From head to tail: the myoseptal system in basal actinopterygians

Experimental studies indicated that myomeres play several functional roles during swimming. Some of the functions in question are thought to change rostrocaudally, e.g., anterior myomeres are thought to generate forces, whereas posterior myomeres are thought to transmit forces. In order to determine whether these putative functions are reflected in myoseptal morphology we carried out an analysis of the myoseptal system that includes epaxial and hypaxial myosepta of all body regions for the first time. We combined clearing and staining, microdissections, polarized light microscopy, SEM technique, and length measurements of myoseptal parts to reveal the spatial arrangement, collagen fiber architecture, and rostrocaudal gradients of myosepta. We included representatives of the four basal actinopterygian clades to evaluate our findings in an evolutionary and in a functional context. Our comparison revealed a set of actinopterygian groundplan features. This includes a set of specifically arranged myoseptal tendons (epineural, epipleural, lateral, and myorhabdoid tendons) in all epaxial and postanal hypaxial myosepta. Only preanal hypaxial myosepta lack tendons and exclusively consist of mediolateral fibers. Laterally, myosepta generally align with the helically wound fibers of the dermis in order not to limit the body's maximum curvature. Medially, the relationship of myosepta to vertebrae clearly differs from a 1:1 relationship: a myoseptum attaches to the anterior margin of a vertebra, turns caudally, and traverses at least three vertebrae in an almost horizontal orientation in all body regions. By this arrangement, horizontal multiple layers of myosepta are formed along the trunk dorsal and ventral to the horizontal septum. Due to their reinforcement by epineural or epipleural tendons, these multiple layers are hypothesized to resist the radial expansion of underlying muscle fibers and thus contribute to modulation of body stiffness. Rostrocaudally, a dorsoventral symmetry of epaxial and hypaxial myosepta in terms of spatial arrangement and collagen fiber architecture is gradually developed towards the postanal region. Furthermore, the rostrocaudal extension of myosepta measured between anterior and posterior cones gradually increases. This myoseptal region is reinforced by longitudinal fibers of lateral tendons. Furthermore, the percentage of connective tissue in a cross section increases. These morphological data indicate that posterior myosepta are equipped for multisegmental force transmission towards the caudal fin. Anteriormost myosepta have reinforced and elongated dorsal posterior cones. They are adequately designed to transmit epaxial muscular forces to the neurocranium in order to cause its elevation during suction feeding.     

The myoseptal system in Chimaera monstrosa: collagenous fiber architecture and its evolution in the gnathostome stem lineage

Recent studies have revealed the 3D morphology and collagen fiber architecture of myosepta in teleostome fishes. Here we present the first data set on the myoseptal structure of a representative of the chondrichthyan clade. We investigate the series of myosepta in the ratfish Chimaera monstrosa (Holocephali) from the anterior to the posterior body using microdissections of cleared and stained specimens, polarized light microscopy of excised myosepta, and histology. The features of the myoseptal system of Chimaera are compared to data from closely related vertebrate groups and are mapped onto a phylogenetic tree to further clarify the characteristics of the myoseptal series in the gnathostome ancestor. The 3D morphology and collagen fiber architecture of the myoseptal series in C. monstrosa resembles that of Teleostomi (Actinopterygii+Sarcopterygii) with regard to several features. Our comparative analysis reveals that some of them have evolved in the gnathostome stem lineage. (1) A series of epineural and epaxial lateral tendons (LTs) along the whole body, and a series of epipleural and hypaxial LTs in the postanal region evolved in the gnathostome stem lineage. (2) The LTs increase in length towards the posterior body (three-fold in Chimaera). Data on Chimaera and some comparative data on actinopterygian fishes indicate that LTs also increase in thickness towards the posterior body, but further data are necessary to test whether this holds true generally. (3) Another conspicuous apomorphic gnathostome feature is represented by multi-layer structures of myosepta. These are formed along the vertebral column by converging medial regions of successive sloping parts of myosepta. (4) The dorsalmost and ventralmost flanking parts of myosepta bear a set of mediolaterally oriented collagen fibers that are present in all gnathostomes but are lacking in outgroups. Preanal hypaxial myosepta are clearly different from epaxial myosepta and postanal hypaxial myosepta in terms of their collagen fiber architecture. In Chimaera, preanal hypaxial myosepta consist of an array of mediolaterally oriented collagen fibers closely resembling the condition in other gnathostome groups and in petromyzontids. Only one series of tendons, the myorhabdoid tendons of the flanking parts of myosepta, have evolved in the stem lineage of Myopterygii (Gnathostomata+Petromyzontida). Similar to LTs, the tendons of this series also increase in length towards the posterior body. In combination with other studies, the present study provides a framework for the design of morphologically based experiments and modeling to further address the function of myosepta and myoseptal tendons in gnathostomes.     

