Evidence for new growth and regeneration of cut axons in developmental plasticity of the rubrospinal tract in the north american opossum

We have shown previously that rubral axons can grow around a lesion of their spinal pathway in the developing opossum and that a critical period exists for that plasticity (Martin and Xu, Dev Brain Res 39:303, 1988). Since most rubrospinal neurons degenerate after axotomy during the critical period, we have proposed that plasticity results primarily from growth of late arriving axons around the lesion rather than regeneration of cut axons (Xu and Martin, J Comp Neurol 279:368, 1989). In the present study, we used a double‐labeling paradigm to test that hypothesis. Four groups of pouch young opossums received bilateral or unilateral injections of Fast Blue (FB) into the caudal thoracic or rostral lumbar cord (T12–L2) at different ages in order to label rubrospinal neurons. Three or 4 days later, the rubrospinal tract was transected unilaterally, four to five segments rostral to the injection(s). If the injection was unilateral, the lesion was made ipsilateral to it. The animals were maintained for about 1 month before a second marker, Diamidino Yellow (DY), was injected, usually bilaterally, between the FB injection(s) and the lesion. The animals were maintained for about 5 days before sacrifice and sections through the red nucleus and spinal cord were examined with a fluorescence microscope. During the critical period for plasticity, only a few rubral neurons contralateral to the lesion were labeled by FB alone, supporting our previous contention that most axotomized neurons degenerate. In contrast, many neurons were labeled by DY alone, indicating that their axons were not present in the caudal cord at the time of the FB injection and that they grew around the lesion during the 1 month survival to incorporate DY. A few double‐labeled neurons were also found. One interpretation of such neurons is that they survived axotomy, as evidenced by the presence of FB, and supported axons which grew around the lesion to take up DY. Another interpretation is that they supported late growing axons which incorporated residual FB as well as DY. In order to choose between these alternatives, a similar double‐labeling paradigm was carried out, but with removal of FB at the time of the lesion. Since a few neurons were still double labeled, we conclude that regeneration of cut axons also contributed to rubrospinal plasticity. Our results support our previous suggestion that developmental plasticity of the rubrospinal tract results primarily from growth of late arriving axons around the lesion, but they also suggest that regeneration of cut axons occurs.