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Studies on the Regulation of Fertilization and Activation of Development

Research Project

Project/Area Number 12306018
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied molecular and cellular biology
Research InstitutionUniversity of Tsukuba

Principal Investigator

BABA Tadashi  University of Tsukuba, Institute of Applied Biochemistry, Professor, 応用生物化学系, 教授 (40165056)

Co-Investigator(Kenkyū-buntansha) NISHIMURA Hitoshi  University of Tsukuba, Institute of Applied Biochemistry, Assistant Professor, 応用生物化学系, 講師 (80241347)
OKABE Masaru  Osaka University, Genome Information Research Center, Professor, 遺伝情報実験センター, 教授 (30089875)
OKAMURA Naomichi  University of Tsukuba, Institute of Applied Biochemistry, Professor, 基礎医学系, 教授 (30134224)
NAKANISHI Tomoko  University of Tsukuba, Institute of Applied Biochemistry, Assistant Professor, 応用生物化学系, 助手 (10344863)
OGURA Atsuo  The Institute of Physical and Chemical Research (RIKEN), Bioresource Center, Head of Bioresource Engineering Division, 遺伝工学基盤技術室, 室長 (20194524)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥44,120,000 (Direct Cost: ¥37,100,000、Indirect Cost: ¥7,020,000)
Fiscal Year 2002: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
Fiscal Year 2001: ¥18,720,000 (Direct Cost: ¥14,400,000、Indirect Cost: ¥4,320,000)
Fiscal Year 2000: ¥13,700,000 (Direct Cost: ¥13,700,000)
KeywordsSperm / Egg / Fertilization / Sperm maturation / Egg activation / Cell fusion / Oviduct / Knockout mouse / 卵活性化
Research Abstract

To elucidate the regulation of fertilization and activation of development, we have examined possible functions of mouse sperm in sperm maturation, sperm transit from uterus to oviduct, fertilization, and egg activation. The following experimental results have been obtained :
1. The TritonX-100-insoluble microdomains, lipid rafts, on the plasma membranes of epididymal sperm gradually move to the apical region from the equatorial region of sperm during capacitation.
2. Male mice lacking sperm PH-20 are still fertile, in spite of a reduced ability to disperse cuwnulus cells from the cumulus mass. Western blot analysis of sperm extracts reveals the presence of other hyaluronidase(s), except PH-20, presumably within the sperm acrosome. These data provide evidence that PH-20 is not essential for fertilization at least in the mouse, and suggest that the other hyaluronidase(s) may play an important role in the sperm penetration through the cumulus cell layer and/or the egg zona pellucida, possi … More bly in cooperation with PH-20.
3. Mouse genomic clones encoding a 42-kDa sperm serine protease, TESP5, have been identified. Immunochemical analysis reveals that 42- and 4l-kDa forms of TESP5 are localized in the head, cytoplasmic droplet, and tail of cauda epididymal sperm probably as a glycosylphosphatidylinositol-anchored membranous protein. Moreover, these two forms of TESP5 are selectively included into Triton X-100-insoluble microdomains, Iipid rafts, of the sperm membranes. These results suggest a possible involvement of TESP5 in the sperm/egg interactions.
4. Two different ADAMI isoforms, ADAMla and ADAMlb, instead of ADAMl are present in the mouse testis. ADAMla is resttrictedly present within the endoplasmic reticulum of germ cells, whereas epididymal sperm contain only ADAMlb on the cell surface. The precursors of ADAMla and ADAMlb form a heterodimeric complex with that of ADAM2 in the endoplasmic reticulum of germ cells. The heterodimeric complex between the mature forms of ADAMlb and ADAM2 is also found on the sperm surface. These data imply the potential roles of ADAMla and ADAMlb in spermatogenesis and fertilization, respectively.
5. When a synthesized mRNA with a poly(A) tail encoding phospholipase ζ is injected into unfertilized eggs, pronuclear formation is observed. Less

