Fast drum strokes: Novel and convergent features of sonic muscle ultrastructure,innervation, and motor neuron organization in the pyramid butterflyfish (hemitaurichthys polylepis)

Sound production that is mediated by intrinsic or extrinsic swim bladder musculature has evolved multiple times in teleost fishes. Sonic muscles must contract rapidly and synchronously to compress the gas‐filled bladder with sufficient velocity to produce sound. Muscle modifications that may promote rapid contraction include small fiber diameter, elaborate sarcoplasmic reticulum (SR), triads at the A–I boundary, and cores of sarcoplasm. The diversity of innervation patterns indicate that sonic muscles have independently evolved from different trunk muscle precursors. The analysis of sonic motor pathways in distantly related fishes is required to determine the relationships between sonic muscle evolution and function in acoustic signaling. We examined the ultrastructure of sonic and adjacent hypaxial muscle fibers and the distribution of sonic motor neurons in the coral reef Pyramid Butterflyfish (Chaetodontidae: Hemitaurichthys polylepis) that produces sound by contraction of extrinsic sonic muscles near the anterior swim bladder. Relative to adjacent hypaxial fibers, sonic muscle fibers were sparsely arranged among the endomysium, smaller in cross‐section, had longer sarcomeres, a more elaborate SR, wider t‐tubules, and more radially arranged myofibrils. Both sonic and non‐sonic muscle fibers possessed triads at the Z‐line, lacked sarcoplasmic cores, and had mitochondria among the myofibrils and concentrated within the peripheral sarcoplasm. Sonic muscles of this derived eutelost possess features convergent with other distant vocal taxa (other euteleosts and non‐euteleosts): small fiber diameter, a well‐developed SR, and radial myofibrils. In contrast with some sonic fishes, however, Pyramid Butterflyfish sonic muscles lack sarcoplasmic cores and A–I triads. Retrograde nerve label experiments show that sonic muscle is innervated by central and ventrolateral motor neurons associated with spinal nerves 1–3. This restricted distribution of sonic motor neurons in the spinal cord differs from many euteleosts and likely reflects the embryological origin of sonic muscles from hypaxial trunk precursors rather than occipital somites. J. Morphol., 2013. © 2012 Wiley Periodicals, Inc.     

Knockdown and overexpression of Unc-45b result in defective myofibril organization in skeletal muscles of zebrafish embryos

Background  

Unc-45 is a myosin chaperone and a Hsp90 co-chaperone that plays a key role in muscle development. Genetic and biochemical studies in C. elegans have demonstrated that Unc-45 facilitates the process of myosin folding and assembly in body wall muscles. Loss or overexpression of Unc-45 in C. elegans results in defective myofibril organization. In the zebrafish Danio rerio, unc-45b, a homolog of C. elegans unc-45, is expressed in both skeletal and cardiac muscles. Earlier studies indicate that mutation or knockdown of unc-45b expression in zebrafish results in a phenotype characterized by a loss of both thick and thin filament organization in skeletal and cardiac muscle. The effects of unc-45b knockdown on other sarcomeric structures and the phenotype of Unc-45b overexpression, however, are poorly understood in vertebrates.     

Structure of the eye of Haideotriton wallacei, a North American troglobitic salamander

The eye of Haideotriton wallacei is more reduced histologically than those of othe troglobitic salamanders. The tiny eye is imbedded in a mass of adipose tissue. No extrinsic eye muscles are present. A rudimentary lens is present in about half of the eyes examined. In two instances the lens is surrounded by a small chamber; most eyes lack a chamber. The retina and iris are relatively undifferentiated. The relatively massive retina lacks rods and cones, an outer plexiform layer and subdivided nuclear layers. A tiny optic nerve runs to the brain.     

Cytomorphometry of sarcoplasmic reticulum in extrinsic eye muscles of the teleost (Tinca tinca L.)

