RRC ID 77468
Author 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.
Title The shh limb enhancer is activated in patterned limb regeneration but not in hypomorphic limb regeneration in Xenopus laevis.
Journal Dev Biol
Abstract 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.
Volume 500
Pages 22-30
Published 2023-8-1
DOI 10.1016/j.ydbio.2023.05.009
PII S0012-1606(23)00093-3
PMID 37247832
MeSH Animals Animals, Genetically Modified Epigenesis, Genetic* Extremities* / physiology Transcription Factors / genetics Xenopus laevis / genetics
IF 2.896
DNA material MFCS1-luciferace (RDB19989) MFCS1-pshh-luciferace (RDB19990) pCMV-Hoxd13 (RDB19991) pCMV-Fgf2 (RDB19992) pCMV-Fgf4 (RDB19993) pCMV-Fgf8 (RDB19994)