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Inhibition and reversion of chondrogenesis by retinoic acid in rat limb bud cell cultures

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Summary

Mesenchyme cells derived from embryonic rat limb buds cultured at high density differentiated into chondrocytes. The degree of chondrogenesis was assessed by alcian blue staining, a stain specific for cartilage matrix. The addition of retinoic acid on day 1 of culture inhibited chondrogenesis in a dose-dependent fashion. When retinoic acid was added to the cultures on day 5, the cartilage nodules, consisting of newly differentiated cartilage cells, disappeared during the following 6 days. Coinciding with this process the histochemically demonstrable alkaline phosphatase activity, localized in the internodular areas, also disappeared. This indicated that retinoic acid not only inhibited chondrogenesis but also induced resorption of cartilage cells and that at least two cell types were affected, the cartilage cells and the cells bearing alkaline phosphatase.

Actinomycin D and cycloheximide, inhibitors of RNA and protein synthesis, suppressed the retinoic acid effect in day 5 limb bud cell cultures. This result indicated that the effect of retinoic acid required RNA and protein synthesis and is compatible with the view that vitamin A may act in a hormone-like way.

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References

  • Blalock JE, Gifford GE (1977) Retinoic acid (vitamin A acid) induced transcriptional control of interferon production. Proc Natl Acad Sci USA 74:5382–5386

    Google Scholar 

  • Burton K (1956) A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 62:315–323

    Google Scholar 

  • Cabrini RL (1961) Histochemistry of ossification. Int Rev Cytol 11:283–306

    Google Scholar 

  • De Luca LM, Yuspa SH (1974) Altered glycoprotein synthesis in mouse epidermal cells treated with retinyl acetate in vitro. Exp Cell Res 86:106–110

    Google Scholar 

  • Dhouailly D, Hardy MH (1978) Retinoic acid causes development of feathers in scale-forming integument of chick embryo. Wilhelm Roux's Archives 185:195–200

    Google Scholar 

  • Dion LD, Blalock JE, Gifford GE (1977) Vitamin A-induced density-dependent inhibition of L-cell proliferation. J Nat Cancer Inst 58:795–801

    Google Scholar 

  • Dion LD, Blalock JE, Gifford GE (1978) Retinoic acid and the restoration of anchorage dependent growth to transformed mamalian cells. Exp Cell Res 117:15–22

    Google Scholar 

  • Fell HB (1957) The effect of excess vitamin A on cultures of embryonic chick skin explanted at different stages of differentiation. Proc R Soc London Ser B: 146:242–256

    Google Scholar 

  • Fell HB, Mellanby E (1952) The effect of hypervitaminosis A on embryonic limb-bones cultivated in vitro. J Physiol 116:320–349

    Google Scholar 

  • Hardy MH (1968) Glandular metaplasia of hair follicles and other responses to vitamin A excess in cultures of rodent skin. J Embryol Exp Morphol 19:157–180

    Google Scholar 

  • Hassell JR, Pennypacker JP, Lewis CA (1978) Chondrogenesis and cell proliferation in limb bud cell cultures treated with cytosine arabinoside and vitamin A. Exp Cell Res 112:409–417

    Google Scholar 

  • Jetten AM, Jetten MER (1979) Possible role of retinoic acid binding protein in retinoid stimulation of embryonal carcinoma cell differentiation. Nature 278:180–182

    Google Scholar 

  • Kistler A (1978) Inhibition of vitamin A action in rat bone cultures by inhibitors of RNA and protein synthesis. Experientia 34:1159–1161

    Google Scholar 

  • 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 Archives 187:59–71

    Google Scholar 

  • Kistler A, Hartmann HR (1980) Requirement of RNA and protein synthesis and inhibition by EDTA of retinoic acid-induced proteoglycan release in a transplantable rat chondrosarcoma. J Nat Cancer Inst 64:625–630

    Google Scholar 

  • Kochhar DM (1973) Limb development in mouse embryos. I. Analysis of teratogenic effects of retinoic acid. Teratology 7:289–298

    Google Scholar 

  • Lasnitzki I (1955) The effect of excess vitamin A on mitosis in chick heart fibroblasts in vitro. Exp Cell Res 8:121–125

    Google Scholar 

  • 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 64:31–47

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Merriman RL, Bertram JS (1979) Reversible inhibition by retinoids of 3-methylcholanthrene-induced neoplastic transformation in C3H/10T1/2 clone cells. Cancer Res 39:1661–1666

    Google Scholar 

  • Ong DE, Chytil F (1975) Retinoic acid-binding protein in rat tissue. Partial purification and comparison to rat tissue retinol-binding protein. J Biol Chem 250:6113–6117

    Google Scholar 

  • Pearse AGE (1968) Alkaline phosphatases. In: Histochemistry, 3rd edn. Theoretical and applied, vol 1. Churchill Livingstone, Edinburgh London New York, pp 495–546

    Google Scholar 

  • Pennypacker JP, Lewis CA, Hassell JR (1978) Altered proteoglycan metabolism in mouse limb mesenchyme cell cultures treated with vitamin A. Arch Biochem Biophys 186:351–358

    Google Scholar 

  • Sani BP, Hill DL (1974) Retinoic acid: A binding protein in chick embryo metatarsal skin. Biophys Res Commun 61:1276–1282

    Google Scholar 

  • Shapiro SS, Poon JP (1976) Effect of retinoic acid on chondrocyte glycosaminoglycan biosynthesis. Arch Biochem Biophys 174:74–81

    Google Scholar 

  • Solursh M, Meier S (1973) The selective inhibition of mucopolysaccharide synthesis by vitamin A treatment of cultured chick embryo chondrocytes. Calcif Tissue Res 13:131–142

    Google Scholar 

  • Strickland S, Mahdavi V (1978) The induction of differentiation in teratocarcinoma stem cells by retinoic acid. Cell 15:393–404

    Google Scholar 

  • Vasan NS, Lash JW (1975) Chondrocyte metabolism as affected by vitamin A. Calcif Tissue Res 19:99–107

    Google Scholar 

  • Yuspa SH, Harris CC (1974) Altered differentiation of mouse epidermal cells treated with retinyl acetate in vitro. Exp Cell Res 86:95–105

    Google Scholar 

  • Zimmermann B (1978) The development of alkaline phosphatase activity in limb buds of mouse embryos in vitro and its relation to chondrogenesis. Anat Embryol Z Anat Entwicklungsgesch 153:95–104

    Google Scholar 

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Gallandre, F., Kistler, A. & Galli, B. Inhibition and reversion of chondrogenesis by retinoic acid in rat limb bud cell cultures. Wilhelm Roux' Archiv 189, 25–33 (1980). https://doi.org/10.1007/BF00848564

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  • DOI: https://doi.org/10.1007/BF00848564

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