Correlations between the biochemistry and mechanical states of a sea-urchin ligament: a mutable collagenous structure.

Mutable collagenous tissues (MCTs) of echinoderms can be regarded as intelligent and dynamic biomaterials, due to their ability to reversibly change their mechanical properties in a short physiological time span. This mutability phenomenon is nervously mediated and involves secreted factors of the specialized 'juxtaligamental' cells, which, when released into the extracellular matrix (ECM), change the cohesive forces between collagen fibrils. MCTs exist in nature in several forms, including some associated with echinoderm autotomy mechanisms. Since the molecular mechanism of mutability is still incompletely understood, the aim of this work was to provide a detailed biochemical analysis of a typical mutable collagenous structure and to identify possible correlations between its biochemistry and mechanical states. A better understanding of the mutability phenomena is likely to provide a unique opportunity to develop new concepts that can be applied in the design of dynamic biomaterial for tissue regeneration, leading to new strategies in regenerative medicine. The MCT model used was the compass depressor ligament (CDL) of a sea urchin (Paracentrotus lividus), which was analyzed in different mechanical states, mimicking the mutability phenomenon. Spectroscopic techniques, namely Fourier transform infrared (FT-IR) and confocal Raman microscopy, were used to identify the specific molecular components that contribute to the CDL biochemical microenvironment and to investigate the possibility that remodelling/synthesis of new ECM components occurs during the mutability phenomenon by analogy with events during pregnancy in the uterine cervix of mammals (which also consists mainly of mechanically adaptable connective tissues). The results demonstrate that CDL ECM includes collagen with biochemical similarities to mammalian type I collagen, as well as sulphated glycosaminoglycans (GAGs). CDL mutability seems to involve a molecular rearrangement of the ECM, without synthesis of new ECM components. Although there were no significant biochemical differences between CDLs in the various mechanical states were observed. However, subtle adjustments in tissue hydration seemed to occur, particularly during stiffening.

Physiological and immunocytochemical evidence that glutamatergic neurotransmission is involved in the activation of arm autotomy in the featherstar Antedon mediterranea (Echinodermata: Crinoidea).

The crinoid echinoderm Antedon mediterranea autotomises its arms at specialised skeletal joints known as syzygies that occur at regular intervals along the length of each arm. Detachment is achieved through the nervously mediated destabilisation of ligament fibres at a particular syzygy. The aim of this investigation was to identify neurotransmitters that are involved in the autotomy response. Physiological experiments were conducted on isolated preparations of syzygial joints, which can be induced to undergo autotomy-like fracture by applying stimulatory agents such as elevated [K(+)](o). Initial experiments with elevated [K(+)](o) showed that the autotomy threshold (the minimum amount of stimulation required to provoke autotomy) is lowest in syzygies at the arm base and rises distally. Of a range of neurotransmitter agonists tested, only l-glutamate invoked syzygial destabilisation, as did its analogues l-aspartate, alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and kainate, but not l-(+)-2-amino-4-phosphonobutyrate (l-AP4) or N-methyl-d-aspartate (NMDA). The implication that l-glutamate stimulates syzygial fracture through AMPA/kainate-like receptors was supported by the finding that the action of l-glutamate was inhibited by the AMPA/kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Acetylcholine depressed the response of syzygial preparations to l-glutamate, suggesting a possible mechanism by which the autotomy threshold could be varied constitutively and facultatively. An immunocytochemical method employing a polyclonal antibody against l-glutamate conjugated to glutaraldehyde revealed l-glutamate-like immunoreactivity in all components of the putative neural pathway controlling the autotomy reflex, including the epidermis, brachial nerve, syzygial nerves and cellular elements close to the syzygial ligaments. We conclude that it is highly probable that l-glutamate acts as an excitatory neurotransmitter in the activation of arm autotomy in A. mediterranea.

Mechanical adaptability of a sponge extracellular matrix: Evidence for cellular control of mesohyl stiffness in Chondrosia reniformis Nardo.

The marine sponge Chondrosia reniformis Nardo consists largely of a collagenous tissue, the mesohyl, which confers a cartilaginous consistency on the whole animal. This investigation was prompted by the incidental observation that, despite a paucity of potentially contractile elements in the mesohyl, intact C. reniformis stiffen noticeably when touched. By measuring the deflection under gravity of beam-shaped tissue samples, it was demonstrated that the flexural stiffness of the mesohyl is altered by treatments that influence cellular activities, including [Ca2+] manipulation, inorganic and organic calcium channel-blockers and cell membrane disrupters, and that it is also sensitive to extracts of C. reniformis tissue that have been repeatedly frozen then thawed. Since the membrane disrupters and tissue extracts cause marked stiffening of mesohyl samples, it is hypothesised that cells in the mesohyl store a stiffening factor and that the physiologically controlled release of this factor is responsible for the touch-induced stiffening of intact animals.

Mutable collagenous tissue: overview and biotechnological perspective.

The mutable collagenous tissue (MCT) of echinoderms can undergo extreme changes in passive mechanical properties within a timescale of less than 1 s to a few minutes, involving a mechanism that is under direct neural control and coordinated with the activities of muscles. MCT occurs at a variety of anatomical locations in all echinoderm classes, is involved in every investigated echinoderm autotomy mechanism, and provides a mechanism for the energy-sparing maintenance of posture. It is therefore crucially important for the biology of extant echinoderms. This chapter summarises current knowledge of the physiology and organisation of MCT, with particular attention being given to its molecular organisation and the molecular mechanism of mutability. The biotechnological potential of MCT is discussed. It is argued that MCT could be a source of, or inspiration for, (1) new pharmacological agents and strategies designed to manipulate therapeutically connective tissue mechanical properties and (2) new composite materials with biomedical applications.