Myoseptal architecture of sarcopterygian fishes and salamanders with special reference to Ambystoma mexicanum

During axial undulatory swimming in fishes and salamanders muscular forces are transmitted to the vertebral axis and to the tail. One of the major components of force transmission is the myoseptal system. The structure of this system is well known in actinopterygian fishes, but has never been addressed in sarcopterygian fishes or salamanders. In this study we describe the spatial arrangement and collagen fiber architecture of myosepta in Latimeria, two dipnoans, and three salamanders in order to gain insight into function and evolution of the myoseptal system in these groups. Salamander myosepta lack prominent cones, and consist of homogenously distributed collagen fibers of various orientations that never form distinct tendons. Fiber orientations are difficult to homologize with those of fish myosepta. The myosepta of Latimeria and dipnoans (Protopterus and Neoceratodus) illustrate that major changes in architecture occurred in the sarcopterygian clade (loss of horizontal septum), in the rhipidistian (dipnoans + tetrapods) clade (loss of epineural and epipleural tendon), and in tetrapods (loss of lateral tendons and myoseptal folding). When compared to fishes, the myosepta of wholly aquatic salamanders (Ambystoma mexicanum, Amphiuma tridactylum, Necturus maculosus) do not have the lateral tendons we suppose serve to transfer muscular forces posteriorly. We propose that alternative structures (most conspicuously present in Ambystoma) perform this function: posteriorly the relative amount of connective tissue increases considerably, and myosepta are disintegrated to horizontal lamellae of connective tissue. The structures thought to be involved in modulation of body stiffness in fishes during swimming are also absent in salamanders. Our data also have implications for the hypothesis that salamander hypaxial myosepta are designed to increase shortening amplification of the hypaxial muscle fibers. The posterior hypaxial myosepta of all three salamander species possess only mediolaterally directed collagen fibers, which would indeed amplify the shortening of the associated muscle.     

Cruising specialists and accelerators--are different types of fish locomotion driven by differently structured myosepta?