Summary The sarcoplasmic reticulum of the white and red muscle fibers in the extrinsic eye muscle of the tench (Tinca tinca L.) differs in its organization. The SR of the white fibers is organized according to the Z type, the SR of the red fibers according to the A/I type. In order to estimate the volume and surface of the SR component in both types of muscle fibers, a stereological analysis was carried out.The volume of the SR expressed as a volume density (Vv _SR) is twofold greater in the white than in the red muscle fibers. The surface of the SR, expressed as surface density (Sv _SR) is 20,5% larger in the white muscle fibers.I extend my sincerest thanks to Prof. E. R. Weibel for helpful discussion and instructions in the use of his method and to Mrs. Anna Mleczko-Michalik for skillfull technical assistance. — This work was supported by Polish Academy of Science, project number 09.4.1.4.5.     

Coordinated development of myofibrils, sarcoplasmic reticulum and transverse tubules in normal and dysgenic mouse skeletal muscle, in vivo and in vitro

We studied the development of transverse (T)-tubules and sarcoplasmic reticulum (SR) in relationship to myofibrillogenesis in normal and dysgenic (mdg/mdg) mouse skeletal muscle by immunofluorescent labeling of specific membrane and myofibrillar proteins. At E16 the development of the myofibrils and membranes in dysgenic and normal diaphragm was indistinguishable, including well developed myofibrils, a delicate network of T-tubules, and a prominent SR which was not yet cross-striated. In diaphragms of E18 dysgenic mice, both the number and size of muscle fibers and myofibrillar organization were deficient in comparison to normal diaphragms, as previously reported. T-tubule labeling was abnormal, showing only scattered tubules and fragments. However, many muscle fibers displayed cross striation of sarcomeric proteins and SR comparable to normal muscle. In cultured myotubes, cross-striated organization of sarcomeric proteins proceeded essentially in two stages: first around the Z-line and later in the A-band. Sarcomeric organization of the SR coincided with the first stage, while the appearance of T-tubules in the mature transverse orientation occurred infrequently, only after A-band maturation. In culture, myofibrillar and membrane organization was equivalent in normal and dysgenic muscle at the earlier stage of development, but half as many dysgenic myotubes reached the later stage as compared to normal. We conclude that the mdg mutation has little effect on the initial stage of membrane and myofibril development and that the deficiencies often seen at later stages result indirectly from the previously described absence of dihydropyridine receptor function in the mutant.     

Neural activity pattern is not necessary for the development of adult ultrastructure in katydid (Neoconocephalus robustus) singing muscles

Summary The singing muscles of the katydid Neoconocephalus robustus develop adult ultrastructure late in the last nymphal instar and during the first few days of adult life. The ultrastructural changes during early adulthood were not affected by unilateral axotomy shortly after the adult molt. Both denervated and innervated muscles developed adult proportions of mitochondria, myofibril, and sarcoplasmic reticulum and transverse tubules.     

Jaw muscle fiber type distribution in Hawaiian gobioid stream fishes: histochemical correlations with feeding ecology and behavior

Differences in fiber type distribution in the axial muscles of Hawaiian gobioid stream fishes have previously been linked to differences in locomotor performance, behavior, and diet across species. Using ATPase assays, we examined fiber types of the jaw opening sternohyoideus muscle across five species, as well as fiber types of three jaw closing muscles (adductor mandibulae A1, A2, and A3). The jaw muscles of some species of Hawaiian stream gobies contained substantial red fiber components. Some jaw muscles always had greater proportions of white muscle fibers than other jaw muscles, independent of species. In addition, comparing across species, the dietary generalists (Awaous guamensis and Stenogobius hawaiiensis) had a lower proportion of white muscle fibers in all jaw muscles than the dietary specialists (Lentipes concolor, Sicyopterus stimpsoni, and Eleotris sandwicensis). Among Hawaiian stream gobies, generalist diets may favor a wider range of muscle performance, provided by a mix of white and red muscle fibers, than is typical of dietary specialists, which may have a higher proportion of fast-twitch white fibers in jaw muscles to help meet the demands of rapid predatory strikes or feeding in fast-flowing habitats.     

