Mechanical adaptability of sea cucumber Cuvierian tubules involves a mutable collagenous tissue.

Despite their soft body and slow motion, sea cucumbers present a low predation rate, reflecting the presence of efficient defence systems. For instance, members of the family Holothuriidae rely on Cuvierian tubules for their defence. These tubules are normally stored in the posterior coelomic cavity of the animal, but when the sea cucumber is threatened by a potential predator, they are expelled through the cloacal aperture, elongate, become sticky and entangle and immobilise the predator in a matter of seconds. The mechanical properties (extensibility, tensile strength, stiffness and toughness) of quiescent (i.e. in the body cavity) and elongated (i.e. after expulsion) Cuvierian tubules were investigated in the species Holothuria forskali using traction tests. Important mechanical differences were measured between the two types of tubules, reflecting adaptability to their operating mode: to ease elongation, quiescent tubules present a low resistance to extension, while elongated tubules present a high toughness to resist tractions generated by the predator. We demonstrate that a mutable collagenous tissue (MCT) is involved in the functioning of these organs: (1) some mechanical properties of Cuvierian tubules are modified by incubation in a cell-disrupting solution; (2) the connective tissue layer encloses juxtaligamental-like cells, a cell type present in all MCTs; and (3) tensilin, a MCT stiffening protein, was localised inside these cells. Cuvierian tubules thus appear to enclose a new type of MCT which shows irreversible stiffening.

Instantaneous adhesion of Cuvierian tubules in the sea cucumber Holothuria forskali.

The peculiar Cuvierian tubules of sea cucumbers function as a defense mechanism. They thwart attacks by creating a sticky network composed of elongated tubules within which the potential predator is entangled in a matter of seconds and thus immobilized. Cuvierian tubules are typical instantaneous adhesive organs in which tissue integrity is destroyed during the release of the adhesive secretion. However, very little information is available about this adhesion process. The adhesive epithelium-the mesothelium-and the sticky material it produces were studied in the species Holothuria forskali using different microscopy techniques (light microscopy, transmission electron microscopy, scanning electron microscopy, and atomic force microscopy). The mesothelium consists of two cell types-peritoneocytes and granular cells-organized in superimposed layers. In tubules before expulsion, peritoneocytes form an outer protective cell layer preventing adhesion when not needed. After expulsion, the elongation process removes this protective layer and allows granular cells to unfold and to become exposed at the tubule surface. At this stage, Cuvierian tubules are still not sticky. Upon contact with a surface, however, granular cells release their granule contents. Once released, this material changes in aspect, swells, and spreads readily on any type of substrate where it forms a thin homogeneous layer. After tubule peeling, this layer remains on the surface but is often contaminated with collagen fibers. Atomic force microscopy demonstrated the adhesive layer to be made up of globular nanostructures measuring about 70 nm in diameter and to be more adhesive than the collagen fibers left on it. The morphological organization of Cuvierian tubules therefore allows contact-dependent deposition of an adhesive material presenting a high affinity for various surfaces. It is certainly an adaptive advantage for a defense organ to be able to entangle different types of predators.

Sea star tenacity mediated by a protein that fragments, then aggregates.

Sea stars adhere firmly but temporarily to various substrata as a result of underwater efficient adhesive secretions released by their tube feet. Previous studies showed that this material is mainly made up of proteins, which play a key role in its adhesiveness and cohesiveness. Recently, we solubilized the majority of these proteins and obtained 43 de novo-generated peptide sequences by tandem MS. Here, one of these sequences served to recover the full-length sequence of Sea star footprint protein 1 (Sfp1), by RT-PCR and tube foot transcriptome analysis. Sfp1, a large protein of 3,853 aa, is the second most abundant constituent of the secreted adhesive. By using MS and Western blot analyses, we showed that Sfp1 is translated from a single mRNA and then cleaved into four subunits linked together by disulphide bridges in tube foot adhesive cells. The four subunits display specific protein-, carbohydrate-, and metal-binding domains. Immunohistochemistry and immunocytochemistry located Sfp1 in granules stockpiled by one of the two types of adhesive cells responsible for the secretion of the adhesive material. We also demonstrated that Sfp1 makes up the structural scaffold of the adhesive footprint that remains on the substratum after tube foot detachment. Taken together, the results suggest that Sfp1 is a major structural protein involved in footprint cohesion and possibly in adhesive interactions with the tube foot surface. In recombinant form, it could be used for the design of novel sea star-inspired biomaterials.