Functional interactions of neuronal heparan sulphate proteoglycans with laminin

Quantitative biosynthetic studies using cellular extracts and neuron conditioned medium demonstrated that heparan sulphate proteoglycans (HSPGs) comprised 20-25% of the sulphated proteoglycans produced by neurons while the remainder consisted of chondroitin sulphate proteoglycans (CSPGs). When chromatographic fractions containing guanidine extracted and partially purified proteoglycans from culture medium conditioned by neurons (NCM) were used to pretreat a laminin substrate, neurite formation by sensory neurons was enhanced. Enhanced neurite promoting activity was not apparent if, during the pretreatment of the laminin substrate with NCM, heparan sulphate glycosaminoglycans (HS) were present. To determine the molecular basis of cell surface HSPG interactions with immobilized laminin, adhesion and neurite growth by dissociated sensory neurons were quantified at 4 h in vitro--a time at which there was no apparent contribution of released proteoglycans to neurite growth. Whereas adhesion was not influenced, neurite growth was partially inhibited in a dose-dependent manner if the sensory neurons were coincubated with HS, and if the cells were pretreated, prior to seeding, with heparitinase. The inhibitory effect produced by coincubation with saturating concentrations of HS was no longer apparent if the cells had been pretreated with heparitinase. These findings distinguish quantitatively between neurite growth on laminin and on laminin-HSPG complexes, and suggest that some neuronal cell surface and released HSPGs are involved in neurite growth by virtue of non-covalent interactions with glycosaminoglycan binding domains of laminin.