Differential outgrowth of retinal neurites on purified extracellular matrix molecules.
Organotypic cultures of the embryonic retina were used to study the influence of extracellular matrix molecules on neurite elongation during development of the central nervous system. Microexplants from the chick retina (embryonic day 6) were grown in medium containing appropriate trophic support on purified matrix molecules adsorbed to plastic at various concentrations. The maximum neurite length obtained on each type of substratum was measured on day 4 of culture. No fiber outgrowth occurred on substrata of vitronectin or a hyaluronate-binding chondroitin sulfate proteoglycan. In contrast, neurite elongation was strongly promoted on laminin in a dose-dependent manner. Fibronectin elicited a neurite outgrowth corresponding to about one-third the length of the outgrowth on laminin. A 31,000-dalton fibronectin fragment representing the heparin-binding domain elicited neurite elongation comparable to that promoted by the intact fibronectin molecule. Other isolated domains of fibronectin, including the 105,000-dalton "cell-binding" domain, did not allow neurite outgrowth. Furthermore, preincubation of fibronectin substratum with antibodies to the heparin-binding fibronectin fragment entirely prevented outgrowth. Fiber outgrowth was also evoked on substrata of platelet factor 4, a protein binding heparan sulfate. Adding increasing concentrations of heparin progressively inhibited the neurite extension on laminin, whereas similar addition of soluble chondroitin sulfate proteoglycan had no effect. The results indicate that growing retinal neurites show strong preference for laminin versus fibronectin. Moreover, the outgrowth-promoting activity of both cell adhesion proteins seems to be localized to their heparin-binding regions. It is suggested that during development of the visual system, elongating retinal neurites can actively discriminate between different extracellular molecules by a mechanism that may involve participation of cell surface heparan sulfate proteoglycans.
Carri
N G
NG
Department of Zoology, Uppsala University, Sweden.
Perris
R
R
Johansson
S
S
Ebendal
T
T
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D046508:Culture Techniques / Q000379:methods; D003712:Dendrites / Q000187:drug effects / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000494:pharmacology*; D006023:Glycoproteins / Q000494:pharmacology; D006025:Glycosaminoglycans / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D007797:Laminin / Q000494:pharmacology*; D010446:Peptide Fragments / Q000494:pharmacology; D012160:Retina / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D013997:Time Factors; D019096:Vitronectin
D005353:Fibronectins; D006023:Glycoproteins; D006025:Glycosaminoglycans; D007797:Laminin; D010446:Peptide Fragments; D019096:Vitronectin; D006493:Heparin