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Originally published as MBC in Press, 10.1091/mbc.E05-01-0084 on August 31, 2005

Vol. 16, Issue 11, 5316-5333, November 2005

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Microarray Analyses of Gene Expression during Chondrocyte Differentiation Identifies Novel Regulators of Hypertrophy

Claudine G. James * {dagger}, C. Thomas G. Appleton * {dagger}, Veronica Ulici * {dagger}, T. Michael Underhill * {ddagger}, and Frank Beier * {dagger} §

* CIHR Group in Skeletal Development and Remodeling, University of Western Ontario, London, Ontario N6A 5C1, Canada; {dagger} Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario N6A 5C1, Canada; {ddagger} Department of Anatomy, Cell Biology and Physiology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada; and § School of Dentistry, University of Western Ontario, London, Ontario N6A 5C1, Canada

Submitted February 1, 2005; Revised June 24, 2005; Accepted August 24, 2005
Monitoring Editor: Marianne Bronner-Fraser

Ordered chondrocyte differentiation and maturation is required for normal skeletal development, but the intracellular pathways regulating this process remain largely unclear. We used Affymetrix microarrays to examine temporal gene expression patterns during chondrogenic differentiation in a mouse micromass culture system. Robust normalization of the data identified 3300 differentially expressed probe sets, which corresponds to 1772, 481, and 249 probe sets exhibiting minimum 2-, 5-, and 10-fold changes over the time period, respectively. GeneOntology annotations for molecular function show changes in the expression of molecules involved in transcriptional regulation and signal transduction among others. The expression of identified markers was confirmed by RT-PCR, and cluster analysis revealed groups of coexpressed transcripts. One gene that was up-regulated at later stages of chondrocyte differentiation was Rgs2. Overexpression of Rgs2 in the chondrogenic cell line ATDC5 resulted in accelerated hypertrophic differentiation, thus providing functional validation of microarray data. Collectively, these analyses provide novel information on the temporal expression of molecules regulating endochondral bone development.

This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E05-01-0084) on August 31, 2005.

Address correspondence to: Frank Beier (fbeier{at}uwo.ca).




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