Skip to main content
Log in

Apoptosis staining in cultured pseudoachondroplasia chondrocytes

  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Pseudoachondroplasia (PSACH) is a skeletal dysplasia caused by a mutation in cartilage oligomeric matrix protein (COMP), a glycoprotein of normal cartilage matrix. PSACH chondrocytes have a distinctive phenotype with enlarged rER cisternae containing COMP, aggrecan, type IX collagen, and chaperone proteins. Ultrastructural studies suggested that this accumulation compromises cell function, hastening cell death, and consequently reducing the number of cells in the growth plate contributing to linear bone growth. Using the alginate bead system, we cultured control and PSACH chondrocytes for twenty weeks and one year to determine the effect of the mutation on size and number of cartilage nodules; and the presence of apoptotic cell death (TUNEL assay). At 20 weeks, beads containing PSACH or control chondrocytes did not differ in size and number of cartilage nodules or number of TUNEL-positive cells. After one year, nodule number, size and percent cartilage per bead were significantly less in PSACH nodules, and the number of cells staining positive for apoptosis was significantly greater than in controls (71.8% vs. 44.6%). The increase in apoptosis in PSACH nodules correlates with a decrease in growth of cartilage, supporting our hypothesis that death of damaged cells contributes to the growth plate defects in PSACH.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Hecht JT, Nelson LD, Crowder E, et al. Mutations in exon 17B of cartilage oligomeric matrix protein (COMP) cause pseudoachondroplasia. Nature Genet 1995; 10: 325–329.

    Google Scholar 

  2. Briggs MD, Mortier GR, Cole WG, et al. Diverse mutations in the gene for cartilage oligomeric matrix protein in Pseudoachondroplasia—Multiple Epiphyseal Dysplasia spectrum. Am J Human Genet 1995; 62: 311–319.

    Google Scholar 

  3. Cooper RR, Poinsetti IV, Maynard JA. A rough surfaced endoplasmic reticulum storage disorder. J Bone Joint Surg 1973; 55A: 475–484.

    Google Scholar 

  4. Stanescu V, Maroteaux P, Stanescu R. The biochemical defect of pseudoachondroplasia. Eur J Pediatr 1982; 138: 221–225.

    Google Scholar 

  5. Hecht JT, Montufar-Solis D, Decker G, et al. Retention of cartilage oligomeric matrix protein (COMP) and cell death in redifferentiated chondrocytes. Matrix Biol 1998; 17: 625–633.

    Google Scholar 

  6. Hecht JT, Hayes E, Snuggs M, et al. Calreticulin, PDI, Grp94 and BiP chaperone proteins are associated with retained COMP in psuedoachondroplasia chondrocytes. Matrix Biol 2001; 20: 251–262.

    Google Scholar 

  7. Ashkenazi A, Dixit VM. Apoptosis control by death and decoy receptors. Current Opinion Cell Biol 1999; 11: 255–260.

    Google Scholar 

  8. Horton JR, Feng WE, L., Adams C. Chondrocyte apoptosis in development, aging and disease. Matrix Biol 1998; 17: 107–115.

    Google Scholar 

  9. Gibson G. Active role of chondrocyte apoptosis in endochondral ossification. Microsc Res Tech 1998; 43: 191–204.

    Google Scholar 

  10. Bossy-Wetzel E, Green DR. Apoptosis: Checkpoint at the mitochondrial frontier. Mutat Res 1999; 434: 243–251.

    Google Scholar 

  11. Wang J, Lenardo MJ. Roles of caspases in apoptosis, development and cytokine maturation revealed by homozygous gene deficiencies. J Cell Sci 2000; 113: 753–757.

    Google Scholar 

  12. Nagata S. Apoptotic DNA fragmentation. Exp Cell Res 2000; 256: 12–18.

    Google Scholar 

  13. Duriez PJ, Shah GM. Cleavage of poly(ADP-ribose) polymerase: A sensitive parameter to study cell death. Biochem Cell Biol 1997; 75: 337–349.

    Google Scholar 

  14. Kothakota S, Azuma T, Reinhard C, et al. Caspase-3-generated fragment of gelsolin: Effector of morphological change in apoptosis. Science 1997; 278: 294–298.

    Google Scholar 

  15. Koopman G, Reutelingsperger CPM, Kuijten GAM, et al. Annexin V for flow cytometric detection of phosphatidylserine expression in B cells undergoing apoptosis. Blood 1994; 84: 1415–1420.

    Google Scholar 

  16. Wang Y, Toury R, Hauchecorne M, Blamain N. Expression of Bcl-2 protein in the epiphyseal plate cartilage and trabecular bone of growing rats. Histochem Cell Biol 1997; 108: 45–55.

    Google Scholar 

  17. Amling M, Neff L, Tanaka S, et al. Bcl-2 lies downstream of parathyroid hormone-related peptide in a signaling pathway that regulates chondrocyte maturation during skeletal development. J Cell Biol 1997; 136: 205–213.

    Google Scholar 

  18. Hauselmann HJ, Aydelotte MB, Kuettner KE, et al. Synthesis and turnover of proteoglycans by human and bovine articular chondrocytes cultured in alginate beads. Matrix 1992; 12: 116–129.

    Google Scholar 

  19. Holtzer H, Abbott J, Lah J, Holtzer S. The loss of phenotypic traits by dedifferentiated cells in vitro. I. Dedifferentiation of cartilage cells. Proc Natl Acad Sci USA 1960; 49: 643–647.

    Google Scholar 

  20. Erenpreisa J, Roach HI. Aberrant death in dark chondrocytes of the avian growth plate. Cell Death Differentiation 1998; 5: 60–66.

    Google Scholar 

  21. Bronckers AL, Goei W, Luo G, et al. DNA fragmentation during bone formation in neonatal rodents assessed by transferasemediated end labeling. J Bone Min Res 1996; 11: 1281–1291.

    Google Scholar 

  22. Adams CS, Mansfield KD, Rajpurohit R, et al. Components of the cartilage extracellular matrix regulate chondrocyte apoptosis. In Shapiro IM, Boyan B, Anderson HC, eds. The Growth Plate. Amsterdam: IOS Press 2002: 63–72.

    Google Scholar 

  23. Gibson G, Wang X, Yang M. The release and activation of TGFB associated with chondrocyte hypertrophy and apoptosis. In Shapiro IM, Boyan B, Anderson HC, eds. The Growth Plate. Amsterdam: IOS Press 2002: 77–92.

    Google Scholar 

  24. Roach HI. Cell death and transdifferentiation in the growth plate. In Shapiro IM, Boyan B, Anderson HC, eds. The Growth Plate. Amsterdam: IOS Press 2002: 93–104.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Duke, J., Montufar-Solis, D., Underwood, S. et al. Apoptosis staining in cultured pseudoachondroplasia chondrocytes. Apoptosis 8, 191–197 (2003). https://doi.org/10.1023/A:1022926811397

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022926811397

Navigation