Locomotor specialists, such as accelerators and cruisers, have clearly differing body designs. For physical reasons these designs are mutually exclusive, i.e. cruisers necessarily have poor accelerating capabilities and vice versa. For the first time, we examine whether differences in the anatomy of the musculo-tendinous system of the trunk are present in addition to the differences in external body design. We investigated the myoseptal series of two closely related locomotor specialists, the cruiser Scomber scombrus and the accelerator Channa obscura, by microdissections combined with polarized light microscopy and histology. Our comparison includes 3D-morphology of myosepta, spatial arrangement and length of myoseptal tendons, their relation to red and white muscles, rostrocaudal changes in all these aspects and the musculo-tendinous system of the caudal fin. Regarding all these features, Channa has retained the plesiomorphic condition of its actinopterygian ancestor. In contrast, the derived morphology of Scomber is characterized by (i) lateral (LT) and myorhabdoid tendons (MT) that are lengthened to up to 20% of body length (compared to a maximum of 8.2% in Channa), (ii) posterior myoseptal cones that are subsequently linked by horizontal projections of merged LTs and MTs, (iii) an increased area of red muscle fibers that insert to LTs of myosepta, (iv) the reduction of epineural (ENTs) and epipleural tendons (EPTs) that connect backbone and skin, (v) specific caudal tendons that are identified to be serial homologues of LTs and MTs of more anterior myosepta, (vi) and a partial reduction of intrinsic caudal muscles. These results suggest the following functional adaptations in the cruiser Scomber. Red muscle forces may be transmitted through LTs and posterior cones to the prominent tendons of the caudal fin. The length of LTs and the intersegmental connections along the posterior cones may facilitate posterior force transmission and may be correlated with the long propulsive wavelength generally observed in cruising carangiform swimmers. Epineural and epipleural tendons are interpreted to minimize lateral backbone displacement during high body curvatures. This is consistent with the lack of these tendons in Scomber, because high body curvatures are not displayed in stiffer-bodied carangiform swimmers. It remains to be tested whether the specializations revealed in this initial study for Scomber represent general specializations of carangiform swimmers. Taking into account the geometry of myoseptal tendons and the horizontal septum we evaluate how local bending according to beam-theory can be generated by white or red muscle activity in Channa and Scomber. In both species, the musculo-tendinous anatomy of the caudal fin explains the functional asymmetry of the caudal fin that was experimentally revealed in previous studies.     

Evolution of high-performance swimming in sharks: transformations of the musculotendinous system from subcarangiform to thunniform swimmers

In contrast to all other sharks, lamnid sharks perform a specialized fast and continuous "thunniform" type of locomotion, more similar to that of tunas than to any other known shark or bony fish. Within sharks, it has evolved from a subcarangiform mode. Experimental data show that the two swimming modes in sharks differ remarkably in kinematic patterns as well as in muscle activation patterns, but the morphology of the underlying musculotendinous system (red muscles and myosepta) that drives continuous locomotion remains largely unknown. The goal of this study was to identify differences in the musculotendinous system of the two swimming types and to evaluate these differences in an evolutionary context. Three subcarangiform sharks (the velvet belly lantern shark, Etmopterus spinax, the smallspotted catshark, Scyliorhinus canicula, and the blackmouth catshark, Galeus melanostomus) from the two major clades (two galeans, one squalean) and one lamnid shark, the shortfin mako, Isurus oxyrhinchus, were compared with respect to 1) the 3D shape of myomeres and myosepta of different body positions; 2) the tendinous architecture (collagenous fiber pathways) of myosepta from different body positions; and 3) the association of red muscles with myoseptal tendons. Results show that the three subcarangiform sharks are morphologically similar but differ remarkably from the lamnid condition. Moreover, the "subcarangiform" morphology is similar to the condition known from teleostomes. Thus, major features of the "subcarangiform" condition in sharks have evolved early in gnathostome history: Myosepta have one main anterior-pointing cone and two posterior-pointing cones that project into the musculature. Within a single myoseptum cones are connected by longitudinally oriented tendons (the hypaxial and epaxial lateral and myorhabdoid tendons). Mediolaterally oriented tendons (epineural and epipleural tendons; mediolateral fibers) connect vertebral axis and skin. An individual lateral tendon spans only a short distance along the body (a fraction between 0.05 and 0.075 of total length, L, of the shark). This span is similar in all tendons along the body. Red muscles insert into the midregion of the lateral tendons. The shortfin mako differs substantially from this condition in several respects: Red muscles are internalized and separated from white muscles by a sheath of lubricative connective tissue. They insert into the anterior part of the hypaxial lateral tendon. Rostrocaudally, this tendon becomes very distinct and its span increases threefold (0.06L anteriorly to 0.19L posteriorly). Mediolateral fibers do not form distinct epineural/epipleural tendons in the mako. Since our morphological findings are in good accordance with experimental data it seems likely that the thunniform swimming mode has evolved along with the described morphological specializations.     