Characterization of the primary sonic muscles in Carapus acus (Carapidae): a multidisciplinary approach

Sound production in carapid fishes results from the action of extrinsic muscles that insert into the swim bladder. Biochemical, histochemical and morphological techniques were used to examine the sonic muscles and compare them with epaxial muscles in Carapus acus. Sonic fibres are thicker than red and thinner than white epaxial fibres, and sonic fibres and myofibrils exhibit an unusual helicoidal organization: the myofibrils of the centre are in a straight line whereas they are more and more twisted towards the periphery. Sonic muscles have both features of red (numerous mitochondria, high glycogen content) and white (alkali-stable ATPase) fibres. They differ also in the isoforms of the light chain (LC3) and heavy chain (HC), in having T tubules at both the Z-line and the A-I junction and in a unique parvalbumin isoform (PAI) that may aid relaxation. All these features lead to the expression of two assumptions about sound generation: the sonic muscle should be able to perform fast and powerful contractions that provoke the forward movement of the forepart of the swim bladder and the stretching and "flapping" of the swim bladder fenestra; the helicoidal organization allows progressive drawing of the swim bladder fenestra which emits a sound when rapidly released in a spring-like manner.     

Expression and Functional Characterization of Smyd1a in Myofibril Organization of Skeletal Muscles

Background

Smyd1, the founding member of the Smyd family including Smyd-1, 2, 3, 4 and 5, is a SET and MYND domain containing protein that plays a key role in myofibril assembly in skeletal and cardiac muscles. Bioinformatic analysis revealed that zebrafish genome contains two highly related smyd1 genes, smyd1a and smyd1b. Although Smyd1b function is well characterized in skeletal and cardiac muscles, the function of Smyd1a is, however, unknown.

Methodology/Principal Findings

To investigate the function of Smyd1a in muscle development, we isolated smyd1a from zebrafish, and characterized its expression and function during muscle development via gene knockdown and transgenic expression approaches. The results showed that smyd1a was strongly expressed in skeletal muscles of zebrafish embryos. Functional analysis revealed that knockdown of smyd1a alone had no significant effect on myofibril assembly in zebrafish skeletal muscles. However, knockdown of smyd1a and smyd1b together resulted in a complete disruption of myofibril organization in skeletal muscles, a phenotype stronger than knockdown of smyd1a or smyd1b alone. Moreover, ectopic expression of zebrafish smyd1a or mouse Smyd1 transgene could rescue the myofibril defects from the smyd1b knockdown in zebrafish embryos.

Conclusion/Significance

Collectively, these data indicate that Smyd1a and Smyd1b share similar biological activity in myofibril assembly in zebrafish embryos. However, Smyd1b appears to play a major role in this process.     

Morphological and histochemical characterization of the pectoral fin muscle of batoids

Batoids differ from other elasmobranch fishes in that they possess dorsoventrally flattened bodies with enlarged muscled pectoral fins. Most batoids also swim using either of two modes of locomotion: undulation or oscillation of the pectoral fins. In other elasmobranchs (e.g., sharks), the main locomotory muscle is located in the axial myotome; in contrast, the main locomotory muscle in batoids is found in the enlarged pectoral fins. The pectoral fin muscles of sharks have a simple structure, confined to the base of the fin; however, little to no data are available on the more complex musculature within the pectoral fins of batoids. Understanding the types of fibers and their arrangement within the pectoral fins may elucidate how batoid fishes are able to utilize such unique swimming modes. In the present study, histochemical methods including succinate dehydrogenase (SDH) and immunofluoresence were used to determine the different fiber types comprising these muscles in three batoid species: Atlantic stingray (Dasyatis sabina), ocellate river stingray (Potamotrygon motoro) and cownose ray (Rhinoptera bonasus). All three species had muscles comprised of two muscle fiber types (slow-red and fast-white). The undulatory species, D. sabina and P. motoro, had a larger proportion of fast-white muscle fibers compared to the oscillatory species, R. bonasus. The muscle fiber sizes were similar between each species, though generally smaller compared to the axial musculature in other elasmobranch fishes. These results suggest that batoid locomotion can be distinguished using muscle fiber type proportions. Undulatory species are more benthic with fast-white fibers allowing them to contract their muscles quickly, as a possible means of escape from potential predators. Oscillatory species are pelagic and are known to migrate long distances with muscles using slow-red fibers to aid in sustained swimming.     

