Axolotl (Ambystoma mexicanum) Limb and Tail Amputation

Abstract

INTRODUCTIONAppendage regeneration is a well-known and well-studied trait in salamanders including the axolotl (Ambystoma mexicanum). Depending on the size of the animal, functional regeneration of the limb or tail can occur within 3 wk (in 2-cm-long animals) to several months (in >10-cm-long animals). After appendage amputation, epithelial wound healing is followed by formation of a progenitor cell zone called the blastema that first undergoes expansion and then morphogenesis and differentiation to replace the appendage. To obtain reproducible, morphologically perfect regeneration, it is important to cut the appendage so that the wound site is evenly open over its surface. This allows cells from the basal layer of the surrounding epidermis to crawl over the cut end. This protocol describes limb amputation in 3- to 5-cm and 5- to 8-cm axolotls, as well as tail amputation in 3- to 8-cm axolotls.