M. Daniela Candia Carnevali

University of Milan, Milano, Lombardy, Italy

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Publications (66)121.4 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Among echinoderms, crinoids are well known for their remarkable regenerative potential. Regeneration depends mainly on progenitor cells (undifferentiated or differentiated), which migrate and proliferate in the lesion site. The crucial role of the "progenitor" elements involved in the regenerative processes, in terms of cell recruitment, sources, and fate, is a central problem in view of its topical interest and biological implications. The spectacular regenerative potential of crinoids is used to replace lost internal and external organs. In particular, the process of arm regeneration in the feather star Antedon mediterranea is the regeneration model most extensively explored to date. We have addressed the morphological and functional characterization of the cell phenotypes responsible for the arm regenerative processes by using an in vitro approach. This represents the first successful attempt to culture cells involved in crinoid regeneration. A comparison of these results with others from previous in vivo investigations confirms the diverse cell types contributing to regeneration and underscores their involvement in migration, proliferation, and dedifferentiation processes.
    Cell and Tissue Research 06/2014; · 3.68 Impact Factor
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    ABSTRACT: Although sponges are still often considered to be simple, inactive animals, both larvae and adults of different species show clear coordination phenomena triggered by extrinsic and intrinsic stimuli. Chondrosia reniformis, a common Mediterranean demosponge, lacks both endogenous siliceous spicules and reinforcing spongin fibers and has a very conspicuous collagenous mesohyl. Although this species can stiffen its body in response to mechanical stimulation when handled, almost no quantitative data are available in the literature on this phenomenon. The present work was intended to quantify the dynamic response to mechanical stimulation both of intact animals and isolated tissue samples in order to evaluate: (i) the magnitude of stiffening; (ii) the relationship between the amount of stimulation and the magnitude of the stiffening response; (iii) the ability of the whole body to react to localized stimulation; (iv) the possible occurrence of a conduction mechanism and the role of the exopinacoderm (outer epithelium). Data on mesohyl tensility obtained with mechanical tests confirmed the difference between stimulated and non-stimulated isolated tissue samples, showing a significant relationship between ectosome stiffness and the amount of mechanical stimulation. Our experiments revealed a significant difference in tensility between undisturbed and maximally stiffened sponges and evidence of signal transmission that requires a continuous exopinacoderm. We also provide further evidence for the presence of a chemical factor that alters the interaction between collagen fibrils, thereby changing the mechanical properties of the mesohyl.
    Zoology 01/2014; · 1.47 Impact Factor
  • Silvia Mercurio, Cristiano Di Benedetto, Michela Sugni, M Daniela Candia Carnevali
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    ABSTRACT: In the present work, primary cell cultures from ovaries of the edible sea urchin Paracentrotus lividus were developed in order to provide a simple and versatile experimental tool for researches in echinoderm reproductive biology. Ovary cell phenotypes were identified and characterized by different microscopic techniques. Although cell cultures could be produced from ovaries at all stages of maturation, the cells appeared healthier and viable, displaying a higher survival rate, when ovaries at early stages of gametogenesis were used. In terms of culture medium, ovarian cells were successfully cultured in modified Leibovitz-15 medium, whereas poor results were obtained in minimum essential medium Eagle and medium 199. Different substrates were tested, but ovarian cells completely adhered only on poly-L-lysine. To improve in vitro conditions and stimulate cell proliferation, different serum-supplements were tested. Fetal calf serum and an originally developed pluteus extract were detrimental to cell survival, apparently accelerating processes of cell death. In contrast, cells cultured with sea urchin egg extract appeared larger and healthier, displaying an increased longevity that allowed maintaining them for up to 1 month. Overall, our study provides new experimental bases and procedures for producing successfully long-term primary cell cultures from sea urchin ovaries offering a good potential to study echinoid oogenesis in a controlled system and to investigate different aspects of echinoderm endocrinology and reproductive biology.
    In Vitro Cellular & Developmental Biology - Animal 09/2013; · 1.29 Impact Factor
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    ABSTRACT: Echinoderms and sponges share a unique feature that helps them face predators and other environmental pressures. They both possess collagenous tissues with adaptable viscoelastic properties. In terms of morphology these structures are typical connective tissues containing collagen fibrils, fibroblast- and fibroclast-like cells, as well as unusual components such as, in echinoderms, neurosecretory-like cells that receive motor innervation. The mechanisms underpinning the adaptability of these tissues are not completely understood. Biomechanical changes can lead to an abrupt increase in stiffness (increasing protection against predation) or to the detachment of body parts (in response to a predator or to adverse environmental conditions) that are regenerated. Apart from these advantages, the responsiveness of echinoderm and sponge collagenous tissues to ionic composition and temperature makes them potentially vulnerable to global environmental changes.