Is muscle involved in the mechanical adaptability of echinoderm mutable collagenous tissue?

The mutable collagenous tissue (MCT) of echinoderms has the capacity to change its mechanical properties in a time scale of less than 1 s to a few minutes under the influence of the nervous system. Although accumulating evidence indicates that the mechanical adaptability of MCT is due primarily to the modulation of interactions between components of the extracellular matrix, the presence of muscle in a few mutable collagenous structures has led some workers to suggest that contractile cells may play an important role in the phenomenon of variable tensility and to call for a re-evaluation of the whole MCT concept. This contribution summarises present information on MCT and appraises the argument implicating muscle in its unique mechanical behaviour. It is concluded that there is no evidence that the variability of the passive mechanical properties of any mutable collagenous structure is due to muscle.

Autotomy as a prelude to regeneration in echinoderms.

'Autotomy' refers to the adaptive detachment of animal body parts where this serves a defensive function, is achieved by an intrinsic mechanism, and is nervously mediated. With regard to each echinoderm class, this article itemises those structures that are autotomous, evaluates the extent to which autotomy precedes regeneration in natural populations, reviews current knowledge of the morphology of autotomy planes and mechanisms that effect fracture at autotomy, and comments on autotomy-related issues arising from studies of the cellular events of regeneration. Each autotomy plane can be regarded as an assemblage of breakage zones traversing the individual anatomical components of the autotomous structure. In any one autotomy plane some breakage zones are permanent sites of weakness that are fractured by external forces and some are potential sites of weakness that undergo a loss of tensile strength only at the time of autotomy. The latter occur predominantly in mutable collagenous structures, although there are a few examples of muscles that undergo an endogenous rupturing process. Available evidence indicates that autotomy is by far the commonest proximate cause of structural loss in echinoderms. Most echinoderm regeneration is therefore necessitated by autotomy and proceeds from the retained side of a fractured autotomy plane. Due to a lack of relevant research there is as yet little evidence for or against the presence of specific regeneration-promoting adaptations at autotomy planes, although it is argued that an autotomy plane designed primarily to effect rapid detachment would by itself increase regenerative efficiency.

Mutable Collagenous Structure or Not? A Comment on the Re-interpretation by del Castillo et al. of the Catch Mechanism in the Sea Urchin Spine Ligament.

It is now widely recognized that one of the characteristic features of the phylum Echinodermata is the presence of collagenous tissues that can undergo rapid, nervously mediated changes in mechanical properties (1,2). Whilst examples of such mutable collagenous tissues have been demonstrated in the five major echinoderm classes (3,4), research over the past 30 years has focused on a certain few of them that are easily accessible and amenable to experimental investigation. The capsular ligament ("catch apparatus"--CA) of the echinoid spine-test joint is one of these well-studied collagenous structures. Because the CA can undergo reversible changes in passive stifness, it was believed to be a catch muscle by von Uexkull (5). Subsequently it was shown to consist predominantly of collagenous connective tissue with only a scattering of small muscle fibers. Analysis of its mechanical properties and physiological responses led most workers to the conclusion that the CA is a posture-locking device whose functioning depends almost entirely on the variable tensility of its extracellular connective tissue (6-11). However, de1 Castillo et al. (12) have presented a model in which its catch properties are attributed only to the contractile cells and in which connective tissue mutability plays no role. The aims of this contribution are to evaluate the new model, to show that it cannot explain the behavior of the CA, and to demonstrate that the extracellular matrix must be the primary source of the catch properties of the CA.

Pressure sore prevention for the wheelchair-bound spinal injury patient.

The concept of a wheelchair cushion fitting clinic for the prevention of pressure sores is reviewed in the light of recent estimates of the cost of pressure sores in the U.K. A method for measuring the pressure beneath the ischial tuberosities is discussed and techniques for measuring a patient's habitual exercise frequency and seated posture are described. Results from the records of 600 spinal injury patients including Rancho Los Amigos Hospital are reported and used to demonstrate the importance of low pressure beneath the ischial tuberosities as an indicator of wheelchair cushion suitability.

The juxtaligamental cells of Ophiocomina nigra (Abildgaard) (Echinodermata: Ophiuroidea) and their possible role in mechano-effector function of collagenous tissue.

The intervertebral ligament of the brittlestar Ophiocomina nigra contains numerous cellular processes which belong to perikarya located on the outer surfaces of the ligament. These are described as the juxtaligamental cells and have been studied by light and electron microscopy. The cells are mainly concentrated in four pairs of ganglion-like nodes associated with the intervertebral ligament and in similar nodes adjacent to every other major connective tissue component of the arm. Although their histochemistry and ultrastructure indicate a neurosecretory function, they are anomalous in containing unusually large electron-dense granules probably associated with calcium. The ganglion-like nodes are innervated by hyponeural nerves, though synaptic contacts with the juxtaligamental cells have yet to be demonstrated. The function of the cells is discussed and it is suggested that they may be involved in the rapid loss of tensil strength which the intervertebral ligament sustains during arm autotomy. They may achieve this by controlling the availability of Ca2+ ions to the extracellular compartment of the ligament.