Functional joint regeneration is achieved using reintegration mechanism in Xenopus laevis.

Rio Tsutsumi,Shigehito Yamada,Kiyokazu Agata

doi:10.1002/reg2.49

Abstract

A functional joint requires integration of multiple tissues: the apposing skeletal elements should form an interlocking structure, and muscles should insert into skeletal tissues via tendons across the joint. Whereas newts can regenerate functional joints after amputation, Xenopus laevis regenerates a cartilaginous rod without joints, a “spike.” Previously we reported that the reintegration mechanism between the remaining and regenerated tissues has a significant effect on regenerating joint morphogenesis during elbow joint regeneration in newt. Based on this insight into the importance of reintegration, we amputated frogs’ limbs at the elbow joint and found that frogs could regenerate a functional elbow joint between the remaining tissues and regenerated spike. During regeneration, the regenerating cartilage was partially connected to the remaining articular cartilage to reform the interlocking structure of the elbow joint at the proximal end of the spike. Furthermore, the muscles of the remaining part inserted into the regenerated spike cartilage via tendons. This study might open up an avenue for analyzing molecular and cellular mechanisms of joint regeneration using Xenopus.

Highlights

Limb regenerative ability varies among vertebrates (Wallace1981; Agata & Inoue 2012; Seifert et al 2012)
To investigate the effect of the remaining tissues on the morphogenesis of regenerated tissues in frog, we amputated frogs’ forelimbs slightly distal to the elbow joint and removed the residual amputated radio-ulna from the stylopod (Fig. 1). After this amputation at the elbow joint, 89.5% (17/19) of frogs regenerated a spike on the stump, a rate comparable to that in frogs whose limb was amputated at the stylopod or the zeugopod level (Tassava 2004)
The distal side of the original elbow joint was completely eliminated by the surgery, frogs showed bending and stretching motions of their elbow joint in the regenerated forelimb (Fig. 2A−C; Movie S1)

Summary

Introduction

Limb regenerative ability varies among vertebrates (Wallace1981; Agata & Inoue 2012; Seifert et al 2012). Many urodele amphibians (newts and salamanders) are able to regenerate the complete structure of their limbs after amputation throughout their lives. In anuran amphibians (frogs and toads), perfect limb regeneration is only observed during the larval stage, and they become unable to regenerate the complete structure of the limb after metamorphosis Adult Xenopus laevis, for example, regenerate a hypomorphic single cartilage rod called a spike, without joints, muscles, mineralized bones, tendons, or ligaments, after amputation at any level along the limb axis (Endo et al 2000; Tassava 2004; Satoh et al 2005a, 2006; Suzuki et al 2006). Adult frog limb regeneration is regarded as an intermediate model between urodele amphibian and mammalian limb regeneration (Seifert et al 2012)

Methods

Results

Conclusion