Sources of trophic factors that induce limb regeneration and prevent amputation-induced neuronal death

Segments of 2-, 4-, 6- and 8-day neural tube, or of 15-day peripheral nerve were implanted longitudinally into limb stumps of 4-day chick embryos whose right-wing buds were amputated at the future elbow region. Stumps of amputated limbs (ALs) implanted with 7-day heart or without implant served as controls. Effects of progressively older neural tube implants (NTIs) upon ALs and host spinal cord neurons were analyzed by area measurements of the peripheral limb field (PLF) and NTI and by cell counts of the host lateral motor column (LMC). Nine days postamputation, 2- and 4-day NTIs contained many neurons and induced epimorphic regeneration in more than one-fourth of the embryos. Six-day NTIs contained few neurons and induced only tissue regeneration. Eight-day NTIs and peripheral nerve containing only non-neuronal cells were as ineffective as controls in stimulating regeneration, although peripheral nerve did cause a significant increase in the peripheral field. The NTIs of all ages and implants of peripheral nerve were equally effective in protecting LMC neurons from amputation-induced cell death in the host spinal cord. The results may indicate that neurons of the implant induce limb regeneration and non-neuronal cells of the implant protect against LMC neuronal death.