The musculotendinous system of an anguilliform swimmer: Muscles, myosepta, dermis, and their interconnections in Anguilla rostrata

Eel locomotion is considered typical of the anguilliform swimming mode of elongate fishes and has received substantial attention from various perspectives such as swimming kinematics, hydrodynamics, muscle physiology, and computational modeling. In contrast to the extensive knowledge of swimming mechanics, there is limited knowledge of the internal body morphology, including the body components that contribute to this function. In this study, we conduct a morphological analysis of the collagenous connective tissue system, i.e., the myosepta and skin, and of the red muscle fibers that sustain steady swimming, focusing on the interconnections between these systems, such as the muscle-tendon and myosepta-skin connections. Our aim is twofold: (1) to identify the morphological features that distinguish this anguilliform swimmer from subcarangiform and carangiform swimmers, and (2) to reveal possible pathways of muscular force transmission by the connective tissue in eels. To detect gradual morphological changes along the trunk we investigated anterior (0.4L), midbody (0.6L), and posterior body positions (0.75L) using microdissections, histology, and three-dimensional reconstructions. We find that eel myosepta have a mediolaterally oriented tendon in each the epaxial and hypaxial regions (epineural or epipleural tendon) and two longitudinally oriented tendons (myorhabdoid and lateral). The latter two are relatively short (4.5-5% of body length) and remain uniform along a rostrocaudal gradient. The skin and its connections were additionally analyzed using scanning electron microscopy (SEM). The stratum compactum of the dermis consists of approximately 30 layers of highly ordered collagen fibers of alternating caudodorsal and caudoventral direction, with fiber angles of 60.51 +/- 7.05 degrees (n = 30) and 57.58 +/- 6.92 degrees (n = 30), respectively. Myosepta insert into the collagenous dermis via fiber bundles that pass through the loose connective tissue of the stratum spongiosum of the dermis and either weave into the layers of the stratum compactum (weaving fiber bundles) or traverse the stratum compactum (transverse fiber bundles). These fiber bundles are evenly distributed along the insertion line of the myoseptum. Red muscles insert into lateral and myorhabdoid myoseptal tendons but not into the horizontal septum or dermis. Thus, red muscle forces might be distributed along these tendons but will only be delivered indirectly into the dermis and horizontal septum. The myosepta-dermis connections, however, appear to be too slack for efficient force transmission and collagenous connections between the myosepta and the horizontal septum are at obtuse angles, a morphology that appears inadequate for efficient force transmission. Though the main modes of undulatory locomotion (anguilliform, subcarangiform, and carangiform) have recently been shown to be very similar with respect to their midline kinematics, we are able to distinguish two morphological classes with respect to the shape and tendon architecture of myosepta. Eels are similar to subcarangiform swimmers (e.g., trout) but are substantially different from carangiform swimmers (e.g., mackerel). This information, in addition to data from kinematic and hydrodynamic studies of swimming, shows that features other than midline kinematics (e.g., wake patterns, muscle activation patterns, and morphology) might be better for describing the different swimming modes of fishes.     

The locomotory system of pearlfish Carapus acus: What morphological features are characteristic for highly flexible fishes?

