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Development and its application of simple methods for bone sections

Research Project

Project/Area Number 03557076
Research Category

Grant-in-Aid for Developmental Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Morphological basic dentistry
Research InstitutionShowa University

Principal Investigator

YOSHIKI Shusaku  Showa Univ., Sch.of Dentistry Professor, 歯学部, 教授 (30085740)

Co-Investigator(Kenkyū-buntansha) IKEDA Tohru  Showa Univ., Sch.of Dentistry Assistant Prof., 歯学部, 講師 (00211029)
YAMAGUCHI Akira  Showa Univ., Sch.of Dentistry Associate Prof., 歯学部, 助教授 (00142430)
TACHIKAWA Tetsuhiko  Showa Univ., Sch.of Dentistry Associate Prof., 歯学部, 助教授 (10085772)
KAKUTA Saburou  Showa Univ., Sch.of Dentistry Associate Prof., 歯学部, 助教授 (40112726)
SAKAMOTO Keizou  Showa Univ., Sch.of Medicine Associate Prof., 医学部, 助教授 (50119195)
菱川 健司  昭和大学, 歯学部, 助手 (50189784)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1993: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1991: ¥3,300,000 (Direct Cost: ¥3,300,000)
Keywordsbone tissue / cyanuric chloride / decakcified section / hitochemistry / immunohistochemistry / in situ hybridization / 骨標識 / in situ hybridization
Research Abstract

Dr.Yoshiki, a principal investigator of this research project, reported the simple method to identify osteoid tissues in decalcified bone sections. In this method, treatment of bone tissues with cyanuric chloride before decalcification is essential to identify osteoid tissues. In the present study, we conducted several experiments to develop new application of this technique for bone research, and obtained the following results.
1) When tartrate acid resistant acid phosphatase (TRACP) activity was detected in various tissues prepared by conventional technique, its activity was found is not only osteoclasts but also mononuclear cells in spleen and lungs. However, its activity was detected in only osteoclasts when tissues were prepared after cyanuric chloride treatment. These results indicate that this technique is useful to identify osteoclast cell lineage in decalcified bone sections.
2) We recently developed in situ hybridization technique for bone sections. We examined the effects of cyanuric chloride treatment on in situ hybridization. The expression of mRNAs for osteocalcin and type I collagen is detected in bone sections prepared after cyanuric chloride treatment as well as bone sections prepared by conventional technique. These results suggest that cyanuric chloride treatment dose not affect detection of mRNA by in situ hybridization. This also suggests the possibility of combined technique of histochemistry and in situ hybridization.
3) We tried immunohistochemistry using bone sections treated with cyanuric chloride, but we obtained strong background staining. This technique is not useful for immunohistochemistry.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Nakamura,Y.,et al.: "Acid phosphatase activity is detected preferentially in the osteoclastic lineage by pre-treatment with cyanuric chloride" J.Histochem.Cytochem. 39. 1415-1420 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ikeda,T.,et al.: "In situ hybridization of bone matrix proteins in undecalcified adult rat bone sections" J.Histochem.Cytochem. 40. 1079-1088 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Nakamura, Y., Yamaguchi, A., Ikeda, T, and Yoshiki, S.: "Acid phosphatase activity is detected preferentially in the osteoclastic lineage by pretreatment with cyanuric chloride." J.Histochem.Cytochem. 39. 1415-1420 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ikeda, T., Nomura, S., Yamaguchi, A., Suda, T., and Yoshiki, S.: "In situ hybridization of bone matrix proteins in undecalcified adult rat bone sections." J.Histochem.Cytochem. 40. 1079-1088 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Nakakura,et al.: "Acid phosphatase activity is detected preferentially in the osteoclastic lineage by pre-treatnment with cyanuric chlorid" J.Histochem.Cytochem.39. 1415-1420 (1991)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Ikeda,et al.: "In situ hybridization of bone matrix proteins in undecalcified adult rat bone sections" J.Histochem.Cytochem.40. 1079-1088 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Nakamura et al.: "Acid phosphatase activity is deteted nreferentially in the osteoclastic lineage by pre-treatment with cyanotic chloride" J.Histochem.Cytochem.39. 1415-1420 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Ikeda et al.: "In situ hybridization of bone matrix proteins in undecalcified adult rat bone sections" J.Histochem.Cytochem. 40. 1079-1088 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] C.H.Jin et al.: "1,25-dihydrox-vitamin D3 regulates in vivo production of the third component of complement(C3)in bone" Endocrinol.131. 2468-2475 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Yuich Nakamura: "Acid phosphatase activity is detected preferentially in the osteoclastic lineage by pre-treatment with cyanuric chloride" J.Histochemistry and Cytochemistry. 39. 1415-1420 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Akira Yamaguchi: "Clonal osteogenic cell lines express myogenic and adipocytic developmental potential" Calcified Tissue International. 49. 221-225 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Akira Yamaguchi: "Recombinant human bone morphogenetic protein-2 stimulates osteoblastic maturation and inhibits myogenic differentiation in vitro" J.Cell Biology. 111. 681-687 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Tohru Ikeda: "cDNA and deduced amino acid sequence of mouse matrix gla protein" J.Bone and Mineral Research. 6. 1013-1017 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yoshiko Ishimi: "IL-6 is produced by osteoblasts and induces bone resorption" J.Immunology. 145. 3297-3303 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Takuhiko Akatsu: "Role of prostaglandins in Interleukin-1-induced bone resorption in mice in vitro" J.Bone and Mineral Research. 6. 183-190 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 山口 朗: "骨・歯牙組織の病理検査法と研究技術の実際" 学際企画, 12 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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