Morphogenetic properties of the skin in axolotl limb regeneration.

A study has been made of the morphogenetic properties of anterior and posterior skin from the lower forelimb of the axolotl. The basic experiment consisted of a graft of a half cuff of skin from a donor to a host limb followed by a 2-week healing period, amputation through the graft, and a study of the resulting regenerate. Limbs with double posterior skin formed double posterior regenerates and, in contrast, limbs with double anterior skin formed normal or slightly hypomorphic regenerates. Posterior skin from post-metamorphic animals had a similar but weaker effect to that from ordinary axolotls. Immunological rejection of allografts could be completely avoided if the donor limb was transplanted to the flank of the host when both were at the stage of tail-bud embryos, and the skin graft was later carried out between the supernumerary limb and one of the host limbs. This technique was used to show that immunological rejection does not affect the formation of duplicates from the limbs with double posterior skin, and to facilitate the studies of the cellular provenance of the regenerate. The cellular composition of duplicate regenerates was studied by using both triploid donors and triploid hosts. It was shown that the posterior side of the duplications consisted wholly of host tissue and the anterior side consisted of mixed donor and host tissue. Formation of the duplicated regenerate therefore seems to involve positional reprogramming of both donor and host tissues together with metaplasia of the donor tissue. It was not possible to inhibit the duplication-inducing property of posterior skin by treatment with a variety of enzymes. A model based on the serial threshold theory of regeneration is advanced to explain the results. This model successfully accounts for the observed non-equivalence of anterior and posterior skin, and also explains the different regeneration behaviour of anterior and posterior half limbs, the limited regeneration of double anterior limbs, and the pattern expansion and contraction shown by regenerates from double posterior limbs.