The competitive dominance of mussels in the wave-swept rocky intertidal zone is in part due to their ability to maintain a secure attachment. Mussels are tethered to the substratum by a byssus composed of numerous extracellular, collagenous threads secreted by the foot. Each byssal thread has three serially arranged parts: a corrugated proximal region, a smooth distal region and an adhesive plaque. This study examines the material and structural properties of the byssal threads of three mussel species: Mytilus californianus, M. trossulus, and M. galloprovincialis. Tensile tests in general reveal similar material properties among species: the proximal region has a lower initial modulus, a lower ultimate stress and a higher ultimate strain than the distal region. The distal region also yields at a stress well below its ultimate value. In whole thread tests, the proximal region and adhesive plaque are common sites of structural failure and are closely matched in strength, while the distal region appears to be excessively strong. We propose that the high strength of the distal region is the byproduct of a material designed to yield and extend before structural failure occurs. Experimental and theoretical evidence is presented suggesting that thread yield and extensibility provide two important mechanisms for increasing the overall attachment strength of the mussel: (1) the reorientation of threads towards the direction of applied load, and (2) the 'recruitment' of more threads into tension and the consequent distribution of applied load over a larger cross-sectional area, thereby reducing the stress on each thread. This distal region yield behavior is most striking for M. californianus and may be a key to its success in extreme wave-swept environments.

/pdf/mechanical-design-of-mussel-byssus-material-yield-enhances-3fj52r1aof.pdf

Mechanical design of mussel byssus: material yield enhances attachment strength

The peculiar collagens of mussel byssus

Most organisms consist of a functionally adaptive assemblage of hard and soft tissues. Despite the obvious advantages of reinforcing soft protoplasm with a hard scaffold, such composites can lead to tremendous mechanical stresses where the two meet. Although little is known about how nature relieves these stresses, it is generally agreed that fundamental insights about molecular adaptation at hard/soft interfaces could profoundly influence how we think about biomaterials. Based on two noncellular tissues, mussel byssus and polychaete jaws, recent studies suggest that one natural strategy to minimize interfacial stresses between adjoining stiff and soft tissue appears to be the creation of a "fuzzy" boundary, which avoids abrupt changes in mechanical properties. Instead there is a gradual mechanical change that accompanies the transcendence from stiff to soft and vice versa. In byssal threads, the biochemical medium for achieving such a gradual mechanical change involves the elegant use of collagen-based self-assembling block copolymers. There are three distinct diblock copolymer types in which one block is always collagenous, whereas the other can be either elastin-like (soft), amorphous polyglycine (intermediate), or silk-like (stiff). Gradients of these are made by an incrementally titrated expression of the three proteins in secretory cells the titration phenotype of which is linked to their location. Thus, reflecting exactly the composition of each thread, the distal cells secrete primarily the silk- and polyglycine-collagen diblocks, whereas the proximal cells secrete the elastin- and polyglycine-collagen diblocks. Those cells in between exhibit gradations of collagens with silk or elastin blocks. Spontaneous self-assembly appears to be by pH triggered metal binding by histidine (HIS)-rich sequences at both the amino and carboxy termini of the diblocks. In the polychaete jaws, HIS-rich sequences are expanded into a major block domain. Histidine predominates at over 20 mol % near the distal tip and diminishes to about 5 mol % near the proximal base. The abundance of histidine is directly correlated to transition metal content (Zn or Cu) as well as hardness determined by nanoindentation. EXAFS analyses of the jaws indicate that transition metals such as Zn are directly bound to histidine ligands and may serve as cross-linkers.

/pdf/exploring-molecular-and-mechanical-gradients-in-structural-1btr6ikq2q.pdf

Exploring Molecular and Mechanical Gradients in Structural Bioscaffolds

Yield and post-yield behavior of mussel byssal thread: a self-healing biomolecular material.