Report

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

    (44 results)

All Other

All Publications (44 results)

  • [Publications] Atsuo Inoue: "The transcript for a novel protein with a zinc finger motif is expressed at specific stages of mouse spermatogenesis"Biochem. Biophys. Res. Commun.. 273. 398-403 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tsuneatsu Mori: "Molecular and immunological approaches to mammalian fertilization"J. Reprod. Immunol.. 47. 139-158 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shin-ichi Kashiwabara: "Identification of a novel isoform of poly(A) polymerase, TPAP, specifically present in the cytoplasm of spermatogenic cells"Dev. Biol.. 228. 106-115 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Eri Kodama: "CDNA cloning and functional analysis of ascidian sperm proacrosin"J. Biol. Chem.. 276. 24594-24600 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hitoshi Nishimura: "The ADAM1a and ADAM1b genes, instead of the ADAM1 (fertilin α) gene, are localized on mouse chromosome 5"Gene. 291. 67-76 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Eri Kodama: "Spermosin, a trypsin-like protease from ascidian sperm : cDNA cloning, protein structures and functional analysis"Eur. J. Biochem.. 269. 657-663 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Arata Honda: "A mouse serine protease TESP5 is selectively included into lipid rafts of sperm membrane presumably as a glycosylphosphatidylinositol-anchored protein"J. Biol. Chem.. 277. 16976-16984 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Jindawan Siruntawineti: "Occurrence of small, round vesicles in the acrosome of elongating spermatids from a mouse mutant line with a partial deletion of the Y chromosome"J. Reprod. Dev.. 48. 513-521 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Arata Honda: "Role of acrosomal matrix proteases in sperm-zona pellucida interactions"Hum. Reprod. Update. 8. 405-412 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Daichi Baba: "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] Shin-ichi Kashiwabara: "Regulation of spermatogenesis by testis-specific, cytoplasmic poly(A) polymerase TPAP"Science. 298. 1999-2002 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shin Sugiura: "Expression of a testis-specific form of TBP-related factor 2 (TRF2) mRNA during mouse spermatogenesis"J. Reprod. Dev.. 49. 107-111 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ekyune Kim: "Differential localization of ADAM1a and ADAM1b in the endoplasmic reticulum of testicular germ cells and on the surface of epididymal sperm"Biochem. Biophys. Res. Commun.. 304. 313-319 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inoue, A., et al.: "The transcript for a novel protein with a zinc finger motif is expressed at specific stages of mouse spermatogenesis."Biochem. Biophys. Res. Commun.. 273. 398-403 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Mori, T., et al.: "Molecular and immunological approaches to mammalian fertilization"J. Reprod. Immunol.. 47. 139-158 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kashiwabara, S., et al.: "Identification of a novel isoform of poly(A) polymerase, TPAP, specifically present in the cytoplasm of spermatogenic cells"Dev. Biol.. 228. 106-115 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kodama, E., et al.: "cDNA cloning and functional analysis of ascidian sperm proacrosin"J. Biol. Chem.. 276. 24594-24600 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nishimura. H., et al.: "The ADAMla and ADAMlb genes, instead of the ADAM1 (fertilin α) gene, are localized on mouse chromosome 5"Gene.. 291. 67-76 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kodama, E., et al.: "Spermosin, a trypsin-like protease from ascidian sperm : cDNA cloning, protein structures and functional analysis"Eur. J. Biochem.. 269. 657-663 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Honda, A., et al.: "A mouse serine protease TESP5 is selectively included into lipid rafts of sperm membrane presumably as a glycosylphosphatidylinositol-anchored protein"J. Biol. Chem.. 277. 16976-16984 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Siruntawineti, J., et al.: "Occurrence of small, round vesicles in the acrosome of elongating spermatids from a mouse mutant line with a partial deletion of the Y chromosome"J. Reprod. Dev.. 48. 513-521 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Honda, A., et al.: "Role of acrosomal matrix proteases in sperm-zona pellucida interactions"Hum. Reprod. Update. 8. 405-412 (2002)

