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Characterization of steroid membrane receptor in fish oocyte and its role in the induction of oocyte maturation

Research Project

Project/Area Number 17570175
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Developmental biology
Research InstitutionShizuoka University

Principal Investigator

TOKUMOTO Toshinobu  Shizuoka University, Faculty of Science, Associate Professor, 理学部, 助教授 (30273163)

Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥2,600,000 (Direct Cost: ¥2,600,000)
Keywordsmembrane progestin receptor alpha / mPRα / oocyte maturation / diethylstilbestrol / DES / endocrine disrupting chemicals / goldfish / プロゲスチン膜受容体 / mPR / 内分泌かく乱物質
Research Abstract

Oocyte maturation in lower vertebrates is triggered by maturation-inducing steroid (MIS), which acts on receptors located on the oocyte membrane and induces the activation of maturation-promoting factor in the oocyte cytoplasm. During the course of maturation, oocytes undergo drastic morphological changes associated with progression of the meotic cell cycle, among which breakdown of the oocyte nuclear envelope (germinal vesicle breakdown, GVBD) occuring at the prophase/metaphase transition is usually regarded as a hallmark of the progress of oocyte maturation. We conducted the cloning of the membrane progestin receptor (mPR) cDNAs from a goldfish ovarian cDNA library. Four clones for mPR subtypes were obtained by RACE-PCR from goldfish ovary. Among these further characterization was performed on mPRα. Northern blot analysis indicates the presence of a major 2.6 kb transcript in ovaries that encodes a 354 amino acid protein which shows high sequence identity with seatrout (81%), zebrafi … More sh (93%) and human (55%) mPRαs. Western blot analysis using a polyclonal goldfish mPRα antibody shows a major immunoreactive band of the predicted molecular weight (40 kDa) in goldfish ovarian membranes. Computer modeling predicts that the deduced protein has seven transmembrane domains, typical of G protein-coupled receptors. Treatment of full grown, late vitellogenic stage follicle-enclosed oocytes in vitro with gonadotropin increased mPRα protein levels. A correlation between mPRα protein levels and the ability of oocytes to undergo GVBD in response to the MIS (maturational competence) was observed after treatment with gonadotropin. Microinjection of goldfish oocytes with a morpholino antisense oligonucleotide to mPRα blocked both the induction of oocyte maturational competence and mPRα protein upregulation by gonadotropin. These results with the goldfish mPRα protein are similar to those obtained previously with spotted seatrout, further supporting the hypothesis that the mPRα acts as an intermediary in MIS induction of oocyte maturation in teleosts.
Further evidence was obtained by using active recombinant protein expressed in human cultured cells. It was demonstrated that both the ovarian plasma membrane receptor in goldfish and goldfish mPRα expressed in transfected cells displayed high affinity, limited capacity, displaceable specific binding for [^3H]-17,20β-DHP. The relative binding affinities of various steroids and EDCs to these two receptor preparations were similar, supporting the idea that mPRα is a MIS receptor in vivo Less

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (28 results)

All 2007 2006 2005

All Journal Article (25 results) Book (2 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Degradation of nucleoproteins by ubiquitin-proteasome system in rat spermatids and human sperm2007

    • Author(s)
      Celina Haraguchi
    • Journal Title

      Journal of Histochemistry and Cytochemistry (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Interactions of diethylstilbestrol (DES) and DES analogues with membrane progestin receptor α (mPRα) and the correlation with their nongenomic progestin activities2007

    • Author(s)
      Toshinobu Tokumoto
    • Journal Title

      Endocrinology (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Degradation of nucleoproteins by ubiquitin-proteasome system in rat spermatids and human sperm.2007

    • Author(s)
      Celina Haraguchi
    • Journal Title

      Journal of Histochemistry and Cytochemistry (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Interactions of diethylstilbestrol (DES) and DES analogues with membrane progestin receptor a (mPRα) and the correlation with their nongenomic progestin activities.2007

    • Author(s)
      Toshinobu Tokumoto
    • Journal Title

      Endocrinology (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Degradation of nucleoproteins by ubiquitin-proteasome system in rat spermatids and human sperm2007

    • Author(s)
      Celina Haraguchi
    • Journal Title

      Journal of Histochemistry and Cytochemistry (In press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Interactions of diethylstilbestrol (DES) and DES analogues with membrane progestin receptor α(mPRα) and the correlation with their nongenomic progestin activities2007

    • Author(s)
      Toshinobu Tokumoto
    • Journal Title

      Endocrinology (In press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Cloning and identification of a membrane progestin receptor in goldfish ovaries and evidence it is an intermediary in oocyte meiotic maturation2006

    • Author(s)
      Mika Tokumoto
    • Journal Title

      General and Comparative Endocrinology 145・1

      Pages: 101-108

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Comparative proteome analysis of changes in the 26S proteasome during oocyte maturation in goldfish2006

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Proteomics 6・14

      Pages: 4195-4202

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Cytoplasmic lipid droplets are sites of convergence of proteasomal and autophagic degradation of apolipoprotein B2006