Shape and disposition of clefts,tubules, and sarcoplasmic reticulum in long and short sarcomere fibers of crab and crayfish

Summary The disposition of surface invaginations (clefts, Z and T tubules) and of the sarcoplasmic reticulum has been examined by electron microscopy at three accelerating voltages (100, 200 and 1000 kV) and by phase-contrast light microscopy in crustacean muscles infiltrated by the Golgi stain. In long-sarcomere, tonic type fibers, an extensive system of invaginating clefts has been observed, along with both Z and T tubules. Z and T tubules form interconnections with each other, but only T tubules form specific contacts with the sarcoplasmic reticulum, which in these fibers forms an extended and continuously fenestrated network. In short-sarcomere, phasic type fibers, a ladder-like disposition of an abundant T network is found. Z tubules are absent in these fibers. The sarcoplasmic reticulum forms more frequent junctions with flattened areas of T tubules and with clefts, but has less extensive free surfaces than in the long-sarcomere fibers.We wish to dedicate this paper to the late Graham Hoyle, whose lifetime of work and interest in the study of muscle from a comparative point of view has been an inspiration to us.     

THE ORGANIZATION OF FLIGHT MUSCLE FIBERS IN THE ODONATA

The cytological organization of flight muscle fibers of Odonata has been investigated. These fibers, in representatives of the Zygoptera and Anisoptera, have been compared and found to be similar, except that, in the former, pairs of lamellar fibrils, rather than single fibrils, alternate with the mitochondria. In each instance, in these synchronous muscles, the actin filaments of the myofibrils are found to lie opposite to and midway between pairs of myosin filaments—a configuration previously reported in asynchronous flight muscle fibers. The disposition of the T system and sarcoplasmic reticulum membranes in glutaraldehyde-fixed anisopteran muscle is described in detail: the T system tubules are shown to be radially continuous across the fiber, and are derived as openmouthed invaginations from the surface cell-membrane. The detailed organization of the dyad junctions between these tubules and the adjoining cisternae of the sarcoplasmic reticulum is described. The accessibility of the T system interior to diffusion exchange with the general extracellular milieu has been investigated by studies on the penetration of ferritin into the fiber: molecules of this marker have been found to diffuse solely along the T system tubules, and their presence in the tubule extremities adjoining the centrally placed nuclei confirms the morphological evidence suggesting that these tubules provide open diffusion channels extending across the radius of the fiber. The possible physiological role of these membrane components and their distribution in synchronous muscles of insects and vertebrates and in asynchronous insect flight muscle are discussed.     

The comparative morphology and evolution of the eyes of caecilians (Amphibia,Gymnophiona)

Summary Caecilians (Amphibia, Gymnophiona) have been reported to have vestigial eyes, to lack some or all of the extrinsic eye muscles and their nerves, and to utilize eye muscles and glands, or derivatives of them, to effect movement of the tentacle, a chemosensory structure unique among vertebrates. Morphological evidence indicates that the eye is a functional photoreceptor in virtually all species examined, with an intact retina and optic nerve. The pattern of retention of extrinsic muscles varies. The ontogeny of the eye of Dermophis mexicanus is typical of that of most vertebrates, though components of accommodation never develop. Several taxa are reported in the literature to lack various eye structures; the present study reveals them to be variously present. Evolutionary trends in caecilian eye morphology include the following: (1) the eye is overlain by thicker, often glandular skin, to overlain by bone as well as skin; (2) extrinsic muscles become attenuate, and some to all may be lost; (3) the retina has the typical vertebrate layered organization, to having a reduced cell number, to becoming net-like rather than stratal; (4) the optic nerve is present, becoming attenuate, perhaps represented only by glial cells; (5) the lens is round (aquatic forms, larval and adult) to spheroid; lens crystalline to cellular (retention of the embryonic condition) to amorphous to absent; (6) the vitreous body is reduced or lost; (7) the cornea adheres to the overlying dermis or periosteum; the lens is free to adherent to cornea to adherent to both cornea and retina. Scolecomorphids have the eye pulled out of the socket and embedded in the tentacle under the skin of the upper jaw. This pattern of trends in eye reduction is similar to that observed in other vertebrate lineages that are fossorial or troglobitic.     

EFFECTS OF INTRINSIC AND EXTRINSIC FACTORS ON POPULATION FRAGMENTATION IN THREE SPECIES OF NORTH AMERICAN MINNOWS (TELEOSTEI: CYPRINIDAE)

Geographic patterns of genetic variation (mitochondrial DNA [mtDNA] and allozymes) were used to examine effects of intrinsic characteristics (e.g., vagility, habitat specificity, and reproductive behaviors) and extrinsic factors (e.g., climatic and geological history) on population fragmentation. The three species of cyprinid fishes examined (Tiaroga cobitis, Meda fulgida, and Agosia chrysogaster) occupied similar historical ranges within the lower Colorado River drainage, but differ in intrinsic characteristics conducive to population fragmentation. Relationships among populations were similar across species, reflecting common historical influences, but results indicate the distribution of variation among species is strongly affected by intrinsic characteristics. Variation within two species (T. cobitis and M. fulgida) is subdivided among populations, suggesting little gene flow among rivers. In contrast, similarity of A. chrysogaster populations throughout the Gila River drainage supports the hypothesis that levels of gene flow are high for this species. Levels of mtDNA divergence were much higher than expected for both T. cobitis and A. chrysogaster suggesting long-term isolation of geographic regions. These results indicate that both long-term and short-term extrinsic factors have shaped basic patterns of variation within these fishes; however, the intrinsic characteristics of each species have strongly affected the population genetic structure of these fishes.     

Ontogenetic diet shift of age‐0 year Pacific bluefin tuna Thunnus orientalis

Stomach contents of 437 age‐0 year Pacific bluefin tuna Thunnus orientalis (20·3–59·4 cm fork length, L_F) caught in the Tsushima Current and the Kuroshio regions around Japan were examined to investigate their ontogenetic diet shift. Prey compositions were diverse and different between regions. Although the seasonal growth patterns were different between regions, ontogenetic diet shifts shared a common pattern. In the Tsushima Current region (Sea of Japan), small T. orientalis (20–25 cm L_F) preyed upon small squid (juvenile Enoploteuthis chunii), and larger ones (25–35 cm L_F) gradually shifted their diet to mesopelagic fish (Maurolicus japonicus). In the Kuroshio region (Pacific Ocean), small T. orientalis (20–25 cm L_F) preyed upon small zooplankton (mostly crustacean larvae), and larger ones (25–40 cm L_F) shifted to epipelagic fishes (Etrumeus teres, Sardinops melanostictus and Engraulis japonicus). The observed data suggest that T. orientalis switch to a diet more based on fish prey items, which have more body mass and greater swimming ability than small squid and zooplankton, after they reach a L_F of 25 cm.     

Fine Structure of Cardiac Sarcomeres in the Black Widow Spider Latrodectus mactans

Fine structural characteristics of the cardiac muscle and its sarcomere organization in the black widow spider, Latrodectus mactans were examined using transmission electron microscopy. The arrangement of cardiac muscle fibers was quite similar to that of skeletal muscle fibers, but they branched off at the ends and formed multiple connections with adjacent cells. Each cell contained multiple myofibrils and an extensive dyadic sarcotubular system consisting of sarcoplasmic reticulum and T‐tubules. Thin and thick myofilaments were highly organized in regular repetitive arrays and formed contractile sarcomeres. Each repeating band unit of the sarcomere had three apparent striations, but the H‐zone and M‐lines were not prominent. Myofilaments were arranged into distinct sarcomeres defined by adjacent Z‐lines with relatively short lengths of 2.0 μm to 3.3 μm. Cross sections of the A‐band showed hexagon‐like arrangement of thick filaments, but the orbit of thin filaments around each thick filament was different from that seen in other vertebrates. Although each thick filament was surrounded by 12 thin filaments, the filament ratio of thin and thick myofilaments varied from 3:1 to 5:1 because thin filaments were shared by adjacent thick filaments.     

Anatomy of the extrinsic gut musculature of Gammarus minus (Crustacea: Amphipoda)

Inserting on the buccal and esophageal foregut of Gammarus minus are numerous pairs of serially arranged dorsal dilator muscles, a single pair of lateral muscles, and two pairs of posterior muscles. Muscles of the cardiac stomach include three dorsal sets, a single pair associated with the pterocardiac ossicles, and two pairs inserting on the ventral aspect. A single pair of muscles inserts on the lateral aspect of the pyloric stomach. The extrinsic muscles of the foregut originate from exoskeletal apodemes of the cephalothoracic cuticle, sockets of the mandible, and a maxillary bridge that lies just ventral to the cardiac stomach. The extrinsic musculature of the hindgut is restricted to the rectal region and consists of paired dorsal and ventral series in an X-configuration. A single unpaired muscle inserts on the ventral midline. Extrinsic muscles of the hindgut originate from the integument of the last pleonic segment. The general arrangement of extrinsic gut muscles in G. minus is similar but not identical to that of other amphipods studied. However, the pattern is quite different from that of other malacostracans.     

