Cnidarians possess remarkable powers of regeneration, but the cellular and molecular mechanisms underlying this capability are unclear. Studying the hydrozoan Hydractinia echinata we show that a burst of stem cell proliferation occurs following decapitation, forming a blastema at the oral pole within 24 hr. This process is necessary for head regeneration. Knocking down Piwi1, Vasa, Pl10 or Ncol1 expressed by blastema cells inhibited regeneration but not blastema formation. EdU pulse-chase experiments and in vivo tracking of individual transgenic Piwi1+ stem cells showed that the cellular source for blastema formation is migration of stem cells from a remote area. Surprisingly, no blastema developed at the aboral pole after stolon removal. Instead, polyps transformed into stolons and then budded polyps. Hence, distinct mechanisms act to regenerate different body parts in Hydractinia. This model, where stem cell behavior can be monitored in vivo at single cell resolution, offers new insights for regenerative biology.DOI: http://dx.doi.org/10.7554/eLife.05506.001

Clonal animals do not sequester a germ line during embryogenesis. Instead, they have adult stem cells that contribute to somatic tissues or gametes. How germ fate is induced in these animals, and whether this process is related to bilaterian embryonic germline induction, is unknown. We show that transcription factor AP2 (Tfap2), a regulator of mammalian germ lines, acts to commit adult stem cells, known as i-cells, to the germ cell fate in the clonal cnidarian Hydractinia symbiolongicarpus. Tfap2 mutants lacked germ cells and gonads. Transplanted wild-type cells rescued gonad development but not germ cell induction in Tfap2 mutants. Forced expression of Tfap2 in i-cells converted them to germ cells. Therefore, Tfap2 is a regulator of germ cell commitment across germ line–sequestering and germ line–nonsequestering animals.

SUMMARYWe studied the role of Wnt signaling in axis formation during metamorphosis and regeneration in the cnidarian Hydractinia. Activation of Wnt downstream events during metamorphosis resulted in a complete oralization of the animals and repression of aboral structures (i.e. stolons). The expression of Wnt3, Tcf and Brachyury was upregulated and became ubiquitous. Rescue experiments using Tcf RNAi resulted in normal metamorphosis and quantitatively normal Wnt3 and Brachyury expression. Isolated, decapitated polyps regenerated only heads but no stolons. Activation of Wnt downstream targets in regenerating animals resulted in oralization of the polyps. Knocking down Tcf or Wnt3 by RNAi inhibited head regeneration and resulted in complex phenotypes that included ectopic aboral structures. Multiple heads then grew when the RNAi effect had dissipated. Our results provide functional evidence that Wnt promotes head formation but represses the formation of stolons, whereas downregulation of Wnt promotes stolons and represses head formation.

Adult colonies of the reef-building coral St lophora pistillata discriminate precisely between ' self ' and ' nonself ' attributes, and respond selectively against specific allogeneic challenges. We studied the ontogeny of these allospecific responses on newly settled polyps by establishing allogeneic contacts within groups of 2-6 siblings or non-related offspring. Interactions were observed for up to 8 months. Three types of response, depending on the age of the interacting partners, were documented. The first was tissue fusion and the formation of a stable chimera, observed in partners less than 2 months old. The second was observed in contacts of partners 2-4 months old. It started with tissue fusion and transitory chimera since separation of the chimera partners or polyp death resulted when the oldest partner in the chimera reached the age of 4 months. The third type was the regular histoincompatibility response, as documented in allogeneic interactions of adult colonies, recorded here in all encounters with S. pistillata partners over 4 months old. Maturation of allorecognition in this species was therefore achieved through three time-dependent stages, 4 months following metamorphosis. Combinations of siblings or genetically unrelated partners did not affect the results. We propose that the coral alloimmune maturation system may be used as a new evolutionary model scheme for studying tissue transplantation and tolerance.

We have studied the role the canonical Wnt pathway plays in hydroid pattern formation during embryonic development and metamorphosis. Transcripts of Wnt and Tcf were asymmetrically deposited in the oocyte and subsequent developmental stages, marking the sites of first cleavage, posterior larval pole and the upcoming head of the metamorphosed polyp. To address the function of these genes, we activated downstream events of the Wnt pathway by pharmacologically blocking GSK-3beta. These treatments rendered the polar expression of Tcf ubiquitous and induced development of ectopic axes that contained head structures. These results allow concluding that Wnt signaling controls axis formation and regional tissue fates along it, determining one single axis terminus from which later the mouth and hypostome develop. Our data also indicate Wnt functions in axis formation and axial patterning as in higher metazoans, and thus point to an ancestral role of Wnt signaling in these processes in animal evolution.