Heparan sulfate in the inner limiting membrane of embryonic chicken retina binds basic fibroblast growth factor to promote axonal outgrowth.
During neural development retinal ganglion cell axons migrate over the retinal basal lamina (inner limiting membrane, ILM) in directed growth toward the optic nerve. We found that both growth rate and distribution density of the ganglion cell axons on isolated cell-free ILM was greatly inhibited by pretreatment with heparitinase but not with chondroitinase ABC. The persistence of radioactively labeled proteoglycans added to the culture medium eliminated residual heparitinase as an explanation for the inhibition. A cell binding assay showed that heparitinase acted on the ILM to influence axonal behavior without apparent inhibition of cell adhesion. These results indicated that the neurite outgrowth promoting activity of the ILM depended on the heparan sulfate (HS) side chains of its proteoglycans. Basic fibroblast growth factor (bFGF) stimulated additional neuronal sprouting and neurite elongation on the ILM. This neurotropic activity of bFGF was inhibited by heparitinase pretreatment of the ILM, suggesting that bFGF bound to HS on the ILM. The activity of bFGF was enhanced by exogenous heparin added to the culture medium; although heparin alone failed to stimulate either neurite extension or neuronal cell sprouting. These results demonstrate that HS in the ILM possesses neurotropic activity for axons of the ganglion cells by binding bFGF for presentation to cell-surface receptors and may, therefore, play a significant role in stimulating axonal outgrowth during development.
Chai
L
L
Department of Zoology, Oregon State University, Corvallis 97331, USA.
Morris
J E
JE
D000818:Animals; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002642:Chick Embryo; D019765:Chondroitin ABC Lyase / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism* / Q000494:pharmacology; D006031:Glycosylation; D006493:Heparin / Q000494:pharmacology; D006497:Heparan Sulfate / Q000378:metabolism*; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism*; D012160:Retina / Q000166:cytology / Q000196:embryology*; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000648:ultrastructure*; D015398:Signal Transduction
D019765:Chondroitin ABC Lyase; D016222:Fibroblast Growth Factor 2; D006493:Heparin; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D017468:Receptors, Fibroblast Growth Factor; C021349:heparitinsulfate lyase