Cell cycle and histological effects of reinnervation in denervated forelimb stumps of larval Ambystoma

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Abstract
Although nerves are a requirement for regeneration of a salamander limb, denervation of an amputated larval Ambystoma forelimb does not permanently prevent the limb from regenerating. When nerves grow back into the distal stump in sufficient numbers, regeneration occurs. In this study we examined histological and cell cycle events in denervated limb stumps of Ambystoma as they became reinnervated and began to regenerate. Prior to reinnervation and regeneration, dedifferentiated cells are present in the distal stump but are rarely synthesizing DNA or undergoing mitosis and are densely packed. About 2 days after the distal tip of the stump becomes reinnervated, the limb tip appears to “expand” as cells become less densely packed, and cell cycle activity begins. Mitotic index (MI) and labeling index (LI) increase slowly compared to MI and LI in control amputated limbs. Evidence for the localization of cycling activity is presented. In reinnervated stumps, the increase in LI precedes the increase in MI, suggesting that most of the cells are initiating cycling from the G1 phase.