Avian neural crest cell attachment to laminin: involvement of divalent cation dependent and independent integrins.
The mechanisms of neural crest cell interaction with laminin were explored using a quantitative cell attachment assay. With increasing substratum concentrations, an increasing percentage of neural crest cells adhere to laminin. Cell adhesion at all substratum concentrations was inhibited by the CSAT antibody, which recognizes the chick beta 1 subunit of integrin, suggesting that beta 1-integrins mediate neural crest cell interactions with laminin. The HNK-1 antibody, which recognizes a carbohydrate epitope, inhibited neural crest cell attachment to laminin at low coating concentrations (greater than 1 microgram ml-1; Low-LM), but not at high coating concentration of laminin (10 micrograms ml-1; High-LM). Attachment to Low-LM occurred in the absence of divalent cations, whereas attachment to High-LM required greater than 0.1 mM Ca2+ or Mn2+. Neural crest cell adherence to the E8 fragment of laminin, derived from its long arm, was similar to that on intact laminin at high and low coating concentrations, suggesting that this fragment contains the neural crest cell binding site(s). The HNK-1 antibody recognizes a protein of 165,000 Mr which is also found in immunoprecipitates using antibodies against the beta 1 subunit of integrin and is likely to be an integrin alpha subunit or an integrin-associated protein. Our results suggest that the HNK-1 epitope on neural crest cells is present on or associated with a novel or differentially glycosylated form of beta 1-integrin, which recognizes laminin in the apparent absence of divalent cations. We conclude that neural crest cells have at least two functionally independent means of attachment to laminin which are revealed at different substratum concentrations and/or conformations of laminin.
Lallier
T
T
Developmental Biology Center, University of California, Irvine 92717.
Bronner-Fraser
M
M
D000818:Animals; D000943:Antigens, Differentiation / Q000276:immunology; D018959:CD57 Antigens; D002413:Cations, Divalent; D002448:Cell Adhesion / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003370:Coturnix; D005109:Extracellular Matrix / Q000502:physiology; D005353:Fibronectins / Q000502:physiology; D016023:Integrins / Q000276:immunology / Q000502:physiology*; D007797:Laminin / Q000502:physiology*; D009432:Neural Crest / Q000166:cytology / Q000502:physiology*; D011233:Precipitin Tests
D000943:Antigens, Differentiation; D018959:CD57 Antigens; D002413:Cations, Divalent; D005353:Fibronectins; D016023:Integrins; D007797:Laminin