    • Author(s)
      Yuki Ohsaki
    • Journal Title

      Molecular Biology of the Cell 27・1

      Pages: 2674-2683

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Molecular Cloning of cDNA Encoding APC11, a Catalytic Component of Anaphase-promoting-complex (APC/C), from Goldfish (Carassius auratus) and Establishment of in vitro Ubiquitinating System2006

    • Author(s)
      Mika Tokumoto
    • Journal Title

      Zoological Science 23・2

      Pages: 675-678

    • NAID

      110006151749

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Cloning and identification of a membrane progestin receptor in goldfish ovaries and evidence it is an intermediary in oocyte meiotic maturation.2006

    • Author(s)
      Mika Tokumoto
    • Journal Title

      General and Comparative Endocrinology 145-1

      Pages: 101-108

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Comparative proteome analysis of changes in the 26S proteasome during oocyte maturation in goldfish.2006

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Proteomics 6-14

      Pages: 4195-4202

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Cytoplasmic lipid droplets are sites of convergence of proteasomal and autophagic degradation of apolipoprotein B.2006

    • Author(s)
      Yuki Ohsaki
    • Journal Title

      Molecular Biology of the Cell 27-1

      Pages: 2674-2683

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Molecular Cloning of cDNA Encoding APC 11, a Catalytic Component of Anaphase-promoting-complex (APC/C), from Goldfish (Carassius auratus) and Establishment of in vitro Ubiquitinating System.2006

    • Author(s)
      Mika Tokumoto
    • Journal Title

      Zoological Science 23-2

      Pages: 675-678

    • NAID

      110006151749

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Molecular cloning of cDNA encoding APC11, a catalytic component of anaphase-promoting-complex (APC/C), from goldfish (Carassius auratus) and establishment of in vitro ubiquitinating system2006

    • Author(s)
      Mika Tokumoto
    • Journal Title

      Zoological Science 23・8

      Pages: 675-678

    • NAID

      110006151749

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Cloning and identification of a membrane progestin receptor in goldfish ovaries and evidence it is an intermediary in oocytemeiotic maturation2006

    • Author(s)
      Mika Tokumoto
    • Journal Title

      General and Comparative Endocrinology 145・1

      Pages: 101-108

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification of a protein kinase which phosphorylates a subunit of the 26S proteasome and changes in its activity during meiotic cell cycle in goldfish oocytes.2005

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Cellular Signalling 17・2

      Pages: 205-215

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Molecular cloning and expression of cDNA coding for four spliced isoforms of Casein kinase Iα in goldfish oocytes2005

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Biochimica et Biophysica Acta 1727・1

      Pages: 75-80

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report
  • [Journal Article] The cytoskeleton-dependent localization of cdc2/cyclin B in blastomere cortex during Xenopus embryonic cell cycle2005

    • Author(s)
      Norihiko Nakamura
    • Journal Title

      Molecular Reproduction and Development 72・3

      Pages: 336-345

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report
  • [Journal Article] Induction and inhibition of oocyte maturation by EDCs in zebrafish2005

    • Author(s)
      Toshinobu Tokumoto
    • Journal Title

      Reproductive Biology and Endocrinology 3・69

      Pages: 1-9

    • NAID

      120000796499

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report
  • [Journal Article] Identification of a protein kinase which phosphorylates a subunit of the 26S proteasome and changes in its activity during meiotic cell cycle in goldfish oocytes.2005

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Cellular Signalling 17-2

      Pages: 205-215

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Molecular cloning and expression of cDNA coding for four spliced isoforms of Casein kinase Iα in goldfish oocytes.2005

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Biochimica et Biophysica Acta 1727-1

      Pages: 75-80

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The cytoskeleton-dependent localization of cdc2/cyclin B in blastomere cortex during Xenopus embryonic cell cycle.2005

    • Author(s)
      Norihiko Nakamura
    • Journal Title

      Molecular Reproduction and Development 72-3

      Pages: 336-345

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Induction and inhibition of oocyte maturation by EDCs in zebrafish.2005

    • Author(s)
      Toshinobu Tokumoto
    • Journal Title

      Reproductive Biology and Endocrinology 3-69

      Pages: 1-9

    • NAID

      120000796499

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Identification of a protein kinase which phosphorylates a subunit of the 26S proteasome and changes in its activity during meiotic cell cycle in goldfish oocytes2005

    • Author(s)
      Ryo Horiguchi
    • Journal Title

      Cellular Signalling 17・2

      Pages: 205-215

    • Related Report
      2005 Annual Research Report
  • [Book] New impact on protein modifications in the regulation of reproductive system2007

    • Author(s)
      Toshinobu Tokumoto 編
    • Total Pages
      167
    • Publisher
      Research Signpost
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] New impact on protein modifications in the regulation of reproductive system2005

    • Author(s)
      Toshinobu Tokumoto編
    • Total Pages
      167
    • Publisher
      Research Signpost
    • Related Report
      2006 Annual Research Report
  • [Patent(Industrial Property Rights)] 内分泌撹乱性物質のスクリーニング方法2005

    • Inventor(s)
      徳元俊伸
    • Industrial Property Rights Holder
      静岡大学
    • Industrial Property Number
      2005-267894
    • Filing Date
      2005-09-15
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary

URL: 

Published: 2005-04-01   Modified: 2016-04-21  

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