The body curvature displayed by fishes differs remarkably between species. Some nonmuscular features (e.g., number of vertebrae) are known to influence axial flexibility, but we have poor knowledge of the influence of the musculotendinous system (myosepta and muscles). Whereas this system has been described in stiff‐bodied fishes, we have little data on flexible fishes. In this study, we present new data on the musculotendinous system of a highly flexible fish and compare them to existing data on rigid fishes. We use microdissections with polarized light microscopy to study the three‐dimensional anatomy of myoseptal tendons, histology and immunohistology to study the insertion of muscle fiber types into tendons, and μ‐CT scans to study skeletal anatomy. Results are compared with published data from stiff‐bodied fishes. We identify four important morphological differences between stiff‐bodied fishes and Carapus acus: (1) Carapus bears short tendons in the horizontal septum, whereas rigid fishes have elongated tendons. (2) Carapus bears short lateral tendons in its myosepta, whereas stiff‐bodied fishes bear elongated tendons. Because of its short myoseptal tendons, Carapus retains high axial flexibility. In contrast, elongated tendons restrict axial flexibility in rigid fishes but are able to transmit anteriorly generated muscle forces through long tendons down to the tail. (3) Carapus bears distinct epineural and epipleural tendons in its myosepta, whereas these tendons are weak or absent in rigid fishes. As these tendons firmly connect vertebral axis and skin in Carapus, we consider them to constrain lateral displacement of the vertebral axis during extreme body flexures. (4) Ossifications of myoseptal tendons are only present in C. acus and other more flexible fishes but are absent in rigid fishes. The functional reasons for this remain unexplained. J. Morphol., 2012. © 2011 Wiley Periodicals, Inc.     

The musculoskeletal system of the caudal fin in basal Actinopterygii: heterocercy,diphycercy, homocercy

The caudal fin represents the posteriormost region of the vertebrate axis and is one location where forces are exerted to the surrounding medium. The evolutionary changes of its skeleton have been well analyzed in gnathostomes and revealed transitions from heterocercal to diphycercal and homocercal tails. In contrast, we only know little about the evolutionary transformations of the muscular system of the caudalis and about possible ways of force transmission from anterior myomeres to the caudal fin. The goals of this study are to gain insight into evolutionary transformations of the musculoskeletal system in the four basal actinopterygian groups (Cladistia, Chondrostei, Ginglymodi, and Halecomorphi) and to identify likely pathways of force transmission to the tail. In this context, the connective tissue of the myosepta is considered to be an essential part of the musculoskeletal system. For the first time, this system is analyzed for the whole postanal region. The use of microdissection techniques and polarized light microscopy revealed the collagen fiber architecture and the insertions of all postanal myosepta from cleared and stained specimens. The collagen fiber architecture is similar in all investigated specimens and thus represents the primary actinopterygian condition. All parts of postanal myosepta are dominated by longitudinally arranged myoseptal tendons (lateral and myorhabdoid tendons) that span several vertebral segments. This architecture supports the view that posterior myosepta are well designed to transfer muscular forces that are generated in anterior myomeres. In contrast to the uniform myoseptal architecture, the musculoskeletal system differs between the four basal actinopterygian groups. Among them, chondrosteans have retained the plesiomorphic condition of actinopterygian tails. For the remaining taxa several evolutionary novelties in the musculoskeletal system of the tail are revealed. Most of these have evolved independently in the cladistian and neopterygian stem lineage. In these groups extensions of all epaxial and hypaxial parts of myosepta are present that insert on caudal fin rays. This remarkable contribution of epaxial muscle masses to the caudal fin organization is in contrast to the skeletal organization, that largely derives from hypaxial material only. In contrast to former studies the hypochordal longitudinalis muscle is shown to be a synapomorphy of Halecostomi (Halecomorphi + Teleostei). The morphological framework presented here allows to generate new hypotheses on the function of caudal fins that can be tested experimentally.     

Evolutionary transformations of myoseptal tendons in gnathostomes

Axial undulations in fishes are powered by a series of three-dimensionally folded myomeres separated by sheets of connective tissue, the myosepta. Myosepta have been hypothesized to function as transmitters of muscular forces to axial structures during swimming, but the difficulty of studying these delicate complex structures has precluded a more complete understanding of myoseptal mechanics. We have developed a new combination of techniques for visualizing the three-dimensional morphology of myosepta, and here we present their collagen-fibre architecture based on examination of 62 species representing all of the major clades of notochordates. In all gnathostome fishes, each myoseptum bears a set of six specifically arranged tendons. Because these tendons are not present outside the gnathostomes (i.e. they are absent from lampreys, hagfishes and lancelets), they represent evolutionary novelties of the gnathostome ancestor. This arrangement has remained unchanged throughout 400 Myr of gnathostome evolution, changing only on the transition to land. The high uniformity of myoseptal architecture in gnathostome fishes indicates functional significance and may be a key to understanding general principles of fish swimming mechanics. In the design of future experiments or biomechanical models, myosepta have to be regarded as tendons that can distribute forces in specific directions.     