Organization and fine structure of extraocular muscles in Carassius and Rana

Summary 1. Two types of muscle fibers, red (-slow) and white (-twitch), have been described in the extraocular muscles of Carassius and Rana, respectively. 2. Red and white muscle fibers occupy a definite position in particular eye muscles and occur in almost constant numerical relation. 3. The red fibers in the fish extraocular muscles are supposedly slow. The position of the triads is at the level of the A/I junction, whereas that of the white muscle fibers is at the Z line level. 4. In the frog the extraocular muscles consist of two types of muscle fibers, which have morphological features of slow and fast fibers, respectively, the triads being localized at the Z line level.This work has been supported by the Polish Academy of Sciences.Authors express their thanks to Doc. Dr. J. Kawiak for help in densitometrography.     

Entrance of colloidal ThO2 tracer into the T tubules and longitudinal tubules of the guinea pig heart

Summary Colloidal ThO₂ particles (diameter of 60 Å) were used as electron-opaque markers to trace the intracellular compartments continuous with the bulk interstitial fluid of guinea pig ventricular muscle. Beating and quiescent hearts in a Langendorff preparation were perfused for 30 min with oxygenated Ringer solution containing 1% ThO₂. The hearts were immediately fixed by perfusing with glutaraldehyde solution. The colloidal ThO₂ particles entered into many of the T tubules and into longitudinallyrunning tubules. No differences in distribution of ThO₂ were observed in a heart which was not exposed to ThO₂ until after it was fixed. Tracer did not penetrate into the intercalated disk clefts in the guinea pig hearts and one frog heart used for comparison. Tubular profiles filled with ThO₂ were not seen in frog heart, an observation which confirms the absence of T tubules in this amphibian. It is concluded that, in mammalian cardiac muscle, the lumens of the longitudinal tubules are continuous with the lumens of the T tubules, forming an extensively interconnected T-L tubular system. Hence, every myofibril has close access to a fluid-filled space which is continuous with the interstitial fluid and which may be of similar cationic composition; such an arrangement should facilitate excitation-contraction coupling.Supported by grants from the American Heart Association and from the Public Health Service (HE-11155, HE-05815 and HE-10384). The authors wish to acknowledge the expert technical assistance of Mrs. Jan Redick and to thank Dr. James Smith of Marquette University for the supply of Thorotrast used in these studies.     

THE ULTRASTRUCTURE OF THE CAT MYOCARDIUM : I. Ventricular Papillary Muscle

The ultrastructure of cat papillary muscle was studied with respect to the organization of the contractile material, the structure of the organelles, and the cell junctions. The morphological changes during prolonged work in vitro and some effects of fixation were assessed. The myofilaments are associated in a single coherent bundle extending throughout the fiber cross-section. The absence of discrete "myofibrils" in well preserved cardiac muscle is emphasized. The abundant mitochondria confined in clefts among the myofilaments often have slender prolongations, possibly related to changes in their number or their distribution as energy sources within the contractile mass. The large T tubules that penetrate ventricular cardiac muscle fibers at successive I bands are arranged in rows and are lined with a layer of protein-polysaccharide. Longitudinal connections between T tubules are common. The simple plexiform sarcoplasmic reticulum is continuous across the Z lines, and no circumferential "Z tubules" were identified. Specialized contacts between the reticulum and the sarcolemma are established on the T tubules and the cell periphery via subsarcolemmal saccules or cisterns. At cell junctions, a 20 A gap can be demonstrated between the apposed membranes in those areas commonly interpreted as sites of membrane fusion. In papillary muscles worked in vitro without added substrate, there is a marked depletion of both glycogen and lipid. No morphological evidence for preferential use of glycogen was found.