    • 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. 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] Kashiwabara, S., et al.: "Regulation of spermatogenesis by testis-specific, cytoplasmic poly(A) polymerase TPAP"Science. 298. 1999-2002

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sugiura, S., et al.: "Expression of a testis-specific form of TBP-related factor 2 (TRF2) mRNA during mouse spermatogenesis"J. Reprod. Dev.. 49. 107-111 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kim, E., et al.: "Differential localization of ADAMla and ADAMlb in the endoplasmic reticulum of testicular germ cells and on the surface of epididymal sperm"Biochem. Biophys. Res. Commun.. 304. 313-319 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hitoshi Nishimura: "The ADAM1a and ADAM1b genes, instead of the ADAM1(fertilin α) gene, are localized on mouse choromosome 5"Gene. 291. 67-76 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Araa Honda: "A mouse serine protease TESP5 is selectively included into lipid rafts of sperm membrane presumably as a glycosylphosphatidylinositol-anchored protein"Journal of Biological Chemistry. 277. 16976-16984 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Jindawan Siruntawineti: "Occurrence of small, round vesicles in the acrosome of elongating spermatids from a mouse mutant line with a partial deletion of the Y chromosome"Journal Reproduction and Development. 48. 513-521 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Daichi Baba: "Mouse sperm lacking cell surface hyaluronidase PH-20 can pass through the layer of cumulus cells and fertilize the egg"Journal of biological Chemistry. 277. 30310-30314 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shin-ichi Kashiwabara: "Regulation of spermatogenesis by testis-specific, cytoplasmic poly(A) plymerase TPAP"Science. 298. 1999-2002 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shin Sugiura: "Expression of a testis-specific form of TBP-related factor 2 (TRF2) mRNA during mouse spermatogenesis"Journal Reproduction and Development. 49. 107-111 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Arata Honda: "A mouse serine protease TESP5 is selectively included into lipid rafts of sperm membrane presumably as a GPI-anchored protein."Journal of Biological Chemistry. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Shin-ichi Kashiwabara: "Role of acrosomal matrix proteases in sperm-zona pellucida interactions."Human Reproduction Update. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Eri Kodama: "Spermosin,a trypsin-like protease from ascidian sperm:cDNA cloning,protein structures and functional analysis."European Journal of Biochemistry. 269. 657-663 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hitoshi Nishimura: "The ADAM1a and ADAM1b genes,instead of the ADAM1(fertilin α)gene,are localized on mouse chromosome 5."Gene. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Eri Kodama: "cDNA cloning and functional analysis of ascidian sperm proacrosin."Journal of Biological Chemistry. 276. 24594-24600 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tomoko Nakanishi: "Alkalinization of acrosome measured by GFP as a pH indicator and its relation to sperm capacitation."Developmental Biology. 237. 222-231 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tsuneatsu Mori: "Molecular and immunological approaches to mammalian fertilization"Journal of Reproductive Immunology. 47. 139-158 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Atsuto Inoue: "The transcript for a novel protein with a zinc finger motif is expressed at specific stages of mouse spermatogenesis"Biochemical and Biophysical Research Communications. 273. 398-403 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Shin-ichi Kashiwabara: "Identification of a novel isoform of poly (A) polymerase, TPAP, specifically present in the cytoplasm of spermatogenic cells"Developmental Biology. 228. 106-115 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kenji Miyado: "Requirement of CD9 on the egg plasma membrane for fertilization"Science. 287. 321-324 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yukie Koresawa: "Synthesis of a new Cre recombinase gene based on optimal codon usage for mammalian systems"Journal of Biochemistry. 127. 367-372 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hitoshi Nishimura: "Analysis of loss of adhesive function in sperm lacking cyritestin and fertilin β"Developmental Biology. (In press). (2001)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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