Abstract

BackgroundThe Notch signalling pathway is conserved in pre-bilaterian animals. In the Cnidarian Hydra it is involved in interstitial stem cell differentiation and in boundary formation during budding. Experimental evidence suggests that in Hydra Notch is activated by presenilin through proteolytic cleavage at the S3 site as in all animals. However, the endogenous ligand for HvNotch has not been described yet.ResultsWe have cloned a cDNA from Hydra, which encodes a bona-fide Notch ligand with a conserved domain structure similar to that of Jagged-like Notch ligands from other animals. Hyjagged mRNA is undetectable in adult Hydra by in situ hybridisation but is strongly upregulated and easily visible at the border between bud and parent shortly before bud detachment. In contrast, HyJagged protein is found in all cell types of an adult hydra, where it localises to membranes and endosomes. Co-localisation experiments showed that it is present in the same cells as HvNotch, however not always in the same membrane structures.ConclusionsThe putative Notch ligand HyJagged is conserved in Cnidarians. Together with HvNotch it may be involved in the formation of the parent-bud boundary in Hydra. Moreover, protein distribution of both, HvNotch receptor and HyJagged indicate a more widespread function for these two transmembrane proteins in the adult hydra, which may be regulated by additional factors, possibly involving endocytic pathways.

Highlights

  • The Notch signalling pathway is conserved in pre-bilaterian animals

  • It contains a signal peptide (1-22aa), a DSL (Delta, Serrate, Lag) domain (181-252aa), which is characteristic for Notch ligands (Figure 2) [13], and five EGF repeats (255-540aa)

  • The presence of the von Willebrand type C domain, which is characteristic for Serrate/Jagged-like ligands, led us to name the Hydra protein HyJagged

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Summary

Introduction

The Notch signalling pathway is conserved in pre-bilaterian animals. In the Cnidarian Hydra it is involved in interstitial stem cell differentiation and in boundary formation during budding. Embryonic development is regulated in all animals by common signalling pathways such as the Wnt-, FGF, Hedgehog-, TGFb/BMP- and Notch pathways. They have been shown to be present in basal metazoans such as Cnidarians [1,2,3,4,5]. The fresh water polyp Hydra consists of a simple tube-like body made of two epithelial layers, the inner endoderm and the outer ectoderm separated by an acellular mesoglea It further possesses a hypostome and a ring of tentacles at the apical end and a foot at its basal end. They constitute a pluripotent stem cell lineage that differentiates nerve cells, gland cells, germ cells and

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