The mechanism of precartilage mesenchymal condensation: a major role for interaction of the cell surface with the amino-terminal heparin-binding domain of fibronectin.
Using low magnification Hoffman Modulation Contrast microscopy to rapidly identify precartilage mesenchymal condensations in chick limb bud cultures, we have determined the effect on condensation number of treatments disruptive of the interaction of cell surface components with endogenously produced fibronectin. A monoclonal antibody directed against the amino-terminal heparin-binding domain of fibronectin reduced the number of condensations by more than 50%, as did the oligopeptide gly-arg-gly, which is a repeated motif in that fibronectin domain. In contrast, monoclonal antibodies directed against the collagen- and integrin-binding domains of fibronectin, or oligopeptides containing the fibronectin integrin-recognition sequence arg-gly-asp-ser, had no significant effect on condensation number. Addition of Flavobacterium heparinase to cultures also reduced condensation number by more than 50%. Alcian blue staining of sulfated proteoglycan was greatly reduced in differentiated cultures that had been exposed to treatments that reduced condensation number. Taken together with the accompanying study, which directly demonstrates an adhesive interaction between the amino-terminal domain of extracellular fibronectin and heparin-like molecules on the surfaces of latex bead probes, the data presented here strongly indicate a major role for the corresponding cell-matrix interaction in mediating precartilage condensation in limb mesenchyme.
Frenz
D A
DA
Department of Cell Biology and Anatomy, New York Medical College, Valhalla 10595.
Jaikaria
N S
NS
Newman
S A
SA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002356:Cartilage / Q000196:embryology*; D002448:Cell Adhesion; D002462:Cell Membrane / Q000502:physiology*; D002478:Cells, Cultured; D002642:Chick Embryo; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D019763:Heparin Lyase; D008648:Mesoderm / Q000502:physiology*; D008969:Molecular Sequence Data; D009024:Morphogenesis; D009842:Oligopeptides / Q000494:pharmacology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism*; D014921:Wings, Animal / Q000196:embryology
D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans; D005353:Fibronectins; D009842:Oligopeptides; D011509:Proteoglycans; D006493:Heparin; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase