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Double-stranded RNA-dependent protein kinase regulates insulin-stimulated chondrogenesis in mouse clonal chondrogenic cells, ATDC-5

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Abstract

Double-stranded RNA-dependent protein kinase (PKR) is an interferon-induced protein that has been identified and characterized as a translational inhibitor in an interferon-regulated antiviral pathway. PKR is also reported to play important roles in the regulation of cell growth and differentiation. We have previously demonstrated that PKR inactivation suppresses osteoblast calcification and osteoclast formation. However, reports concerning the roles of PKR in chondrogenesis are limited. In this study, we have demonstrated that PKR is required for the in vitro differentiation of the mouse clonal chondrogenic cell line ATDC-5. ATDC-5 cells treated with insulin differentiated into chondrocytes and produced an alcian-blue-positive cartilage matrix. The protein expression of signal transducers and activators of transcription (STAT) peaked at day 7 of differentiation, whereas the expression of SRY-box-containing gene 9 (Sox-9), which is a transcription factor for chondrocyte differentiation, increased gradually. When the cells were treated with a PKR inhibitor (2-aminopurine), the cartilage matrix formation decreased. The protein expression of STAT1 continued to increase up to day 21, whereas the expression of Sox-9 was low and did not increase. We also demonstrated that PKR was localized to a marginal region of the mandibular condyle cartilage in mouse embryos. Our findings suggest that PKR has important functions in the differentiation of chondrocytes through the modulation of STAT1 and Sox-9 expression.

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Abbreviations

2-AP:

2-Aminopurine

STAT1:

Signal transducers and activators of transcription 1

OSX:

Osterix

Sox-9:

SRY-box-containing gene 9

ITS:

Insulin, transferrin and selenium

PKR:

Double-stranded RNA-dependent protein kinase

eIF-2α:

α-Subunit of eukaryotic initiation factor 2

TNF-α:

Tumor necrosis factor-α

IL-1:

Interleukin-1

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Acknowledgments

We thank Ms. Toyono Nobukuni for her skillful technical assistance.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Anatomy, School of Medicine, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi, Kitakyushu, Fukuoka, 807-8555, Japan

    Hiroyuki Morimoto, Ryoko Baba & Yoshiaki Doi

  2. Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima, Graduate School, 3-18-15, Kuramoto, Tokushima, 770-8504, Japan

    Tatsuji Haneji

Authors
  1. Hiroyuki Morimoto
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  2. Ryoko Baba
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  3. Tatsuji Haneji
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  4. Yoshiaki Doi
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Corresponding author

Correspondence to Hiroyuki Morimoto.

Additional information

This work is dedicated to the late Dr. Yoshiaki Doi.

This work was supported in part by a Grant-in-Aid for Scientific Research (KAKENHI) from the Ministry of Education Culture, Sports Science and Technology of Japan (grants 22592055 [to H.M.], 22700770 [to R.B.], 20515986 [to T.H.] and 23590250 [to Y.D.]).

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Morimoto, H., Baba, R., Haneji, T. et al. Double-stranded RNA-dependent protein kinase regulates insulin-stimulated chondrogenesis in mouse clonal chondrogenic cells, ATDC-5. Cell Tissue Res 351, 41–47 (2013). https://doi.org/10.1007/s00441-012-1521-6

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  • DOI: https://doi.org/10.1007/s00441-012-1521-6

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