Neuromuscular specificity following cross-stage hindlimb transplantation

Abstract

In order to determine whether spinal motoneurons can regenerate to their proper targets at stages beyond those when such specificity is typically expressed, autologous and homologous (same- and cross-stage) hindlimb transplantations were performed using bullfrog tadpoles ( Rana catesbeiana). Neuromuscular specificity was assessed by applying horseradish peroxidase to the ventral thigh of the transplanted hindlimb and mapping the locations of retrogradely labeled motoneurons. Previously, we found that the hindlimb was reinnervated normally in young tadpoles whose motor nerves were transected. However, motor axons in older animals showed no evidence of target specificity when reinnervating the tadpole's own limb. In the present study, innervation was normal in young tadpoles whose hindlimb was removed and replaced in its original position. Axons of older hosts innervating hindlimbs transplanted from young tadpoles distinguished flexor from extensor limb regions, but failed to distinguish thigh from shank, demonstrating that axons in older animals can respond to at least some guidance cues. The lack of specificity along the proximal-distal axis appears to be a consequence of homologous transplantation since limbs transplanted from one young tadpole to another showed the same loss of proximal-distal specificity. One possibility is that the shank degenerates when the hindlimb is transplanted to a different host. Shank motoneurons, lacking their proper target, may then innervate the thigh. However, shank motoneurons did not innervate the thigh when motor nerve transection was combined with amputation of the hindlimb just above the presumptive knee. These results show: (i) motoneurons in older tadpoles have the capability for greater regenerative specificity than is normally expressed and (ii) regenerative specificity in the proximal-distal dimension depends upon matching of motoneurons with particular targets rather than upon a hierarchy of preferred affinities.