Abstract
The mitogen-activated protein (MAP) kinase family consists of extracellular signal-regulated protein kinase (ERK), p38 kinase, and c-Jun N-terminal kinase (JNK) and transduces signals from the extracellular environment to the cytoplasm and nucleus. MAP kinase signaling involves a multistep kinase cascade including MAP kinase kinase kinase (MAPKKK), MAP kinase kinase (MAPKK), and MAP kinase. The MAP kinase subtypes are constitutively expressed in articular chondrocytes and they regulate chondrocyte function, including differentiation, apoptosis, inflammatory responses, and activation of matrix metalloproteinases. Therefore, imbalance or destruction of homeostasis regulating MAP kinase activity is related to the pathogenesis of cartilage diseases such as osteoarthritis. This chapter describes methods for measuring and modulating MAP kinase subtype activity in primary cultured articular chondrocytes and cartilage explants.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Johnson, G. L. and Lapadat, R. (2002) Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298, 1911–1912.
Schaeffer, H. J. and Weber, M. J. (1999) Mitogen-activated protein kinases: specific messages from ubiquitous messengers. Mol. Cell. Biol. 19, 2435–2444.
Garrington, T. P. and Johnson, G. L. (1999) Organization and regulation of mitogen-activated protein kinase signaling pathways. Curr. Opin. Cell. Biol. 11, 211–218.
Weston, C. R. and Davis R. J. (2002) The JNK signal transduction pathway. Curr. Opin. Genet. Dev. 12, 14–21.
Kim, S.-J., Ju, J.-W., Oh, C.-D., et al.(2002) ERK-1/2 and p38 kinase oppositely regulate nitric oxide-induced apoptosis of chondrocytes in association with p53, caspase-3, and differentiation status. J. Biol. Chem. 277, 1332–1339.
Kim, S.-J., Hwang, S.-G., Shin, D.-Y., Kang, S.-S., and Chun, J.-S. (2002) p38 kinase regulates nitric oxide-induced apoptosis of articular chondrocytes by accumulating p53 via NFκ B-dependent transcription and stabilization by serine 15 phosphorylation. J. Biol. Chem. 277, 33,501–33,508.
Kim, S.-J., Kim, H.-G., Oh, C.-D., et al.(2002) p38 kinase-dependent and-independent inhibition of protein kinase C-ζ and-α regulates nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes. J. Biol. Chem. 277, 30,375–30,381.
Yoon, Y.-M., Kim, S.-J., Oh, C.-D., et al. (2002) Maintenance of differentiated phenotype of articular chondrocytes by protein kinase C and extracellular signal-regulated protein kinase. J. Biol. Chem. 277, 8412–8420.
Kim, S.-J. and Chun, J.-S. (2003) Protein kinase Cα and ζ regulate nitric oxide-induced NFκB activation that mediates cyclooxygenase-2 expression and apoptosis but not dedifferentiation in articular chondrocytes. Biochem. Biophys. Res. Commun. 303, 206–211.
Yoon, Y.-M., Kim, S.-J., Oh, C.-D., et al. (2002) Maintenance of differentiated phenotype of articular chondrocytes by protein kinase C and extracellular signal-regulated protein kinase. J. Biol. Chem. 277, 8412–8420.
Ryu, J.-H., Kim, S.-J., Kim, S.-H., et al. (2002) β-Catenin regulation of the chondrocyte phenotypes. Development 129, 5541–5550.
Kim, S.-J., Lim, D.-S., Kim, S.-H., et al. (2002) β-Catenin regulates expression of cyclooxygenase-2 in articular chondrocytes. Biochem. Biophys. Res. Commun. 296, 221–226.
Alessi, D. R., Cuenda, A., Cohen, P., Dudley, D. T., and Saltiel A. R. (1995) PD098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J. Biol. Chem. 270, 27,489–27,494.
Favata, M. F., Horiuchi, K. Y., Manos, E. J., et al. (1998) Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J. Biol. Chem. 273, 18,623–18,632.
Jiang, Y., Chen, C., Li, Z., et al. (1996) Characterization of the structure and function of a new mitogen-activated protein kinase (p38β). J. Biol. Chem. 271, 17,920–17,926.
Cuenda, A., Rouse, J., Doza, Y. N., et al. (1995) SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Lett. 364, 229–233.
Bennett, B. L., Sasaki, D. T., Murray, B. W., et al. (2001) SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc. Natl. Acad. Sci. USA 98, 13,681–13,686.
Bonny, C., Oberson, A., Negri, S., Sauser, C., and Schorderet, D. F. (2001) Cell-permeable peptide inhibitors of JNK: novel blockers of beta-cell death. Diabetes 50, 77–82.
Robbins, D. J., Zhen, E., Owaki, H., et al. (1993) Regulation and properties of extracellular signal-regulated protein kinases 1 and 2 in vitro. J. Biol. Chem. 268, 5097–5106.
Raingeaud, J., Gupta, S., Rogers, J. S., et al. (1995) Pro-inflammatory cytokines and environmental stress cause p38 mitogen-activated protein kinase activation by dual phosphorylation on tyrosine and threonine. J. Biol. Chem. 270, 7420–7426.
Raingeaud, J., Whitmarsh, A. J., Barrett, T., Derijard, B., and Davis, R. J. (1996) MKK3-and MKK6-regulated gene expression is mediated by the p38 mitogen-activated protein kinase signal transduction pathway. Mol. Cell. Biol. 16, 1247–1255.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Chun, JS. (2004). Expression, Activity, and Regulation of MAP Kinases in Cultured Chondrocytes. In: Sabatini, M., Pastoureau, P., De Ceuninck, F. (eds) Cartilage and Osteoarthritis. Methods in Molecular Medicine™, vol 100. Humana Press. https://doi.org/10.1385/1-59259-810-2:291
Download citation
DOI: https://doi.org/10.1385/1-59259-810-2:291
Publisher Name: Humana Press
Print ISBN: 978-1-58829-247-6
Online ISBN: 978-1-59259-810-6
eBook Packages: Springer Protocols