    Marine environmental research 08/2013; · 2.34 Impact Factor
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    ABSTRACT: 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.
    Biointerphases 12/2012; 7(1-4):38. · 1.91 Impact Factor
  • Iain C Wilkie, Alice Barbaglio, M Daniela Candia Carnevali
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    ABSTRACT: Although l-glutamate is the most widespread excitatory neurotransmitter in vertebrate and invertebrate nervous systems, there is only sparse evidence that it has this role in echinoderms. Following our previous finding that l-glutamate is widely distributed in the arms of the featherstar (crinoid echinoderm) Antedon mediterranea and initiates arm autotomy (defensive detachment), we now provide evidence of glutamatergic involvement in the control of the arm muscles of the same species using immunocytochemical and physiological methods. Immunofluorescence and immunoenzymatic techniques, which employed the same polyclonal antibody against l-glutamate conjugated to glutaraldehyde, revealed a high level of glutamate-like reactivity in the brachial muscles. By recording the mechanical responses of isolated arm pieces, we found that l-glutamate, l-aspartate and elevated [K(+)](o) induced rhythmic muscle contractions, while glycine, γ-aminobutyric acid, adrenaline and acetylcholine had either no, or no consistent, effect. The frequency and duration of the dominant component of the rhythmic contractions indicated that these may be responsible for the rhythmic activity of the arms that occurs during swimming and after autotomy. We conclude that it is highly likely that l-glutamate has at least a neuromodulatory role in the neural pathways controlling the brachial muscles of A. mediterranea.
    Zoology 11/2012; · 1.47 Impact Factor
  • Michela Sugni, Daphne Motta, Paolo Tremolada, Maria Daniela Candia Carnevali
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    ABSTRACT: Although several authors have suggested a plausible involvement of steroids in the reproductive biology of echinoderms, their definitive role is still poorly understood. In this paper we focused on oestradiol (E2), whose presence and variations were previously revealed in different echinoderm tissues. The aim of this investigation was to provide further information on the scarcely known role of this hormone in the reproductive biology of sea urchins. We injected two different concentrations (5 ng ml−1 and 50 ng ml−1) of 17ß-oestradiol into specimens of the common Paracentrotus lividus for 10 weeks. The E2 treatment did not influence the maturation stage of the gonads and the development of the gametes; it caused a slight decrease in the gonad index and an increase in lipid content. Our present results suggest that E2 could have a function different from that reported for vertebrates and suggested for other echinoderms such as asteroids.
    Journal of the Marine Biological Association of the UK 09/2012; 92(06). · 1.02 Impact Factor
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    ABSTRACT: Echinoderms have been used often as experimental models in developmental biology and evolutionary biology studies. Numerous data on echinoid and asteroid development are available, whereas little is known about crinoid larval biology. This contribution focussed on the life cycle of the Mediterranean feather star Antedon mediterranea. Light and electron microscopy were used to characterize, in detail, the morphology and behaviour of the main larval stages. Similarities and differences with respect to what is already known for other crinoids, and echinoderm species, were explored. In view of the importance of serotonin during settlement and morphogenesis, analyses of the distribution of this molecule were carried out on swimming larvae. Immunolabelling results suggested a role for serotonin in A. mediterranea development, underlining the ancestral importance of this conserved neurotransmitter in deuterostome evolution.
