Functional Role of Myc Transcript as Active Player in the Canonical Wnt Pathway During Intestinal Regeneration

Abstract

Molecular approaches have been widely used to explore the role of genes associated with regenerative processes in different animal models. Our own studies in the sea cucumber Holothuria glaberrima have identified several genes that appear to play a role in the regeneration of the digestive system. We have focused on two of these genes, Wnt and Myc, whose expression has been localized to the intestinal rudiment during regeneration. Myc and Wnt are interrelated, as the former is known to serve as an active player in the canonical Wnt signaling pathway. We now have quantified the levels of myc transcript at different stages of regeneration. In addition, we designed Dicer‐substrate short interfering RNAs (DsiRNAs)‐targeting Myc transcripts. DsiRNAs‐targeting Myc are electroporated in regenerating intestinal tissue explants of H. glaberrima to evaluate their effects on cell proliferation and cell dedifferentiation. Quantification of myc transcripts shows a decrease following electroporation of specific DsiRNAs when compared to explants electroporated with control DsiRNAs. Moreover, a decrease in cell proliferation is observed following the reduction in myc transcript, suggesting that Myc has a functional role on cell proliferation during the intestinal regeneration process. Instead, cell dedifferentiation does not change with the interference of the expression of Myc, suggesting that Myc does not play an important functional role on cell dedifferentiation during the intestinal regeneration process. In summary, this project not only presents data on Wnt‐Myc interactions, but presents three important technical advances: (1) the use of intestinal tissue explants, (2) the implementation of the electroporation as a transfection method and (3) the use of DsiRNAs to manipulate the gene expression in H. glaberrima explants.Support or Funding InformationFunded by NIH R15GM124595, NSF IOS‐1252679 and the University of Puerto RicoThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.