Functional Analyses of Myc and β‐catenin during Echinoderm Intestinal Regeneration using RNAi

Abstract

Molecular approaches have been widely used to understand and characterize regenerative processes in different animal systems. Nevertheless, characterizing the genetic component of echinoderms using RNAi techniques represent a tremendous challenge. We have now designed and implemented the use of Dicer‐substrate RNAi technology (DsiRNA) to knockdown the expression of Myc and β‐catenin transcripts on tissue explants of the sea cucumber Holothuria glaberrima. Quantification of Myc and β‐catenin transcripts showed a decrease following electroporation of specific DsiRNAs when compared to explants electroporated with control DsiRNA. Explants were then assayed for the effects on cell proliferation, cell dedifferentiation and apoptosis. No effect on cellular dedifferentiation was observed. In contrast, the number of apoptotic cells was significantly different in explants electroporated with DsiRNAs‐targeting Myc and β‐catenin when compared to control DsiRNA. Finally, a decrease in cell proliferation was observed following the reduction of Myc transcript, but not following the reduction of β‐catenin. These results suggest that Myc has a significant role on cell proliferation and apoptosis, but not on cell dedifferentiation during the regeneration of the intestine. Similar to Myc, β‐catenin has a significant role on apoptosis, but not on cell proliferation and cell dedifferentiation. Moreover, our results show that DsiRNAs and intestinal tissue explants are a valuable tool to manipulate and explore the role of genes of interest in echinoderms.Support or Funding InformationFunded by NIH R15GM124595, NSF IOS‐1252679 and the University of Puerto Rico.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.