Regeneration of Adult Rat Sensory Axons into Intraspinal Nerve Grafts: Promoting Effects of Conditioning Lesion and Graft Predegeneration

Abstract

The effect of intraspinally implanted predegenerated peripheral nerve grafts and/or conditioning lesions on the regenerative capacity of central ascending sensory axons was investigated in the adult rat. Regenerating sensory fibers were analyzed after their transganglionic labeling with cholera toxin B subunit, usually 1 month after implantation. A conditioning lesion (transection of the tibial and peroneal nerve) caused a fivefold increase in the number of sensory fibers within the fresh graft when applied on the day of grafting and a sevenfold increase when applied 1 week before. In the latter case, a small portion of the fibers (10%) had reached the rostral end of the nerve graft. In the absence of a conditioning lesion, the number of fibers regenerating into a predegenerated nerve graft (collected from the distal part of the peroneal nerve that had been axotomized 1 week earlier) was similar to that found in a fresh graft. However, predegenerated grafts received three and five times more fibers than a fresh graft when conditioning lesions were applied on or 1 week before the day of grafting. With the combination of a predegenerated graft and a 1-week conditioning, most (>90%) of the regenerating fibers had reached the rostral graft-host border. In animals with a fresh graft, a portion of the axotomized fibers formed terminal club-like structures. Much fewer fibers displayed such clubs in animals with a predegenerated graft or a conditioning lesion, suggesting a preventive action of either treatment. A time-course study with the combined treatments showed that regenerating sensory fibers had already entered the graft after 3 days. Between 1 and 2 weeks, a maximum number of fibers had reached the rostral end of the nerve graft. However, after 2 months, the number of fibers was decreased, i.e., the initial advantage of predegeneration had diminished. The current results demonstrate the necessity of a conditioning lesion for successful regeneration of central sensory fibers, possibly resulting from an earlier induction of the neuronal growth response which allows the axon to enter the graft before the formation of a graft-host barrier. The predegeneration of the nerve graft augments the growth response of the axotomized central sensory fibers, probably by providing a more supportive terrain and/or enhancement of the neuronal response. The presence of a large number of fibers at the rostral graft-host border now provides the opportunity to investigate the effects of neurotrophic factors on the regenerative capacity of the ascending rat sensory fibers into the denervated spinal cord in vivo.