    Invertebrate Reproduction and Development 06/2012; 56(2):124-137. · 0.52 Impact Factor
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    ABSTRACT: Mutable collagenous tissues (MCTs) of echinoderms show reversible changes in tensile properties (mutability) that are initiated and modulated by the nervous system via the activities of cells known as juxtaligamental cells. The molecular mechanism underpinning this mechanical adaptability has still to be elucidated. Adaptable connective tissues are also present in mammals, most notably in the uterine cervix, in which changes in stiffness result partly from changes in the balance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). There have been no attempts to assess the potential involvement of MMPs in the echinoderm mutability phenomenon, apart from studies dealing with a process whose relationship to the latter is uncertain. In this investigation we used the compass depressor ligaments (CDLs) of the sea-urchin Paracentrotus lividus. The effect of a synthetic MMP inhibitor - galardin - on the biomechanical properties of CDLs in different mechanical states ("standard", "compliant" and "stiff") was evaluated by dynamic mechanical analysis, and the presence of MMPs in normal and galardin-treated CDLs was determined semi-quantitatively by gelatin zymography. Galardin reversibly increased the stiffness and storage modulus of CDLs in all three states, although its effect was significantly lower in stiff than in standard or compliant CDLs. Gelatin zymography revealed a progressive increase in total gelatinolytic activity between the compliant, standard and stiff states, which was possibly due primarily to higher molecular weight components resulting from the inhibition and degradation of MMPs. Galardin caused no change in the gelatinolytic activity of stiff CDLs, a pronounced and statistically significant reduction in that of standard CDLs, and a pronounced, but not statistically significant, reduction in that of compliant CDLs. Our results provide evidence that MMPs may contribute to the variable tensility of the CDLs, in the light of which we provide an updated hypothesis for the regulatory mechanism controlling MCT mutability.
    PLoS ONE 01/2012; 7(11):e49016. · 3.53 Impact Factor
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    ABSTRACT: This paper provides insights into the achievements and challenges of implementing education on dual-use in four countries: Austria, Italy, Pakistan and Sweden. It draws attention to the different institutional mechanisms through which dual-use education may be introduced into academic curricula and some of the difficulties encountered in this process. It concludes that there is no 'one size fits all' approach to the implementation of dual-use education. Rather, initiatives must be tailored to suit the teaching traditions, geographical and historical context in which they are being delivered. However, a number of common principles and themes can be derived from all four cases. All these courses bring together a number of different topics that place 'dual-use' in the broader context of biosafety, biosecurity, ethics, law and the environment. The case studies suggest that success in this area depends largely on the leadership and commitment of individuals directly involved in teaching, who are active within the scientific community.
    Medicine Conflict and Survival 01/2012; 28(1):31-44.
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    ABSTRACT: Echinoderms possess unique connective tissues, called mutable collagenous tissues (MCTs), which undergo nervously mediated, drastic and reversible or irreversible changes in their mechanical properties. Connective tissue mutability influences all aspects of echinoderm biology and is a key-factor in the ecological success of the phylum. Due to their sensitivity to endogenous or exogenous agents, MCTs may be targets for a number of common pollutants, with potentially drastic effects on vital functions. Besides its ecological relevance, MCT represents a topic with relevance to several applied fields. A promising research route looks at MCTs as a source of inspiration for the development of novel biomaterials. This contribution presents a review of MCT biology, which incorporates recent ultrastructural, biomolecular and biochemical analyses carried out in a biotechnological context.
    Marine environmental research 08/2011; 76:108-13. · 2.34 Impact Factor
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    ABSTRACT: The mutable collagenous tissue (MCT) of echinoderms has the ability to undergo rapid and reversible changes in passive mechanical properties that are initiated and modulated by the nervous system. Since the mechanism of MCT mutability is poorly understood, the aim of this work was to provide a detailed morphological analysis of a typical mutable collagenous structure in its different mechanical states. The model studied was the compass depressor ligament (CDL) of a sea urchin (Paracentrotus lividus), which was characterized in different functional states mimicking MCT mutability. Transmission electron microscopy, histochemistry, cryo-scanning electron microscopy, focused ion beam/scanning electron microscopy, and field emission gun-environmental scanning electron microscopy were used to visualize CDLs at the micro- and nano-scales. This investigation has revealed previously unreported differences in both extracellular and cellular constituents, expanding the current knowledge of the relationship between the organization of the CDL and its mechanical state. Scanning electron microscopies in particular provided a three-dimensional overview of CDL architecture at the micro- and nano-scales, and clarified the micro-organization of the ECM components that are involved in mutability. Further evidence that the juxtaligamental cells are the effectors of these changes in mechanical properties was provided by a correlation between their cytology and the tensile state of the CDLs.
    PLoS ONE 01/2011; 6(9):e24822. · 3.53 Impact Factor
  • M Daniela Candia Carnevali, Paolo Burighel
    09/2010; , ISBN: 9780470015902
  • I C Wilkie, A Barbaglio, W M Maclaren, M D Candia Carnevali
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    ABSTRACT: 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.
