Abstract
Axolotls have amazing ability to regenerate their lost limbs. Our previous works showed that after amputation the remnant muscle ends remained at their original location whilst sending satellite cells into the regenerating parts to develop into early muscle fibers in the late differentiation stage. The parental and the newly formed muscle fibers were not connected until very late stage. The present study used non-invasive diffusion tensor imaging (DTI) to monitor weekly axolotl upper arm muscles after amputation of their upper arms. DTI tractography showed that the regenerating muscle fibers became visible at 9-wpa (weeks post amputation), but a gap was observed between the regenerating and parental muscles. The gap was filled at 10-wpa, indicating reconnection of the fibers of both muscles. This was confirmed by histology. The DTI results indicate that 23% of the muscle fibers were reconnected at 10-wpa. In conclusion, DTI can be used to visualize axolotls’ skeletal muscles and the results of muscle reconnection were in accordance with our previous findings. This non-invasive technique will allow researchers to identify the timeframe in which muscle fiber reconnection takes place and thus enable the study of the mechanisms underlying this reconnection.
Highlights
Axolotls are well known for their amazing abilities to regenerate lost limbs after amputation [1]
In our previous work [3], we found that the regeneration of axolotl limb muscles after limb amputation does not progress piece by piece from the remnant muscle ends
The aim of the present study is to show the feasibility of diffusion tensor imaging (DTI) to monitor muscle growth in axolotls after amputation, and to provide in vivo evidence to support our previous observations that the reconnection between regenerating and parental muscles occurs during late differentiation stage
Summary
Axolotls are well known for their amazing abilities to regenerate lost limbs after amputation [1]. Amputation of the limbs results in the formation of blastemas in the stump ends, which contains undifferentiated cells capable of growing and developing into new limbs as they were before amputation [2]. In our previous work [3], we found that the regeneration of axolotl limb muscles after limb amputation does not progress piece by piece from the remnant muscle ends. Satellite cells from the remnant muscle ends migrate into the blastemas since the early or mid-bud stage, and further migrate in tracts into the regenerating limbs during the early and late differentiation stages.
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