals / transgenic mouse / GFP / sex differentiation / sex chimera / ACE / GPI-anchor / 生殖細胞 / 複数分裂 / 雌雄キメラ / Cre / 1oxP / loxP / 雌雄選別 / キメラ
Research Abstract

We have developed an easy and very efficient system to separate male and female embryos before implantation stage with non invasive manner. When male and female eggs prepared by this system were aggregated to produce sex chimera, the advantage of this system is the easiness of pointing out the sex of individual cells that form organs by observing the transgenically incorporated green fluorescent protein gene in X chromosome. By analyzing the genital organs in thus prepared sex chimeras, we have found many spermatogonia like cells of female origin in phenotypically male testes. The pattern of the genomic imprinting of these genes was found out to be male type in spite of their original sex was female.
With the collaboration to Prof. Nakatsuji's group, it was also found out that these "female spermatogonia" cells entered meiosis in early stages of development However, on the other hand, some cells originated from female became huge cells like eggs in the testis of sex chimera animals, whi … More le the majority of the female cells developed into male type spermatogonia. The complexity of the sex differentiation of germ cells was elucidated by producing these sex chimera mice and simultaneously provided a tool to analyze the mechanism of male-female differentiation in germ line cells.
Pig-a, an X-linked gene, is a key component of glycosylphosphatidylinositol (GPI) anchor biosynthesis based on the fact that lack of this gene causes deficiencies of hundreds of GPI-anchored proteins. Dr. Kondoh has produced a transgenic mouse line that expressed enhanced green fluorescent protein in GPI-anchored form and monitored GPI-anchor-positive cells in situ. It was found out that ACE has phospholipase activity other than the dipeptidyl-peptidase. It was suggested that testicular ACE is indispensable because it may release GPI anchored proteins from sperm surface to make fertile sperm. The results indicated that the necessity of re-evaluation of the involvement of ACE in reproductive biology. Less

