Journal of Bone and Mineral Metabolism

, Volume 21, Issue 6, pp 344–352

Suppression of differentiation and proliferation of early chondrogenic cells by Notch

Authors

  • Naoko Watanabe
    • Research Institute for Biological SciencesTokyo University of Science
  • Yoko Tezuka
    • Department of Tissue and Organ Development, Regeneration and Advanced Medical ScienceGifu University Graduate School of Medicine
  • Kenji Matsuno
    • Department of Biological Science and TechnologyTokyo University of Science
  • Seiji Miyatani
    • The Institute of Physical and Chemical Research (RIKEN)
  • Naoko Morimura
    • Research Institute for Biological SciencesTokyo University of Science
  • Masafumi Yasuda
    • Research Institute for Biological SciencesTokyo University of Science
  • Ryoji Fujimaki
    • Department of Orthopaedic Surgery, School of MedicineKeio University
  • Kazuki Kuroda
    • Department of Medical Chemistry, Faculty of MedicineKyoto University
  • Yuji Hiraki
    • Institute for Frontier Medical SciencesKyoto University
  • Nobumichi Hozumi
    • Research Institute for Biological SciencesTokyo University of Science
  • Ken-ichi Tezuka
    • Department of Tissue and Organ Development, Regeneration and Advanced Medical ScienceGifu University Graduate School of Medicine
Original articles

DOI: 10.1007/s00774-003-0428-4

Cite this article as:
Watanabe, N., Tezuka, Y., Matsuno, K. et al. J Bone Miner Metab (2003) 21: 344. doi:10.1007/s00774-003-0428-4

Abstract

Notch is a transmembrane protein involved in cell fate determination. In the present study, we observed temporally and spatially restricted expression of Notch1 in developing cartilage. Notch1 was localized starting from the mesenchymal condensation stage of embryonic mouse forelimbs. Interestingly, although localization could not be detected in the proliferating chondrocytes, obvious immunoreactivity indicating its expression was retained in the perichondrial region. Next, we investigated the expression of Notch1 and related molecules in a chondrogenic cell line, ATDC5 cells. Notch1, Delta-like (Dll)1, Deltex2, and Deltex3 were coexpressed after 6-day insulin treatment. Expression of Hairy and Enhancer of split homologue (HES)-1 followed thereafter. These results suggest that Notch may have a role in the early stage of chondrogenesis. To assess the effect of Notch activation, we cultured ATDC5 cells with a myeloma clone constitutively expressing Dll1, a ligand of Notch. We also used an adenovirus vector to express the constitutively active Notch1 intracellular domain (NIC). Activating either the endogenous or exogenous Notch receptor dramatically inhibited chondrogenic cell differentiation of ATDC5 cells, as assessed by Alcian blue staining of the cells and chondrocyte differentiation markers. Last, we investigated the effect of NIC on the proliferation of the ATDC5 cells. Expression of NIC by the adenovirus strongly suppressed thymidine incorporation. These results indicate that Notch is expressed in the initial stage of chondrogenic cell differentiation and has a strong inhibitory effect on both differentiation and proliferation of the cells when activated. The expression of Notch decreases as chondrogenic differentiation proceeds; however, a population of the cells with sustained expression of Notch1 become perichondrial cells. Considering that the perichondrium acts as a stem cell source of osteoblasts and chondrocytes, Notch1 may have a role in the formation of these cells by suppressing both differentiation and proliferation.

Key words

NotchchondrocyteperichondriumATDC5Deltex
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Copyright information

© Springer-Verlag Tokyo 2003