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Dev Biol 2023 May 27;500:22-30. doi: 10.1016/j.ydbio.2023.05.009.
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The shh limb enhancer is activated in patterned limb regeneration but not in hypomorphic limb regeneration in Xenopus laevis.

Tada R , Higashidate T , Amano T , Ishikawa S , Yokoyama C , Kobari S , Nara S , Ishida K , Kawaguchi A , Ochi H , Ogino H , Yakushiji-Kaminatsui N , Sakamoto J , Kamei Y , Tamura K , Yokoyama H .

Xenopus young tadpoles regenerate a limb with the anteroposterior (AP) pattern, but metamorphosed froglets regenerate a hypomorphic limb after amputation. The key gene for AP patterning, shh, is expressed in a regenerating limb of the tadpole but not in that of the froglet. Genomic DNA in the shh limb-specific enhancer, MFCS1 (ZRS), is hypermethylated in froglets but hypomethylated in tadpoles: shh expression may be controlled by epigenetic regulation of MFCS1. Is MFCS1 specifically activated for regenerating the AP-patterned limb? We generated transgenic Xenopus laevis lines that visualize the MFCS1 enhancer activity with a GFP reporter. The transgenic tadpoles showed GFP expression in hoxd13-and shh-expressing domains of developing and regenerating limbs, whereas the froglets showed no GFP expression in the regenerating limbs despite having hoxd13 expression. Genome sequence analysis and co-transfection assays using cultured cells revealed that Hoxd13 can activate Xenopus MFCS1. These results suggest that MFCS1 activation correlates with regeneration of AP-patterned limbs and that re-activation of epigenetically inactivated MFCS1 would be crucial to confer the ability to non-regenerative animals for regenerating a properly patterned limb.

PubMed ID: 37247832
Article link: Dev Biol

Species referenced: Xenopus laevis
Genes referenced: dusp6 fgf2 fgf4 fgf8 hoxd13 shh
GO keywords: regeneration [+]

Phenotypes: Xla + 180degree rotation (Fig. 2 DF)

Article Images: [+] show captions