Most organisms consist of a functionally adaptive assemblage of hard and soft tissues. Despite the obvious advantages of reinforcing soft protoplasm with a hard scaffold, such composites can lead to tremendous mechanical stresses where the two meet. Although little is known about how nature relieves these stresses, it is generally agreed that fundamental insights about molecular adaptation at hard/soft interfaces could profoundly influence how we think about biomaterials. Based on two noncellular tissues, mussel byssus and polychaete jaws, recent studies suggest that one natural strategy to minimize interfacial stresses between adjoining stiff and soft tissue appears to be the creation of a "fuzzy" boundary, which avoids abrupt changes in mechanical properties. Instead there is a gradual mechanical change that accompanies the transcendence from stiff to soft and vice versa. In byssal threads, the biochemical medium for achieving such a gradual mechanical change involves the elegant use of collagen-based self-assembling block copolymers. There are three distinct diblock copolymer types in which one block is always collagenous, whereas the other can be either elastin-like (soft), amorphous polyglycine (intermediate), or silk-like (stiff). Gradients of these are made by an incrementally titrated expression of the three proteins in secretory cells the titration phenotype of which is linked to their location. Thus, reflecting exactly the composition of each thread, the distal cells secrete primarily the silk - and polyglycine-collagen diblocks, whereas the proximal cells secrete the elastin- and polyglycine-collagen diblocks. Those cells in between exhibit gradations of collagens with silk or elastin blocks. Spontaneous self-assembly appears to be by pH triggered metal binding by histidine (HIS)-rich sequences at both the amino and carboxy termini of the diblocks. In the polychaete jaws, HIS-rich sequences are expanded into a major block domain. Histidine predominates at over 20 mol % near the distal tip and diminishes to about 5 mol % near the proximal base. The abundance of histidine is directly correlated to transition metal content (Zn or Cu) as well as hardness determined by nanoindentation. EXAFS analyses of the jaws indicate that transition metals such as Zn are directly bound to histidine ligands and may serve as cross-linkers.

/pdf/current-topics-exploring-molecular-and-mechanical-gradients-geh3g6ntxg.pdf

Current Topics Exploring Molecular and Mechanical Gradients in Structural Bioscaffolds

The ultrastructure of the byssus of Mytilus galloprovincialis was analysed by transmission electron microscopy in thin sections of either embedded or frozen samples. All parts of the byssus (stem core laminae, stem outer laminae, threads proximal and distal parts) appear to be formed by the same basic filamentous components organized in different ways at the submicroscopic level and embedded in a variable quantity of matrix. The filaments appear to consist of a central electron-lucent zone (3 nm in diameter), surrounded by an electron-dense rim (total diameter 7 nm). The matrix has a granular or microfilamentous structure. The stem and the threads differ greatly in their submicroscopic organization, but their basic constituents (filaments and matrix) are similar. Peculiar filamentous banded elements (FBE) were found mainly in the stem outer laminae. A relation between the ultrastructure and mechanical properties of the different parts of the byssus was established. The presence of collagen is discussed; since no morphological evidence of any of the known forms of collagen organization was revealed by electron microscopy, it is suggested that byssus collagen may be localized in the matrix and in the FBE.

The ultrastructure of the byssal apparatus of Mytilus galloprovincialis. IV. Observations by transmission electron microscopy.

Calorimetric, biochemical and morphological investigations to validate a restoration method of fire injured ancient parchment

Sections of integument from gastropod, bivalve and cephalopod species were studied immunohistochemically to determine reactivity to antibody against the type I-like collagen from Sepia cartilage and antibodies against components of the extracellular matrix (ECM) of vertebrate connective tissue: type I, III, IV, V, and VI collagens, laminin, nidogen and heparan sulphate. All samples exhibited similar reactivities to the antibodies, although differences in the intensity and localization of the immunostaining were found that were clearly correlated with between-species differences in integumental ultrastructure. These findings indicate that the composition of the integumental ECM is similar in the three classes of molluscs examined and that several types of collagen are present. However molluscan subepidermal connective tissue differs from the ECM of vertebrate dermis: molluscan integumental ECM contains collagens similar to type I, V and VI collagens but has no type III-similar collagen. Furthermore molecules similar to the type IV collagen, laminin, nidogen and heparan sulphate of vertebrates were present ubiquitously in molluscan basement membrane, confirming the statement that the structure and composition of basement membrane have remained constant throughout the evolution of all animal phyla.

/pdf/immunohistochemical-study-of-subepidermal-connective-of-42cww2e4ly.pdf

Immunohistochemical study of subepidermal connective of molluscan integument.

A substance immunologically related to vertebrate glutamic acid decarboxylase (GAD) has been visualized in the pedal ganglion of Mytilus with the pre-embedding peroxidase-antiperoxidase method, by use of an antiserum raised in sheep against rat brain GAD. The results show that GAD-like immunoreactivity is present both in neuronal perikarya and in nerve fibers. Positive neurons are located mainly among the fibers of the ganglion neuropil at the commissural level, and more rarely close to unreactive cortical cell bodies. Immunoreactive nerve fibers are observed throughout the neuropil and also in cerebropedal and pedal nerves.

L. Vitellaro Zuccarello

Papers

The ultrastructure of the byssal apparatus of Mytilus galloprovincialis. IV. Observations by transmission electron microscopy.

Calorimetric, biochemical and morphological investigations to validate a restoration method of fire injured ancient parchment

Immunohistochemical study of subepidermal connective of molluscan integument.

Glutamic acid decarboxylase (GAD)-like immunoreactivity in the pedal ganglion of Mytilus galloprovincialis