    Journal of Experimental Biology 06/2010; 213(Pt 12):2104-15. · 3.24 Impact Factor
  • Michela Sugni, Iain C. Wilkie, Paolo Burighel, M. Daniela Candia Carnevali
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    ABSTRACT: As well as acting as a neurotransmitter, serotonin is also involved in morphogenetic signalling during crucial phases of many developmental and regenerative processes such as cleavage, migration and differentiation. Echinoderms display nerve-dependent regenerative phenomena and serotonin is one of the main neural regulatory factors that have been identified in them. The present work was designed to investigate the broad-spectrum involvement of this molecule in echinoderm regeneration, focusing on arm regeneration in the crinoid Antedon mediterranea. We carried out specific in vivo exposure experiments to selected anti-serotonergic drugs (a synthesis inhibitor: parachlorophenylalanine and a receptor antagonist: methiothepin) and explored their possible effects on arm regeneration. Drug exposure appeared to affect regeneration, causing a general delay of regenerative growth and several histological anomalies mainly in the muscles of the stump. In addition, exposure to the antagonist produced a marked reduction of cell proliferation in both the regenerate and the stump. Our results provide further evidence that serotonin has wider functions than those related to interneuronal communication and that it may be a critical signalling molecule in the main processes of regeneration such as proliferation, morphogenesis and differentiation.
    Journal of the Marine Biological Association of the UK 04/2010; 90(03):555 - 562. · 1.02 Impact Factor
  • Francesco Bonasoro, M. Daniela Candia Carnevali
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    ABSTRACT: Regular sea–urchins, as a rule, prepare and process their food as pellets in their buccal cavity before ingesting them. This is obtained through the synergistic interaction of the five teeth with five fleshy protuberances (paradental tongues) which extend into the oral cavity and work like tongues. Each of these structures consists of two anatomically distinct parts: the outer component is a deep pouch of the terminal tract of the pharynx and shows a histological organization similar to that usually shown by the pharyngeal wall itself. The deeper component (paradental axis) is represented by a supporting rod and shows a peculiar histological structure, which consists of glycogen containing vesiculate cells mixed to slim muscle fibres and held together by a thin fibrillar stroma. The paradental axes are very problematic structures in terms of comparative anatomy and phylogenetic aspects. Their microscopic and submicroscopic organization is unusual for an echinoderm and recalls that of different types of chordoid organs commonly found in other invertebrates (Protostomata and Deuterostomata).
    Acta Zoologica 04/2010; 75(2):89 - 100. · 1.47 Impact Factor
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    ABSTRACT: Two echinoderm species, the sea urchin Paracentrotus lividus and the feather star Antedon mediterranea, were exposed for 28 days to several EDCs: three putative androgenic compounds, triphenyltin (TPT), fenarimol (FEN), methyltestosterone (MET), and two putative antiandrogenic compounds, p,p'-DDE (DDE) and cyproterone acetate (CPA). The exposure nominal concentrations were from 10 to 3000 ng L(-1), depending on the compound. This paper is an attempt to join three different aspects coming from our ecotoxicological tests: (1) the chemical behaviour inside the experimental system; (2) the measured toxicological endpoints; (3) the biochemical responses, to which the measured endpoints may depend. The chemical fate of the different compounds was enquired by a modelling approach throughout the application of the 'Aquarium model'. An estimation of the day-to-day concentration levels in water and biota were obtained together with the amount assumed each day by each animal (uptake in microg animal(-1) d(-1) or ng g-wet weight(-1) d(-1)). The toxicological endpoints investigated deal with the reproductive potential (gonad maturation stage, gonad index and oocyte diameter) and with the regenerative potential (growth and histology). Almost all the compounds exerted some kind of effect at the tested concentrations, however TPT was the most effective in altering both reproductive and regenerative parameters (also at the concentration of few ng L(-1)). The biochemical analyses of testosterone (T) and 17beta-estradiol (E(2)) also showed the ability of the selected compounds to significantly alter endogenous steroid concentrations.
