INFLUENCE OF N-LINKED OLIGOSACCHARIDES ON THE PROCESSING AND NEURITE-PROMOTING ACTIVITY OF PROTEOGLYCANS RELEASED BY NEURONS INVITRO

Inhibitors of enzymes involved in processing of N-linked oligosaccharides were used to examine biosynthesis and the neurite-promoting activity of proteoglycans produced by and released from dissociated chick embryo spinal cord neurons in vitro. In the cell compartment and in conditioned medium both castanospermine and swainsonine inhibited H-3-glucosamine incorporation into glycoprotein but only castanospermine reduced H-3-glucosamine incorporation into heparan sulphate and chondroitin sulphate proteoglycans. All of the neurite-promoting activity of neuron-conditioned medium that complexed to laminin was associated with heparan sulphate proteoglycans as determined by heparitinase digestion. Neuron-conditioned medium prepared in the presence of castanospermine displayed a 38 +/- 6% (mean +/- SD) reduction in H-3-glucosamine incorporation into heparan sulphate proteoglycans and a 30 +/- 5% reduction in substrate-attached neurite-promoting activity compared to control conditioned medium and to conditioned medium prepared in the presence of swainsonine. When neurons were coincubated with castanospermine, neurite growth on a laminin substrate was 50 +/- 10% of control growth or growth in the presence of swainsonine. However, when neuron-conditioned medium was used to pretreat the laminin substrate the inhibitory effect on neurite growth produced by castanospermine coincubation was reversed. Influences on neuronal processing of N-linked oligosaccharides alter neurite growth directly and also alter the neurite-promoting activity of neuron-conditioned medium by inhibiting the synthesis of heparan sulphate proteoglycans. These studies provide further evidence for an autocrine role for heparan sulphate proteoglycans in neurite growth.