The membrane systems of the cardiac muscle cell of Tmetonyx cicada O. Fabricius (Crustacea,Amphipoda)

Summary The membrane systems of the cardiac muscle cell of the amphipod Tmetonyx cicada (O. Fabricius) are described. The sarcolemma invaginates and forms a transverse network of tubules at the level of the Z band. Narrow longitudinal tubules branch from the network and connect to another transverse network of tubules at the H band level, where dyadic and triadic junctions are formed with the sarcoplasmic reticulum. Adjacent myofibrils are normally separated by a well developed double layer of the sarcoplasmic reticulum. In areas where the myofibrils closely approach the outer sarcolemma, peripheral couplings have been found at the level of the H band.     

The membrane systems of the cardiac muscle cell of Euchaeta norvegica Boeck (Crustacea,Copepoda)

Summary The membrane systems of the cardiac muscle cell of the copepod Euchaeta norvegica Boeck are described. The heart wall, which is between 0.12 and 1.36 m thick, consists of an epicardium and a single layer of muscle cells. Invaginations of the sarcolemma forming transverse tubules have been found at all levels of the sarcomere with the exception of the H-band level. The longitudinal tubules of the same system are closely associated with the sarcoplasmic reticulum to form interior couplings at the A-I level of the sarcomere. Triadic couplings at the Z band level were not seen in E. norvegica, but peripheral couplings were demonstrated. Nexuses were found in the intercalated discs.     

Fine structure and differentiation of Ascidian muscle. I. Differentiated caudal musculature of Distaplia occidentalis tadpoles

The structure of the caudal muscle in the tadpole larva of the compound ascidian Distaplia occidentalis has been investigated with light and electron microscopy. The two muscle bands are composed of about 1500 flattened cells arranged in longitudinal rows between the epidermis and the notochord. The muscle cells are mononucleate and contain numerous mitochondria, a small Golgi apparatus, lysosomes, proteid-yolk inclusions, and large amounts of glycogen. The myofibrils and sarcoplasmic reticulum are confined to the peripheral sarcoplasm. Myofibrils are discrete along most of their length but branch near the tapered ends of the muscle cell, producing a Felderstruktur. The myofibrils originate and terminate at specialized intercellular junctional complexes. These myomuscular junctions are normal to the primary axes of the myofibrils and resemble the intercalated disks of vertebrate cardiac muscle. The myofibrils insert at the myomuscular junction near the level of a Z-line. Thin filaments (presumably actin) extend from the terminal Z-line and make contact with the sarcolemma. These thin filaments frequently appear to be continuous with filaments in the extracellular junctional space, but other evidence suggests that the extracellular filaments are not myofilaments. A T-system is absent, but numerous peripheral couplings between the sarcolemma and cisternae of the sarcoplasmic reticulum (SR) are present on all cell surfaces. Cisternae coupled to the sarcolemma are continuous with transverse components of SR which encircle the myofibrils at each I-band and H-band. The transverse component over the I-band consists of anastomosing tubules applied as a single layer to the surface of the myofibril. The transverse component over the H-band is also composed of anastomosing tubules, but the myofibrils are invested by a double or triple layer. Two or three tubules of sarcoplasmic reticulum interconnect consecutive transverse components. Each muscle band is surrounded by a thin external lamina. The external lamina does not parallel the irregular cell contours nor does it penetrate the extracellular space between cells. In contracted muscle, the sarcolemmata at the epidermal and notochordal boundaries indent to the level of each Z-line, and peripheral couplings are located at the base of the indentations. The external lamina and basal lamina of the epidermis are displaced toward the indentations. The location, function, and neuromuscular junctions of larval ascidian caudal muscle are similar to vertebrate somatic striated muscle. Other attributes, including the mononucleate condition, transverse myomuscular junctions, prolific gap junctions, active Golgi apparatus, and incomplete nervous innervation are characteristic of vertebrate cardiac muscle cells.     

The membrane systems of the cardiac muscle cell of Cirolana borealis Lilljeborg (Crustacea,Isopoda)

The membrane systems of the cardiac muscle cell of the isopod Cirolana borealis Lilljeborg are described. The sarcolemma invaginates at the level of the Z band, forming transverse tubules. Narrow tubules branch off in a longitudinal direction from these transverse and radially arranged Tz-tubules forming a transverse collar at each A-I level, where dyadic and triadic junctions are formed with the sarcoplasmic reticulum. Two different orientations of the coupling discs have been detected in the supercontracted sarcomere, and this observation has been discussed. Adjacent myofibrils are separated by a double layer of sarcoplasmic reticulum.     

Ultrastructure of the heart of the sturgeon Acipenser stellatus (Chondrostei)

Summary The ultrastructure of atrial and ventricular myocardial cells from Acipenser stellatus is described. The cells of the atrium are more loosely connected than those of the ventricle. Cell contact is by simple intercalated discs and by desmosomes. The cells are flattened, with peripheral myofibrils and a central region of mitochondria and the nucleus. The sarcoplasmic reticulum consists of subsarcolemmal tubules, that frequently extend towards the central mitochondria. Dyads are small and positioned at any sarcomeric level. No T-tubules are present. Specific granules are restricted to the atrial cell, and are sometimes present within the SR tubules.     

The membrane systems of the cardiac muscle cell of Munida tenuimana G. O. Sars (Crustacea,Decapoda)

The membrane systems of the cardiac muscle cell of Munida tenuimana G. O. Sars are described. The sarcolemma invaginates at the Z level, forming tubules. Narrow tubules branch off in a longitudinal direction from these transverse and radially arranged tubules, forming a narrow transverse collar at the H level where dyadic and triadic junctions are formed with the sarcoplasmic reticulum.     

