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Functional analysis of oxytocin receptor by generation of oxytocin receptor gene deficient mice.

Research Project

Project/Area Number 09660070
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionTohoku University

Principal Investigator

NISHIMORI Katsuhiko  Tohoku University, Faculty of Agriculture, Department of Applied Biochemistry, Associate Professor, 農学部, 助教授 (10164609)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
KeywordsOxytocin / Receptor / G-protein linked / knockout / conditional / tissues specific
Research Abstract

After the application of this grant, Dr. Kimura (Oosaka-Univ) published the sequence of mouse OTR gene. Based on that sequence, we got OTR cDNA from brain derived cDNA of 1295 vEv mouse. With this cDNA and pGKhprt/MCA1tk vector, we generated OTR-targeting vector. Just after the completion of this vector, we got an important information from my friend in USA, saying that knock-out project for OTR gene, carried out at a Lab. in NIH, USA, gave them unexpected result : the early embryonic death of OTR deficient homozygous mice in utero. With this information, we changed four initial strategy of "conventional knockout of OTR gene" to "conditionalknockout". The conditional knockout are dependent of combination of specificelement loxP and the gene coding bacterial recombinase cre, derived from P1 phage. After establishing of ES cell line, whose targeted gene is homologousely substituted by "fox gene", the ES cells are used to generate the chimeric, and successively the homozygous mutant mice. … More Crossing of floxP mice with the other trasgenic mice, which is harboring exogenous cre gene under control of Tc-inducible promoter, or tissue specific enhancer/promoter, can generate new mutant mice, whose targeted gene is lost and inactivated in specific tissue, or specific stage in embryogenesis. Because of the alteration of experimental situation about OTR gene knockout, I and Dr. Kimura decided to begin this project as a collaboration from April, 1998. We gotlambda phage harboring mouse OTR gene from Dr. KimuRa, and made detailed restriction maps of OTR gene, and generated new OTR knockout vector. This vector comprises neomycin resistant gene, 3 loxP elements, 5'arm and 3'arm of OTR gene, the fragment coding for exon 2 and 3, HSV-tk gene, and vector encoding ori andampicillin resistant gene. The total length of this new vector is 18.5 kb. Now we are introducing this vector to E14TG2a ES cell and screening the cell, whose OTR gene was correctly targeted. In near future, we will use new inducible cre system, in which mutated steroid receptor-cre fusion protein will be activated by the administration of steroid derivative. This mutant mouse system is being developed by Dr. Brown, C.in Baylor College of Medicine, Houston, Texas, USA.For uterus specific OTR geneknock out, we will obtain a mutant mouse which is harboring cre-gene under control of smooth muscle specific actin promoter. This mutant mouse was generated by Dr. Miwa, Osaka university. Less

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Julia A.Elvin et: "Molecular characterization of (the follicle defects in) the growth differentiation factor 9-deficient ovary" Molecular Endocrinology (in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nawa,A.et.al: "Tumor metastasis-associated human MTA1 gene : its sequence analysis and association with cancer cell proliferation." Submitted to Cancer Res.(1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nomura,O et al.: "Determination by modified RT-PCR of transcript amounts from genes involved in sex-steroid synthesis in chicken organs including brain" J.Steroid Biochem.mol.Biol.67. 143-148 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Julia A.Elvin et al.: "Molecular characterization of (the follicle defects in) the growth differentiation factor 9-deficient ovary" MolEndocrinol. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nawa, A.et al.: "Tumor metastasis-associated human MTA1 gene : its sequence analysis and association with cancer cell proliferation" Cancer Res.(to submited).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nomura, O et al.: "Determination by modified RT-PCR of transcript amounts from genes involved in sex-steroid synthesis in chicken organs including brain" J Steroid Biochem Mol Biol. 67. 143-148 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nomura, O: "Determination by modified RT-PCR of transcript omount from gene involved in sex-steroid synthesis in chicken organs including brain" J.Steroid Biochem.Mol.Bicl. 67. 143-148 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Julia A Elvin: "Molecular Characterization of (the follicle delection) the growth differentiation factor-9 deffiueni ovary" Mol Endocrinol. in press. (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Wakabayashi,N.: "The cDNA cloning and transient expression of a chicken gene encoding a follicle-stimulating hormone receptor." Gene. 197. 121-127 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 西森克彦: "生殖に関連したTGF-βスーパーファミリーメンバー" 蛋白質核酸酵素. 43. 491-501 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 西森克彦: "動物の生殖を制御する遺伝子" 化学と生物. 35. 699-708 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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