Force transmission via axial tendons in undulating fish: a dynamic analysis

Sonomicrometrics of in vivo axial strain of muscle has shown that the swimming fish body bends like a homogenous, continuous beam in all species except tuna. This simple beam-like behavior is surprising because the underlying tendon structure, muscle structure and behavior are complex. Given this incongruence, our goal was to understand the mechanical role of various myoseptal tendons. We modeled a pumpkinseed sunfish, Lepomis gibbosus, using experimentally-derived physical and mechanical attributes, swimming from rest with steady muscle activity. Axially oriented muscle-tendons, transverse and axial myoseptal tendons, as suggested by current morphological knowledge, interacted to replicate the force and moment distribution. Dynamic stiffness and damping associated with muscle activation, realistic muscle force generation, and force distribution following tendon geometry were incorporated. The vertebral column consisted of 11 rigid vertebrae connected by joints that restricted bending to the lateral plane and endowed the body with its passive viscoelasticity. In reaction to the acceleration of the body in an inviscid fluid and its internal transmission of moment via the vertebral column, the model predicted the kinematic response. Varying only tendon geometry and stiffness, four different simulations were run. Simulations with only intrasegmental tendons produced unstable axial and lateral tail forces and body motions. Only the simulation that included both intra- and intersegmental tendons, muscle-enhanced segment stiffness, and a stiffened caudal joint produced stable and large lateral and axial forces at the tail. Thus this model predicts that axial tendons function within a myomere to (1) convert axial force to moment (moment transduction), (2) transmit axial forces between adjacent myosepta (segment coupling), and, intersegmentally, to (3) distribute axial forces (force entrainment), and (4) stiffen joints in bending (flexural stiffening). The fact that all four functions are needed to produce the most realistic swimming motions suggests that axial tendons are essential to the simple beam-like behavior of fish.     

Morphology and mechanics of myosepta in a swimming salamander (Siren lacertina)

In contrast to the complex, three-dimensional shape of myomeres in teleost fishes, the lateral hypaxial muscles of salamanders are nearly planar and their myosepta run in a roughly straight line from mid-lateral to mid-ventral. We used this relatively simple system as the basis for a mathematical model of segmented musculature. Model results highlight the importance of the mechanics of myosepta in determining the shortening characteristics of a muscle segment. We used sonomicrometry to measure the longitudinal deformation of myomeres and the dorsoventral deformation of myosepta in a swimming salamander (Siren lacertina). Sonomicrometry results show that the myosepta allow some dorsoventral lengthening, indicating an amplification of myomere shortening that is greater than that produced by muscle fiber angle alone (10% muscle fiber shortening produces 28.7% myomere shortening). Polarized light and DIC microscopy of isolated hypaxial myosepta revealed that the collagen fiber orientation in hypaxial myomeres is primarily mediolateral. The mediolateral collagen fiber orientation, combined with our finding that the hypaxial myosepta lengthen dorsoventrally during swimming, suggests that one possible function of hypaxial myosepta in S. lacertina is to increase the strain amplification of the muscle fibers by reducing the mediolateral bulging of the myomeres and redirecting the bulging toward the dorsoventral direction.     

