Neurite extension by neuroblastoma cells on substratum-bound fibronectin's cell-binding fragment but not on the heparan sulfate-binding protein, platelet factor-4.
Human and rat neuroblastoma cells extend neurites over plasma fibronectin (pFN)-coated substrata. For resolution of which fibronectin binding activities (the cell-binding domain (CBD), the heparan sulfate-binding domains, or a combination of the two) are responsible for neurite outgrowth, CBD was prepared free of heparan sulfate-binding activity as described by Pierschbacher et al. (Cell 26 (1981) 259-267). Neuroblastoma cells attached and extended neurites as stably and as effectively on CBD-coated substrata as on intact pFN, while cytoplasmic spreading was more extensive on pFN-coated substrata. The structures of growth cones on CBD or pFN were virtually identical. On substrata coated with the model heparan sulfate-binding protein, platelet factor 4 (PF4), cells attached and spread somewhat but never extended neurites. When cells were challenged with substrata coated with various ratios of CBD and PF4, PF4 was found to be an effective inhibitor of CBD-mediated neurite extension. Similarly, cells grown on substrata coated at different locations with CBD or PF4 in order to evaluate topographical dependence of growth cone formation extended neurites only onto the CBD-coated region or along the interface between these two proteins, but never onto the PF4 side of cells that bridged the interface. These studies indicate that (a) the CBD activity of pFN, and not its heparan sulfate-binding activity, is the critical determinant in neurite extension of these neural tumor cells from the central nervous system; (b) under some circumstances, heparan sulfate-binding activity can be antagonistic to neurite extension; (c) the chemical nature of the substratum controls the direction of neurite extension; (d) these neuroblastoma cells respond to these binding proteins very differently than fibroblasts or neurons from the peripheral nervous system.
Tobey
S L
SL
McClelland
K J
KJ
Culp
L A
LA
D000818:Animals; D002448:Cell Adhesion; D002918:Chymotrypsin / Q000378:metabolism; D004533:Egtazic Acid; D005353:Fibronectins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D009447:Neuroblastoma / Q000648:ultrastructure*; D009474:Neurons / Q000166:cytology*; D010978:Platelet Factor 4 / Q000378:metabolism*; D051381:Rats
D002918:Chymotrypsin; D004533:Egtazic Acid; D005353:Fibronectins; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D010978:Platelet Factor 4