RNA-Sequencing of Xenopus tail regeneration using small molecule antagonists of Hif1a or Wnt signaling

Regeneration of complex tissues is initiated by an injury-induced stress response, eventually leading to activation of developmental signaling pathways, such as Wnt signaling. How early injury cues are interpreted and coupled to activation of these developmental signals and their targets is not well understood. Here, we show that Hif1a, a stress induced transcription factor, is required for tail regeneration in Xenopus tropicalis. We find that Hif1a is required for regeneration of differentiated axial tissues, including axons and muscle. Using RNA-sequencing, we find that Hif1a and Wnt converge on a broad set of genes required for posterior specification and differentiation, including the posterior hox genes. We further show that Hif1a is required for transcription via a Wnt-responsive element, a function that is conserved in both regeneration and early neural patterning. Our findings indicate a regulatory role for Hif1a in Wnt mediated gene expression across multiple tissue contexts. Overall design: Tail tissue proximal to the amputation site (for 0hpa) or regenerating tissue (24hpa following treatment with DMSO, 2-methoxyestradiol, echinomycin, or IWR-1) was collected for RNA extraction. Paired end sequencing was performed on prepared libraries via an Illumina HiSeq instrument.