Chondroitin sulfate-binding peptides block chondroitin 6-sulfate inhibition of cortical neurite growth

Chondroitin sulfate (CS) expression is increased in the glial scar following spinal cord injury demonstrating the importance understanding the role of CS in the central nervous system (CNS). There have been conflicting studies on the effects of the most abundant types of CS, chondroitin 4-sulfate (C4S) and chondroitin 6-sulfate (C6S), found in the CNS. In this study, the effects of C4S and C6S on rat embryonic day 18 cortical neurons were investigated. C4S had no effect on neuron behavior whereas C6S inhibited neurite outgrowth at higher concentrations (>10 μg/ml). Two C6S-binding peptides (C6S-1 and C6S-2) were tested for their ability to block the inhibitory activity of C6S on neurite outgrowth. Neurons cultured with C6S and C6S-binding peptide at higher peptide concentrations had neurite lengths similar to neurons cultured without C6S. Therefore, the C6S-binding peptides were effective at blocking the inhibitory activity of C6S. The C6S-1 peptide had a higher binding affinity than the C6S-2 peptide and was consequently more effective at blocking C6S inhibition of neurite growth. To date, this is the first study to employ an alternative strategy from enzymatic digestion of CS chains to increase neurite outgrowth. These studies warrant further investigation of the use of C6S-binding peptides to increase nerve regeneration following spinal cord injury.