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Hydra  is a unique model for studying the mechanisms underlying stem cell biology. The activity of the three stem cell lineages structuring its body constantly replenishes mature cells lost due to normal tissue turnover. By a poorly understood mechanism, stem cells are maintained through self-renewal while concomitantly producing differentiated progeny. In vertebrates, one of many genes that participate in regulating stem cell homeostasis is the protooncogene  c-myc , which has been recently identified also in  Hydra , and found expressed in the interstitial stem cell lineage. In the present paper, by developing a novel strategy of RNA interference-mediated gene silencing (RNAi) based on an enhanced uptake of small interfering RNAi (siRNA), we provide molecular and biological evidence for an unexpected function of the  Hydra myc  gene  (Hymyc1)  in the homeostasis of the interstitial stem cell lineage. We found that  Hymyc1  inhibition impairs the balance between stem cell self renewal/differentiation, as shown by the accumulation of stem cell intermediate and terminal differentiation products in genetically interfered animals. The identical phenotype induced by the 10058-F4 inhibitor, a disruptor of c-Myc/Max dimerization, demonstrates the specificity of the RNAi approach. We show the kinetic and the reversible feature of  Hymyc1  RNAi, together with the effects displayed on regenerating animals. Our results show the involvement of  Hymyc1  in the control of interstitial stem cell dynamics, provide new clues to decipher the molecular control of the cell and tissue plasticity in  Hydra , and also provide further insights into the complex  myc  network in higher organisms. The ability of  Hydra  cells to uptake double stranded RNA and to trigger a RNAi response lays the foundations of a comprehensive analysis of the RNAi response in  Hydra  allowing us to track back in the evolution and the origin of this process.

Hymyc1 Downregulation Promotes Stem Cell Proliferation in Hydra vulgaris