    Ecotoxicology 11/2009; 19(3):538-54. · 2.77 Impact Factor
  • I. C. Wilkie, M. D. Candia Carnevali, F. Andrietti
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    ABSTRACT: The peristomial membrane (PM) is the area of flexible body wall which surrounds the mouth of regular sea-urchins. In cidaroid sea-urchins, like Stylocidaris. affnis (Phil.), it contains an extensive endoskeleton of overlapping plates. This paper describes the basic mechanical properties of the PM of S. affinis and compares them with those of the PM of the previously studied echinacean euechinoid Paracentrotus lividus. When subjected to vertical deformation, the PM of S. affinis generates J-shaped tension-deformation curves with an inflexion point at a deformation corresponding to about 3% of the perignathic girdle diameter. It is considerably stiffer than the P. lividus PM and, unlike the latter, it shows asymmetrical properties, i.e. greater resistance to aboral flexion (retraction) than to oral flexion (protraction). In interambulacral regions the junction between the PM endoskeleton and test of S. affinis takes the form of a wide ligament-filled gap which acts as a compliant hinge. In marked contrast to the echinacean PM, that of S. affinis provides little evidence for the presence of mutable collagenous tissue (MCT). The cidaroid PM is interpreted as a design which maximizes mechanical defence whilst retaining the capacity for some flexibility. A possible correlation between the lantern systems and feeding habits of sea-urchins is discussed.
    Journal of Zoology 05/2009; 238(3):557 - 569. · 2.04 Impact Factor
  • F. Andrietti, M. D. Candia Carnevali, I. C. Wilkie
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    ABSTRACT: In the present analysis experimental results and computer modelling have been used to compare biomechanics of the two most contrasting lantern types among regular echinoids, i.e. the cidarid-type (Stylocidaris affinis) and the camarodont (Paracentrotus lividus) lantern. The lantern is modelled as a rigid pyramid anchored in the centre of the peristomial area. Each side is connected to the test by means of muscles (protractors and retractors) rind ligaments (compass depressors). The overall movement of the lantern may be resolved into vertical displacement and lateral tilting. Computer-aided mechanical analysis of the system takes into account the action of protractor and/or retractor muscles, their elastic reaction and the interaction with the ligamentous structures (peristomial membrane and compass depressors). The geometrical parameters have been determined by preliminary structural analysis and the values of active and passive forces have been measured experimentally. The results of simulations highlight important differences in the mechanics of the Stylocidaris lantern compared to that of Paracentrotus: (1) a very much greater antagonistic effect of muscular passive stretch resistance on lateral tilting: (2) a minor role of the peristomial membrane tensile strength and muscular activity in controlling vertical lantern displacement: (3) a less efficient lantern retraction and protraction owing to unfavourable muscle insertion position on the perignathic girdle and ineffective muscular contraction forces. As a conclusion, in Stylocidaris the action of the protractor and retractor muscles seems to play no significant role in moving the overall lantern, their action being mainly exerted to stabilize it. On the contrary, in Paracentrotus such stabilizing action is provided by the compass depressor ligaments. Finally, on theoretical grounds the arrangement of muscle attachments around the perignathic girdle in Stylocidaris seems to be just as good as that of Paracentrotus for controlling overall lantern mobility, if relative heights of insertions are ignored.
    Journal of Zoology 03/2009; 231(4):595 - 610. · 2.04 Impact Factor
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    ABSTRACT: The dental apparatus or Aristotle's lantern of sea-urchins is a complex system of interacting skeletal ossicles, joints, muscles and ligaments arranged in a rigorous geometry and involved in a variety of activities. In this paper we study the movement of the whole lantern system modelled as a rigid body. The model lantern is constrained at its apex by the peristomial membrane and its movement is controlled by five pairs of antagonistic forces (retractor and protractor muscles). The other main forces applied to the lantern are the elastic reactions of both muscles and ligamental structures (compass depressors and peristomial membrane). The lantern is allowed to perform vertical movements and lateral inclinations but cannot rotate around its main axis. The equilibrium conditions of the system have been found by means of a numerical iterative procedure for solving non-linear equations. The results of the present analysis allow simulation of the overall mechanical activity of the lantern taking into account the experimental data regarding active and passive muscular forces and the tensile constraints due to ligaments.
    Journal of Zoology 03/2009; 220(3):345 - 366. · 2.04 Impact Factor

Publication Stats

515 Citations
121.40 Total Impact Points

Institutions

  • 1979–2014
    • University of Milan
      • • Department of Life Sciences
      • • Department of Anesthesia, Intensive Care and Dermatologic Sciences
      Milano, Lombardy, Italy
  • 2006–2012
    • Glasgow Caledonian University
      • Division of Biomedical Sciences
      Glasgow, SCT, United Kingdom
  • 2010
    • University of Padova
      • Department of Biology
      Padua, Veneto, Italy