The effects of combining chondroitinase ABC and NEP1-40 on the corticospinal axon growth in organotypic co-cultures

Abstract The area surrounding the injured spinal cord is a non-permissive milieu for axonal growth due to the inhibitory factors, especially chondroitin sulfate proteoglycan (CSPG) and Nogo. Recent studies have reported that chondroitinase ABC (ChABC) or Nogo-66(1-40) antagonist peptide (NEP1-40) promote axonal growth after spinal cord injury. But no study has addressed the effects on spinal cord injury of combining ChABC and NEP1-40. Previously, we described an organotypic co-culture system using the brain cortex and spinal cord from neonatal rats. In this study, we examined whether the combination of ChABC and NEP1-40 creates an action that promotes corticospinal axon growth in organotypic co-cultures. Organotypic co-cultures of brain and spinal cord were prepared from rats, and ChABC or NEP1-40 was delivered to them. To examine the effects of this combination these two drugs were applied together. We counted the number of labeled axons with DiI and assessed the immunoreactivity of CSPG and Nogo. Axonal growth was enhanced by infusing ChABC or NEP1-40 compared with that in the control group, whereas synergistic effects of combined administration of ChABC and NEP1-40 on axonal growth were not observed. There is a possibility that ChABC and NEP1-40 affect the same intracellular pathways and have no synergistic influence on axonal growth.