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Localization of endogenous osteocalcin in neonatal rat bone and its absence in articular cartilage: Effect of warfarin treatment

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Summary

Immunocytochemistry after cryoultramicrotomy was used to localize endogenous osteocalcin in bone (calvaria, femoral diaphysis) and epiphyseal femoral cartilage from 8-day-old rats treated (or mot) for 7 days with warfarin. Ultrathin frozen sections were incubated with goat antiserum against rat osteocalcin at high dilutions (2×10−4 to 2×10−6). In calvaria and femur of untreated rats, endogenous osteocalcin was observed in osteoblasts (cytoplasm and nucleus) and in the collagenous matrix. Osteocalcin appeared progressively in osteoblasts and bone matrix in the mineralization front, then increased in the regions of extended calcification. Osteocalcin was also detected in osteocytes but was not as abundant as in osteoblasts. In bone samples of warfarin-treated rats, endogenous osteocalcin was only detected in bone matrix but not in osteoblasts. Furthermore, osteocalcin was only observed if antiserum was not very dilute (2×10−2). In cartilage (hypertrophied and degenerative zones), osteocalcin was not observed in matrix and chondrocytes. However, it was found in the vicinity of matrix vesicles at the initial loci of calcification. Osteocalcin was never detected in the cartilage of warfarin-treated rats. Our results provide ultrastructural immunocytological evidence for the localization of endogenous osteocalcin in osteoblasts, the presence of osteocalcin in bone matrix and a direct gradient between the presence of osteocalcin and the calcification process. Osteocalcin is absent from cartilage, except possibly close to calcifying matrix vesicles. Warfarin inhibits the formation of osteocalcin.

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Authors and Affiliations

  1. Faculté A. Carrel, INSERM Unité 234, rue G. Paradin, F-69372, Lyon Cedex 08, France

    Georges Boivin, Chantal Anthoine-Terrier & Pierre J. Meunier

  2. Faculté Lyon-Sud, CNRS URA 559, Oullins, France

    Gérard Morel & Paul M. Dubois

  3. Laboratory for the Study of Skeletal Disorders, Harvard Medical School, Boston, USA

    Jane B. Lian

  4. Department of Cell Biology, University of Massachusetts Medical Center, Worcester, Massachusetts, USA

    Jane B. Lian

Authors
  1. Georges Boivin
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  2. Gérard Morel
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  3. Jane B. Lian
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  4. Chantal Anthoine-Terrier
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  5. Paul M. Dubois
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  6. Pierre J. Meunier
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Boivin, G., Morel, G., Lian, J.B. et al. Localization of endogenous osteocalcin in neonatal rat bone and its absence in articular cartilage: Effect of warfarin treatment. Vichows Archiv A Pathol Anat 417, 505–512 (1990). https://doi.org/10.1007/BF01625731

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  • DOI: https://doi.org/10.1007/BF01625731

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