A staging system has been devised for normal regeneration from the upper arm in the mature axolotl. It consists of seven externally definable stages: (1) Wound healing (WH): (2) Dedifferentiation (DD); (3) Early bud (EB); (4) Medium bud (MB); (5) Late bud (LB); (6) Palette (Pal), and (7) Digital outgrowth (DO). Serial histological sections of 38 regenerating limbs were used to correlate gross stages with microscopic events in the regenerative process.
A comparison between the surface features of embryonic limb development and limb regeneration was made in the axolotl. Scanning electron microscopy revealed an overall similarity between embryonic and regenerating limbs. A notable feature was the lack of a morphologically discrete apical epidermal specialization on the surface of any of the limbs. Histological preparations revealed no thickening of the apical epidermis in embryonic limbs. There is a definite thickening of the apical epidermis in regenerating limbs during the stages of dedifferentiation and the early blastemal phases, but the thickening projects into the underlying tissues rather than altering the smooth surface contours of the blastema. Minor differences in epidermal specializations (ciliated cells scattered over the entire body and very irregularly shaped cells with knob-like projections at the tips of digits in embryos) may be attributable to differences in overall activity of the animals at various stages in their life history.
Although capable of initiating early regenerative responses, axolotl forelimb stumps which are composed of double-half limb tissues fail to undergo the events that normally lead to the replacement of missing parts. In the present study, the posterior halves of right forelimbs were exchanged with the anterior halves of left forelimbs, or the dorsal halves of right forelimbs were exchanged with the ventral halves of left forelimbs. Forelimbs were amputated through the graft region 30 days after grafting. Limb stumps bearing double-dorsal, double-ventral or double-posterior tissues either produced hypomorphic regenerates or failed to form any externally visible outgrowth. When the limb stump bore double-anterior tissues, no externally visible structures were formed. Normal and multiple regenerates were never formed by double-half limbs. These results are discussed in terms of the polar coordinate model and suggest that the regeneration blastema requires a complete circumference of positional values in order to complete distal transformation.
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