Unknown,Transcriptomics,Genomics,Proteomics

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Selective amputation of the pharynx identifies a FoxA-dependent regeneration program in planaria

ABSTRACT: Planarian flatworms can regenerate every organ after amputation. Adult pluripotent stem cells drive the ability to regenerate, but how injury activates these cells and directs them into the appropriate lineages is not understood. To study the regeneration response in a simplified context, we selectively induced the ejection of the planarian pharynx by briefly exposing animals to sodium azide followed by microarray analysis. Tissue surrounding the wound site was examined at 6, 12, 18, 24, 48, and 72 hours post amputation by comparison to tissue isolated immediately after amputation (time zero). Stem cells are required for the regeneration process, and can be eradicated by exposure to radiation. A parallel time course was also performed on worms that had been irradiated with 10k rads of gamma irradiation. Two time courses consisting of irradiated or unirradiated worms following chemical amputation of the pharynx. Tissue samples taken at 6, 12, 18, 24, 48 and 72 hours are each compared to a reference sample at time zero. The experiment was performed in triplicate for a total of 42 samples on 36 arrays.

ORGANISM(S): Schmidtea mediterranea

SUBMITTER: Chris Seidel

PROVIDER: E-GEOD-56181 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Variability of regenerative potential among animals has long perplexed biologists. Based on their amazing regenerative abilities, planarians have become important models for understanding the molecular basis of regeneration; however, planarian species with limited regenerative abilities are also found. Despite the importance of understanding the differences between closely related, regenerating and non-regenerating organisms, few studies have focused on the evolutionary loss of regeneration, and the molecular mechanisms leading to such regenerative loss remain obscure. Here we examine Procotyla fluviatilis, a planarian with restricted ability to replace missing tissues, utilizing next-generation sequencing to define the gene expression programs active in regeneration-permissive and regeneration-deficient tissues. We found that Wnt signaling is aberrantly activated in regeneration-deficient tissues. Remarkably, down-regulation of canonical Wnt signaling in regeneration-deficient regions restores regenerative abilities: blastemas form and new heads regenerate in tissues that normally never regenerate. This work reveals that manipulating a single signaling pathway can reverse the evolutionary loss of regenerative potential. RNA-seq experiments to identify gene expression changes following amputation in body regions with variable regenerative potential. Adult Procotyla fluviatilis were amputated at sites either anterior or posterior to the pharynx. After 24 hours post-amputation, tissues near the amputation site were excised and RNA was extracted. Similar tissues were excised from uncut control animals. Samples were processed for RNA-seq using Illumina procedures. We generated a de novo P. fluviatilis transcriptome and used RNA sequencing (RNA-seq) to characterize transcripts from excised tissue fragments in Reg+ and Reg- body regions 24 hours post-amputation. We performed parallel analyses on tissues excised from intact animals at identical body regions to account for regional differences in transcripts, thereby identifying changes resulting from amputation. Samples A1-A3 = Regeneration-proficient (Reg+) tissue excision 24 hours after amputation. Samples B1-B3 = Tissue excision from regeneration-proficient (Reg+) region but not amputated. Samples C1-C3 = Tissue excision from regeneration-deficient (Reg-) tissues 24 hours after amputation. Samples D1, D3-D4 = Tissue excision from regeneration-deficient (Reg-) region that was not amputated.

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Selective amputation of the pharynx identifies a FoxA-dependent regeneration program in planaria

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