Integrins of the starlet sea anemone Nematostella vectensis.

Integrins are extracellular matrix receptors composed of α and ß subunits. Here we describe two α subunits and four ß subunits from the starlet sea anemone Nematostella vectensis. Phylogenetic analysis suggests that the α subunits are most closely related to RGD- and LDV-dependent α subunits of chordates. The ß subunits cluster with the previously described ß integrins of the hard coral Acropora millepora. The expression of one of the α subunits and three of the ß subunits was confirmed by reverse transcription PCR and in situ hybridization. The α subunit is primarily expressed in cells near muscles, by a subset of gastrodermal cells, and in the gonad. The three ß subunits each have distinctive patterns of expression: one is concentrated in the gonad and mesenteric filament, another is found in a subset of cells in the epidermis of the oral region and in a subset of gastrodermal cells in the mesenteries, and a third is expressed widely. Changes in expression were also studied 48 h after horizontal transection by quantitative reverse transcription PCR and in situ hybridization. One of the ß subunits is expressed 8-fold higher during regeneration, and its expression is observed in cells within both the epidermis and the gastrodermis at the site of regeneration. Our observations confirm that complex patterns of integrin expression were already present in basal metazoans. The integrins expressed in the gonads may play roles in mediating sperm-egg interactions in N. vectensis, while others may play a role in regulating proliferation during regeneration.

A thrombospondin in the anthozoan Nematostella vectensis is associated with the nervous system and upregulated during regeneration.

Thrombospondins are multimeric extracellular matrix glycoproteins that play important roles in development, synaptogenesis and wound healing in mammals. We previously identified four putative thrombospondins in the genome of the starlet sea anemone Nematostella vectensis. This study presents the first analysis of these thrombospondins, with the goals of understanding fundamental roles of thrombospondins in the Eumetazoa. Reverse transcriptase PCR showed that each of the N. vectensis thrombospondins (Nv85341, Nv22035, Nv168100 and Nv30790) is transcribed. Three of the four thrombospondins include an RGD or KGD motif in their thrombospondin type 3 repeats at sites equivalent to mammalian thrombospondins, suggesting ancient roles as RGD integrin ligands. Phylogenetic analysis based on the C-terminal regions demonstrated a high level of sequence diversity between N. vectensis thrombospondins. A full-length cDNA sequence was obtained for Nv168100 (NvTSP168100), which has an unusual domain organization. Immunohistochemistry with an antibody to NvTSP168100 revealed labeling of neuron-like cells in the mesoglea of the retractor muscles and the pharynx. In situ hybridization and quantitative PCR showed that NvTSP168100 is upregulated during regeneration. Immunohistochemistry of the area of regeneration identified strong immunostaining of the glycocalyx, the carbohydrate-rich matrix coating the epidermis, and electron microscopy identified changes in glycocalyx organization during regeneration. Thus, N. vectensis thrombospondins share structural features with thrombospondins from mammals and may have roles in the nervous system and in matrix reorganization during regeneration.