Report

(5 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Inoue, N. et al.: "Disruption of Mouse CD46 Causes an Accelerated Spontaneous Acrosome Reaction in Sperm"Mol Cell Biol. 23. 2614-2622 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Baba, D. et al.: "Mouse Sperm Lacking Cell Surface Hyaluronidase PH-20 Can Pass through the Layer of Cumulus Cells and Fertilize the Egg"J Biol Chem. 277. 30310-30314 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Gao, X.H.et al.: "Rapid compensation for glycosylphosphatidylinositol anchor deficient keratinocytes after birth: visualization of glycosylphosphatidylinositol-anchored proteins in situ"J Invest Dermatol. 118. 998-1002 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hasuwa, H. et al.: "Small interfering RNA and gene silencing in transgenic mice and rats"FEBS Left et al.. 532. 227-230 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakanishi, T. et al.: "FISH Analysis of 142 EGFP Transgene Integration Sites into the Mouse Genome"Genomics. 80. 564-574 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yamagata, K. et al.: "Sperm from the calmegin-deficient mouse have normal abilities for binding and fusion to the egg plasma membrane"Dev Biol. 250. 348-357 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 蓮輪 英毅ほか: "RNAi実験プロトコール"羊土社. 178 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inoue, N. et al.: "Disruption of Mouse CD46 Causes an Accelerated Spontaneous Acrosome Reaction in Sperm"Mol Cell Biol. 23(7). 2614-2622 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Baba, D. et al.: "Mouse Sperm Lacking Cell Surface Hyaluronidase PH-20 Can Pass through the Layer of Cumulus Cells and Fertilize the Egg"J Biol Chem. 277(33). 30310-30314 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Gao, X. H. et al.: "Rapid compensation for glycosylphosphatidylinositol anchor deficient keratinocytes after birth : visualization of glycosylphosphatidylinositol-anchored proteins in situ"J Invest Dermatol. 118(6). 998-1002 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hasuwa, H. et al.: "Small interfering RNA and gene silencing in transgenic mice and rats"FEBS Lett. 532(1-2). 227-230 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakanishi, T., et al.: "FISH Analysis of 142 EGFP Transgene Integration Sites into the Mouse Genome"Genomics. 80(6). 564-574 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yamagata, K. et al.: "Sperm from the calmegin-deficient mouse have normal abilities for binding and fusion to the egg plasma membrane"Dev Biol.. 250(2). 348-357 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hasuwa, H. et al.: "Yodosha"RNAi experiment protocol. 178 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kimura T et al.: "Mouse germ cell-less as an essential component for nuclear integrity"Mol Cell Biol. 23(4). 1304-1315 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Baba D et al.: "Mouse sperm lacking cell surface hyaluronidase PH-20 can pass through the layer of cumulus cells and fertilize the egg"J Biol Chem. 277(33). 30310-30314 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Murakami H et al.: "Transgenic pigs expressing human decay-accelerating factor regulated by porcine MCP gene promoter"Mol Reprod Dev. 61(3). 302-311 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yamagata K et al.: "Sperm from the calmegin-deficient mouse have normal abilities for binding and fusion to the egg plasma membrane"Dev Biol. 250(2). 348-357 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nomura M et al.: "Mechanism of host cell prptection from complement in murine cytomegalovirus (CMV) infection : identification of a CMV-responsive element in the CD46 promoter region"Eur J Immunol. 32(10). 2954-2964 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakanishi T et al.: "FISH Analysis of 142 EGFP Transgene Integration Sites into the Mouse Genome"Genomics. 80(6). 564.574 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakanishi T. et al.: "Alkalinization of Acrosome Measured by GFP as a pH Indicator and Its Relation to Sperm Capacitation"Dev.Biol.. 237. 222-231 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Ikawa M. et al.: "Calmegin Is Required for Fertilin α/β Heterodimerization and Sperm Fertility"Dev.Biol.. 240. 254-261 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Funahashi H. et al.: "Nuclear Transfer of Blastomeres Expressing EGFP-Reporter Gene May Improve the Efficiency of Transgenic Cattle"CLONING AND STEM CELLS. 3. 183-190 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Horie K. et al.: "Efficient chromosomal transposition of a Tc1/mariner-like transposon Sleeping Beauty in mice"Proc.Natl.Acad.Sci.. 98. 9191-9196 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kimura SH. et al.: "Cyclin G1 is involved in G2/M arrest in response to DNA damage and in growth control after damage recovery"Oncogene. 20. 3290-3300 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kawane K. et al.: "Requirement of DNase II for defenitive erythropoiesis in the mouse fetal liver"Science. 292. 1546-1549 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 中西 友子 他: "体外受精-Update"鈴木 秋悦. 360 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Zhou,L. et al.: "Two independent pathways of maternal cell transmission to offspring : through placenta during pregnancy and by breast-feeding after birth"Immunology. 101(4). 570-80 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Murakami,H. et al.: "Production of transgenic pigs expressing human DAF (CD55) regulated by the porcine MCP gene promoter"Transplant Proc. 32(7). 2505-6 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Miyado,K. et al.: "Requirement of CD9 on the egg plasma membrane for fertilization."Science. 287(5451). 321-4 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Koresawa,Y. et al.: "A new Cre recombinase gene based on optimal codon usage in mammals : a powerful material for organ-specific gene targeting"Transplant Proc. 32(7). 2516-7 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Koresawa,Y. et al.: "Synthesis of a New Cre Recombinase Gene Based on Optimal Codon Usage for Mammalian Systems"J Biochem (Tokyo). 127(3). 367-72 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ando,H. et al.: "Spermatocyte-specific gene excision by targeted expression of Cre recombinase"Biochem Biophys Res Commun. 272(1). 125-8 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kondoh G. et al.: "Tissue-inherent fate of GPI revealed by GPI-anchored GFP transgenesis"FEBS Lett. 458. 299-303 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nakanishi T. et al.: "Real-time observation of acrosomal dispersal from mouse sperm using GFP as a marker protein"FEBS Lett. 449(2-3). 277-283 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2018-03-28  

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