Prey capture in lizards: differences in jaw–neck–forelimb coordination

Fish body muscles are arranged along the vertebral column in three‐dimensional W‐shaped blocks, called myomeres. Each myomere is separated from its neighbours by a collagenous sheet, the myoseptum, and embedded in these myosepta and in positions that are conserved throughout gnathostome evolution are distinct tendons. Within teleosts these tendons often ossify. Ossification is usually intramembranous but cartilaginous structures within the tendons have also been reported. Ossified myoseptal tendons are homologous to intermuscular bones and appear only in teleosts. The phylogenetic signal of myoseptal tendon ossfication has not been tested previously, although the presence and morphology of intermuscular bones have been used to infer phylogenetic relationships. We sample over a broad phylogenetic range of teleost fishes to test for (1) the effects of phylogenetic history on the presence of intermuscular bones and (2) morphological correlations with the presence of intermuscular bones. Body shape and fin position as well as vertebral number and aspect ratio are characters that are likely to affect the distribution of stresses along myoseptal tendons, and are therefore good functional predictors of myoseptal tendon ossification. We use the summary information by Patterson & Johnson for a list of species with intermuscular bones and reanalyse the homology of intermuscular bones to myoseptal tendons. We find that there is a phylogenetic signal in the distribution of four out of six ossified tendons, but that after correcting for phylogenetic relationships there are still morphological predictors for the presence of all ossified tendons. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 607–622.     

The evolution of tendon--morphology and material properties

Phylogenetically, tendinous tissue first appears in the invertebrate chordate Branchiostoma as myosepta. This two-dimensional array of collagen fibers is highly organized, with fibers running along two primary axes. In hagfish the first linear tendons appear and the myosepta have developed specialized regions with unidirectional fiber orientation-a linear tendon within the flat sheet of myoseptum. Tendons react to compressive load by first forming a fibrocartilaginous pad, and under severe stress, sesamoid bones. Evidence for this ability to react to load first arises in the cartilaginous fish, here documented in a tendon from the jaw of a hard-prey crushing stingray. Sesamoid bones are common in bony fish and also in tetrapods. Tendons will also calcify under tensile loads in some groups of birds, and this reaction to load is seen in no other vertebrates. We conclude that the evolutionary history of tendon gives us insight into the use of model systems for investigating tendon biology. Using mammal and fish models may be more appropriate than avian models because of the apparent evolution of a novel reaction to tensile loads in birds.     

Ring bands in fish skeletal muscle: Reorienting the myofibrils and microtubule cytoskeleton within a single cell

Skeletal muscle cells (fibers) contract by shortening their parallel subunits, the myofibrils. Here we show a novel pattern of myofibril orientation in white muscle fibers of large black sea bass, Centropristis striata. Up to 48% of the white fibers in fish >1168 g had peripheral myofibrils undergoing an ～90^o shift in orientation. The resultant ring band wrapped the middle of the muscle fibers and was easily detected with polarized light microscopy. Transmission electron microscopy showed that the reoriented myofibrils shared the cytoplasm with the central longitudinal myofibrils. A microtubule network seen throughout the fibers surrounded nuclei but was mostly parallel to the long‐axis of the myofibrils. In the ring band portion of the fibers the microtubule cytoskeleton also shifted orientation. Sarcolemmal staining with anti‐synapsin was the same in fibers with or without ring bands, suggesting that fibers with ring bands have normal innervation and contractile function. The ring bands appear to be related to body‐mass or age, not fiber size, and also vary along the body, being more frequent at the midpoint of the anteroposterior axis. Similar structures have been reported in different taxa and appear to be associated with hypercontraction of fibers not attached to a rigid structure (bone) or with fibers with unusually weak links between the sarcolemma and cytoskeleton, as in muscular dystrophy. Fish muscle fibers are attached to myosepta, which are flexible and may allow for fibers to hypercontract and thus form ring bands. The consequences of such a ring band pattern might be to restrict the further expansion of the sarcolemma and protect it from further mechanical stress. J. Morphol., 2012. © 2012 Wiley Periodicals, Inc.     

