Growth associated protein 43 and neurofilament immunolabeling in the transected lumbar spinal cord of lizard indicates limited axonal regeneration

Previous cytological studies on the transected lumbar spinal cord of lizards have shown the presence of differentiating glial cells, few neurons and axons in the bridge region between the proximal and distal stumps of the spinal cord in some cases. A limited number of axons (20-50) can cross the bridge and re-connect the caudal stump of the spinal cord with small neurons located in the rostral stump of the spinal cord. This axonal regeneration appears to be related to the recovery of hind-limb movements after initial paralysis. The present study extends previous studies and shows that after transection of the lumbar spinal cord in lizards, a glial-connective tissue bridge that reconnects the rostral and caudal stumps of the interrupted spinal cord is formed at 11-34 days post-injury. Following an initial paralysis some recovery of hindlimb movements occurs within 1-3 months post-injury. Immunohistochemical and ultrastructural analysis for a growth associated protein 43 (GAP-43) of 48-50 kDa shows that sparse GAP-43 positive axons are present in the proximal stump of the spinal cord but their number decreased in the bridge at 11-34 days post-transection. Few immunolabeled axons with a neurofilament protein of 200-220 kDa were seen in the bridge at 11-22 days post-transection but their number increased at 34 days and 3 months post-amputation in lizards that have recovered some hindlimb movements. Numerous neurons in the rostral and caudal stumps of the spinal cord were also labeled for GAP43, a cytoplasmic protein that is trans-located into their axonal growth cones. This indicates that GAP-43 biosynthesis is related to axonal regeneration and sprouting from neurons that were damaged by the transection. Taken together, previous studies that utilized tract-tracing technique to label the present observations confirm that a limited axonal re-connection of the transected spinal cord occurs 1-3 months post-injury in lizards. The few regenerating-sprouting axons within the bridge reconnect the caudal with the rostral stumps of the spinal cord, and likely contribute to activate the neural circuits that sustain the limited but important recovery of hind-limb movements after initial paralysis. The surgical procedures utilized in the study followed the regulations on animal care and experimental procedures under the Italian Guidelines (art. 5, DL 116/92).