Specific effects of retinoic acid on the skeletal morphogenesis of the 11‐day mouse embryo forelimb bud in vitro

Authors:
  • LMLucette Meunier
    Lucette Meunier
  • BLBernard Lassalle
    Bernard Lassalle
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Abstract
Summary— Using our improved method for culturing 11‐day mouse forelimb buds in vitro, we have investigated the effects of a local application of all‐trans‐retinoic acid (RA) on growth, cartilaginous differentiation and skeletal patterning in the mammalian limb bud. Carrier implants of catgut impregnated with DMSO or various doses of RA in DMSO were inserted at the apex of the buds in the proximo‐distal axis just beneath the apical ectodermal ridge. After 6 days of culture, cartilaginous skeletons were stained and explants were processed for morphological analysis and quantitative study using computerized optical image analysis. Buds treated with low doses of RA exhibited stimulated growth and chondrogenesis. Moreover, hypertrophied and fused metacarpals were seen within explants treated with the lowest dose. High doses strongly inhibited growth and skeletal morphogenesis. An intermediate dose sustained cartilaginous differentiation at the same level as low doses, but concomitantly disturbed the skeletal pattern. These results are discussed considering reported RA effects on other experimental systems including avian limb bud as an in vivo model or cell cultures as an in vitro simplified model.

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