Reversible suppression by ionophore A23187 of retinoic acid-induced cartilage resorption in cultured fetal rat bones

  • Andreas Kistler
Article

Summary

In fetal rat bones in culture the divalent cation ionophore A23187 inhibited in a dose-dependent manner both the release of proteoglycans and the subsequent degradation of cartilage induced by retinoic acid, indicating that calcium was involved in its action. A23187 had to be present continuously to manifest its inhibitory effect; retrieval of the ionophore abolished the suppression, demonstrating that the effect was reversible and not due to toxicity. A23187 at 1.0 μM, which completely blocked the retinoic acid-induced cartilage resorption, markedly suppressed3H-leucine,3H-mannose and3H-thymidine incorporation in control and retinoic acid-treated cultures. Reduced3H-thymidine incorporation did not appear to be responsible for the inhibition by A23187 of retinoic acid-induced cartilage resorption because inhibitors of DNA synthesis did not affect the retinoic acid response. In the presence of retinoic acid the ionophore at 0.3 μM had no effect on the incorporation of3H-leucine and3H-mannose, but suppressed the retinoic acid-induced proteoglycan release. This suggests that reduced protein and glycoprotein synthesis were not the main causes for the inhibitory effect of A23187. In conclusion, retinoic acid-induced cartilage degradation required calcium at some crucial points.

Key words

Retinoic acid Cartilage resorption Bone culture Ionophore A23187 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Berridge MJ (1975) The interaction of cyclic nucleotides and calcium in the control of cellular activity. In: Greengard P, Robison A (eds) Advances in cyclic nucleotide research, vol 6. Raven Press, New York, pp 1–98Google Scholar
  2. Bollag W (1979) Retinoids and cancer. Cancer Chemother Pharmacol 3:207–215PubMedGoogle Scholar
  3. Breitman TR, Selonick ST, Collins SJ (1980) Induction of differentiation of the human promyelocytic leukemia cell line (HL-60) by retinoic acid. Proc Natl Acad Sci USA 77:2936–2940PubMedGoogle Scholar
  4. Cheung WY (1980) Calmodulin plays a pivotal role in cellular regulation. Science 207:19–27PubMedGoogle Scholar
  5. Chytil F, Ong DE (1976) Mediation of retinoic acid-induced growth and anti-tumour activity. Nature 260:49–51PubMedGoogle Scholar
  6. De Luca LM, Yuspa SH (1974) Altered glycoprotein synthesis in mouse epidermal cells treated with retinyl acetate in vitro. Exp Cell Res 86:106–110PubMedGoogle Scholar
  7. Dhouailly D, Hardy MH, Sengel P (1980) Formation of feathers on chick foot scales: a stage-dependent morphogenetic response to retinoic acid. J Embryol Exp Morphol 58:63–78PubMedGoogle Scholar
  8. Dietrich JW, Paddock DN (1979) In vitro effects of ionophore A23187 on skeletal collagen and noncollagen protein synthesis. Endocrinology 104:493–499PubMedGoogle Scholar
  9. Dziak R, Stern PH (1975) Calcium transport in isolated bone cells III. Effects of parathyroid hormone and cyclic 3′,5′-AMP. Endocrinology 97:1281–1287PubMedGoogle Scholar
  10. Dziak R, Stern PH (1976) Responses of fetal rat bone cells and bone organ cultures to the ionophore, A23187. Calcif Tiss Int 22:137–147Google Scholar
  11. Fell HB, Mellanby E (1952) The effect of hypervitaminosis A on embryonic limb-bones cultivated in vitro. J Physiol 116:320–349PubMedGoogle Scholar
  12. Gallandre F, Kistler A, Galli B (1980) Inhibition and reversion of chondrogenesis by retinoic acid in rat limb bud cell cultures. Wilhelm Roux's Arch 189:25–33Google Scholar
  13. Gillies RJ (1982) Calcium, calmodulin and cAMP and the control of cellular proliferation. Trends Biochem Sci 7:233–235Google Scholar
  14. Hardy MH (1968) Glandular metaplasia of hair follicles and other responses to vitamin A excess in cultures of rodent skin. J Embryol Exp Morphol 19:157–180PubMedGoogle Scholar
  15. Heisler S (1976) Calcium and cyclic nucleotide involvement in exocrine pancreatic enzyme secretion studied with the ionophore A23187. Life Sci 19:233–242PubMedGoogle Scholar
  16. Jetten AM, Jetten MER (1979) Possible role of retinoic acid binding protein in retinoid stimulation of embryonal carcinoma cell differentiation. Nature 278:180–182PubMedGoogle Scholar
  17. Kistler A (1978) Inhibition of vitamin A action in rat bone cultures by inhibitors of RNA and protein synthesis. Experientia 34:1159–1161PubMedGoogle Scholar
  18. Kistler A (1982) Retinoic acid-induced cartilage resorption. Induction of specific changes in protein synthesis and inhibition by tunicamycin. Differentiation 21:168–174PubMedGoogle Scholar
  19. Kistler A, Galli B (1979) Retinoic acid induced proteoglycan release and cartilage resorption in rat bone cultures are age dependent and inhibited by EDTA. Wilhelm Roux's Arch 187:59–71Google Scholar
  20. Kistler A, Hartmann HR (1980) Requirement of RNA and protein synthesis and inhibition by ethylenediaminetetraacetic acid of retinoic acid induced proteoglycan release in a transplantable rat chondrosarcoma. J Natl Cancer Inst 64:625–630Google Scholar
  21. Lewis CA, Pratt RM, Pennypacker JP, Hassell JR (1978) Inhibition of limb chondrogenesis in vitro by vitamin A: alterations in cell surface characteristics. Dev Biol 64:31–47PubMedGoogle Scholar
  22. Lorenzo JA, Raisz LG (1981) Divalent cation ionophores stimulate resorption and inhibit DNA synthesis in cultured fetal rat bone. Science 212:1157–1159PubMedGoogle Scholar
  23. Lotan R (1980) Effects of vitamin A and its analogs (retinoids) on normal and neoplastic cells. Biochim Biophys Acta 605:33–91PubMedGoogle Scholar
  24. Lowry OH, Rosebrough NS, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  25. Moore T (1967) Effects of vitamin A deficiency in animals. Pharmacology and toxicology of vitamin A. In: Sebrell WH, Harris RS (eds) The vitamins, vol I. Academic Press, New York, pp 245–266Google Scholar
  26. Sidell N (1982) Retinoic acid-induced growth inhibition and morphologic differentiation of human neuroblastoma cells in vitro. J Natl Cancer Inst 68:581–593Google Scholar
  27. Stern PH (1977) Ionophores: chemistry, physiology and potential applications to bone biology. Clin Orthop 122:273–298PubMedGoogle Scholar
  28. Stern PH, Orr MF, Brull E (1982) Ionophore A23187 promotes osteoclast formation in bone organ culture. Calcif Tissue Int 34:31–36PubMedGoogle Scholar
  29. Strickland S, Mahdavi V (1978) The induction of differentiation in teratocarcinoma stem cells by retinoic acid. Cell 15:393–404CrossRefPubMedGoogle Scholar
  30. Whiteman P (1973) The quantitative measurement of alcian blue-glycosaminoglycan complexes. Biochem J 131:343–350PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Andreas Kistler
    • 1
  1. 1.Biological Pharmaceutical Research DepartmentF. Hoffmann-La Roche & Co. Ltd.BasleSwitzerland

Personalised recommendations