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An immuno-electron microscopic study of the localization of cathepsins B and L and collagenase in cultured human odontoclasts

Research Project

Project/Area Number 07671975
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphological basic dentistry
Research InstitutionKyushu University

Principal Investigator

TANAKA Teruo  Kyushu University, Faculty of Dentistry, Professor, 歯学部, 教授 (60077667)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsOdontoclast / Osteoclast / Cathepsin L / Collagen / Cystatin / Immunoelectron microscopy / 免疫電頭 / 骨 / カテプシンB / コラゲナーゼ / ヒト / 微細構造 / 歯根吸収
Research Abstract

We isolated human odontoclasts from primary teeth and cultured them on the culture dishes or dentine slices with alpha-MEM and 10% FCS.Human odontoclasts were tertrate-resistant acid phosphatase positive and formed resorption lacunae on the dentine slices. We examined the effect ot 5 mU/ml calcitonin on the morphological changes of odontoclasts, but no morphological changes were observed. Because of the difficulty of the isolation of odontoclasts and lack of the supply of the samples (primary teeth), we could not obtain many data from cultured human odontoclasts. Further investigations are needed for finding the efficient-isolation techniques and an optimal condition for human odontoclasts.
We also examined the changes in the immunocytochemical localization of cathepsin L,type I collagen and rat salivary cystatin-3 (RSC-3) in rat osteoclasts treated with E-64. The immunoreactivity for cathepsin L was extracellularly very weak, but was strong intracellurlarly in the E-64-treated osteoclasts. The intracellular immuno-reactivity for type I collagen was found inthe vacuoles in the E-64-treated osteoclasts but not in any vacuoles in the control osteoclasts. In the immunoreaction for RSC-3 in the E-64-treated, the cytoplasm of the ruffled border was positive, and also the tips of the RSC-3 positive-ruffled border appeared to deeply enter into the bone matrix. Intracellularly, the granular reaction products of RSC-3 were found in vacuoles. These finding suggests that in the E-64-treated osteoclasts, the extracellular release of cathepsin L was insufficient to suppress the degradation of collagen, and that RSC-3 is probably associated wit the inhibition of cathepsin L in the lysosomes in the osteoclasts.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Ryouji Moroi: "The changes in the Immunocyochemical Localization of Cathepsin L and Type Collagen in Rat Osteoclast Treated with E-64" Acta Histochemica et Cytochemica. 28.6. 523-531

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] R.Moroi: "An immunocytochemical study of cathepsin L and rat salivary cystatin-3 in rat osteoclast treated E-64." Archives of Oral Biology. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Moroi R,Yamaza T,Ayukawa Y,Kiyoshima T,Cytochem Ohsaki Y,Nishimura Y,Terada Y,Himeno M,and Tanaka T: "The changes in the immunocytochemical localization cathepsin L and type I collagen in Rat osteoclasts treated with E-64" Acta Histochem.Vol.28 No.6. (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Moroi R,Yamaza T,Nishimura Y,Terada Y: "An immunocytochemiccal study of cthepsin L and rat salivary cystatin-3 in rat osteoclasts treated with E-64." Archives of Oral Biology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ryouji Moroi: "The changes in the lmmunocyochemical Localization of Cathepsin L and Typel Collagen in Rat Osteoclast Treated with E-64" Acta Hitochemica et Cytochemica. 28.6. 523-531 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] R.Moroi: "An immunocytochemical study of cathepsin L and rat salivary cystatin-3 in rat osteoclast treated E-64." Archives of Oral Biology. (印刷中).

    • Related Report
      1996 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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