Surgical anatomy of the levator veli palatini: a previously undescribed tendinous insertion of the anterolateral fibers

The purpose of this study was to describe the previously unreported tendinous insertion of the anterolateral fibers of the levator veli palatini (levator) and discuss possible implications for levator function and cleft palate repair. The velopharyngeal anatomy in normal adult cadavers was studied, with histologic confirmation of anatomical findings. These findings were compared with a more limited study of levator anatomy in cleft palates at the time of intraoperative muscle dissection. Just before entering the velum, the levator divides into two parts. The smaller bundle of muscle fibers (anterolateral part) runs anteriorly, close to the lateral pharyngeal wall, and inserts into the palatine aponeurosis through a number of fine tendons. The main part of the muscle runs medially into the velum, where it fans out and forms the levator sling with the contralateral levator. The possible function of the anterolateral part of the levator is discussed. Inadequate release of the tendinous insertions at the time of palate repair may tether the levator anteriorly and compromise muscle retropositioning or may result in splitting of the levator, so that only part of the levator is retropositioned.     

Quantitative microanatomy of jaw muscle attachment in extant diapsids

Muscular reconstructions in vertebrate paleontology have often relied heavily on the presence of "muscle scars" and similar osteological correlates of muscle attachment, a practice complicated by the fact that approximately half of tendinous muscle attachments to bone in extant vertebrates do not leave readily interpretable scars. Microanatomical and histological correlates of tendinous muscle attachment are much less ambiguous. This study examines the microanatomical correlates of muscle attachment for the mandibular adductors in six species of diapsids. Most prominent tendinous or aponeurotic muscle attachments display a high density of extrinsic fibers (similar to Sharpey's fibers). There is also some indication that the density of extrinsic fibers at an attachment may be directly related to the amount of stress exerted on that attachment. The presence of comparable densities of extrinsic fibers in fossil tissue constitutes strong and readily interpretable positive evidence for the presence of adjacent fibrous connective tissue in life. Microanatomy and histology provide reliable data about muscle attachments that cannot be gleaned from gross observation alone. These additional data, when coupled with existing muscular reconstruction techniques, may be essential to the resolution of ambiguous character states, and will provide more severe tests for long-standing hypotheses of musculature in extinct diapsids. Increasing the accuracy and precision of muscular reconstructions lends greater strength to any phylogenetic, paleobiological, or paleoecological inferences that draw upon these reconstructions as important lines of evidence.     

Development of the zebrafish myoseptum with emphasis on the myotendinous junction

Zebrafish myosepta connect two adjacent muscle cells and transmit muscular forces to axial structures during swimming via the myotendinous junction (MTJ). The MTJ establishes transmembrane linkages system consisting of extracellular matrix molecules (ECM) surrounding the basement membrane, cytoskeletal elements anchored to sarcolema, and all intermediate proteins that link ECM to actin filaments. Using a series of zebrafish specimens aged between 24 h post-fertilization and 2 years old, the present paper describes at the transmission electron microscope level the development of extracellular and intracellular elements of the MTJ. The transverse myoseptum development starts during the segmentation period by deposition of sparse and loosely organized collagen fibrils. During the hatching period, a link between actin filaments and sarcolemma is established. The basal lamina underlining sarcolemma is well differentiated. Later, collagen fibrils display an orthogonal orientation and fibroblast-like cells invade the myoseptal stroma. A dense network of collagen fibrils is progressively formed that both anchor myoseptal fibroblasts and sarcolemmal basement membrane. The differentiation of a functional MTJ is achieved when sarcolemma interacts with both cytoskeletal filaments and extracellular components. This solid structural link between contractile apparatus and ECM leads to sarcolemma deformations resulting in the formation of regular invaginations, and allows force transmission during muscle contraction. This paper presents the first ultrastructural atlas of the zebrafish MTJ development, which represents an useful tool to analyse the mechanisms of the myotendinous system formation and their disruption in muscle disorders.