Ultrastructure of atrial and ventricular myocardium in the pike Esox lucius L. and mackerel Scomber scombrus L. (Pisces)

Summary Atrial and ventricular muscle in the pike and mackerel hearts consists of narrow, branching cells. The atrial cells in the two species are similar whereas the ventricular cells differ. The sarcolemma is attached to the Z and M lines of the sarcomere. Intercalated discs are common, and the transverse parts display desmosomes and intermediate junctions. Nexuses are uncommon and only occur in the longitudinal parts of the intercalated discs. The sarcoplasmic reticulum forms a regular hexagonal network on the myofibrillar surface. Subsarcolemmal cisternae form peripheral couplings at the I-A level. Junctional processes are usually inconspicuous, but an electron dense substance is present between the sarcolemma and the Junctional sarcoplasmic reticulum. Specific heart granules are common in atrial cells of both species and in ventricular cells of the pike, but are very scarce in mackerel ventricular muscle.This work was supported by grants from the Norwegian Research Council for Sciences and the Humanities     

The heart Ultrastructure of lepidopleurus asellus (Spengler) and Tonicella marmorea (Fabricius) (Mollusca: Polyplacophora)

Summary The ultrastructure of the auricles, the ostia, and the ventricle of L. asellus and T. marmorea is described. The heart wall consists of an epicardium, a basement membrane, and an inner loose myocardium. The epicardial cells of the auricle are podocytes. The exposed cell body and the branched processes show pedicles. Ventricular epicardium is flat and simple. The slender, unbranched, mononucleated muscle fibres have a peripheral nucleus located midway along the fibre. Mitochondria are peripherally located, leaving the center to longitudinally running thick and thin myofilaments. Dense bodies and attachment plaques make up the Z-material. Sarcomeres and myofibrils are absent, as are transverse tubules and intercalated disks. The sarcoplasmic reticulum consists of peripheral tubules and subsarcolemmal cisternae, some of which radiate, branch, and run between myofilaments. Couplings are lacking. Ventricular fibres in T. marmorea show nexuses and desmosomes; in L. asellus only nexuses. The muscular ostia are tubular, and muscle fibres resemble those of the ventricle; nexuses are detected in T. marmorea and desmosomes in L. asellus. The only nervous elements observed are some nerve processes, structurally similar to those of other molluscs.Supported by grants from the Norwegian Research Council for Science and Humanities     

Ultrastructure and differentiation of ascidian muscle

Summary The larval caudal musculature of the compound ascidian Diplosoma macdonaldi consists of two longitudinal bands of somatic striated muscle. Approximately 800 mononucleate cells, lying in rows between the epidermis and the notochord, constitute each muscle band. Unlike the caudal muscle cells of most other ascidian larvae, the myofibrils and apposed sarcoplasmic reticulum occupy both the cortical and the medullary sarcoplasm.The cross-striated myofibrils converge near the tapered ends of the caudal muscle cell and integrate into a field of myofilaments. The field originates and terminates at intermediate junctions at the transverse cellular boundaries. Close junctions and longitudinal and transverse segments of nonjunctional sarcolemmata flank the intermediate junctions, creating a transverse myomuscular (TMM) complex which superficially resembles the intercalated disk of the vertebrate heart.A perforated sheet of sarcoplasmic reticulum (SR) invests each myofibril. The sheet of SR spans between sarcomeres and is locally undifferentiated in relation to the cross-striations. Two to four saccular cisternae of SR near each sarcomeric Z-line establish interior (dyadic) couplings with an axial analogue of the vertebrate transverse tubular system. The axial tubules are invaginations of the sarcolemma within and adjacent to the intermediate junctions of the TMM complex.The caudal muscle cells of larval ascidians and the somatic striated muscle fibers of lower vertebrates bear similar relationships to the skeletal organs and share similar locomotor functions. At the cellular level, however, the larval ascidian caudal musculature more closely resembles the vertebrate myocardium.This investigation was supported by Developmental Biology Training Grant No. 5-T01-HD00266 from the National Institute of Child Health and Human Development, National Institutes of Health, by National Research Service Award No. 1-F32-GM05259 (M.J.C.) from the National Institute of General Medical Sciences, National Institutes of Health, and by Research Grant No. BMS 7507689 (R.A.C.) from the National Science Foundation. A portion of this study was carried out at the Friday Harbor Laboratories of the University of Washington, and the authors gratefully acknowledge the cooperation and advice extended by the former Director, Dr. Robert L. FernaldResearch facilities were provided in part by Douglas E. Kelly, Professor and Chairman, Department of Anatomy, University of Southern California School of Medicine, Los Angeles, California 90033, USA. The provisions and counsel are warmly acknowledged     

Ultrastructure of the radula protractor of Busycon canaliculatum

Summary The distribution of the sarcoplasmic reticulum and sarcolemmic tubules in the radula protractor muscle of the whelk, Busycon canaliculatum, has been investigated. The sarcoplasmic reticulum consists of an interconnected system of cisternae and tubular channels. The cisternae are closely associated with the sarcolemma. The tubular channels project from the cisternae into the interior of the cell and run parallel to the long axis of the myofilaments. Parallel tubular channels are interconnected with one another by short branches. This finding of an elaborate sarcoplasmic reticulum supports previous physiological work on this smooth muscle which indicated the presence of an intracellular compartmentalization of calcium ions. There is also an extensive system of tubular invaginations of the sarcolemma which we have termed sarcolemmic tubules. These tubules are 600 Å in diameter and about 0.5 microns in length. There is a substructure associated with the leaflet of the tubular membrane bordering the extracellular space. The sarcolemmic tubules penetrate only half a micron from the surface of the cell and interdigitate with the sarcoplasmic reticulum associated with the sarcolemma. Calculations have shown that the surface area of this smooth muscle cell is more than doubled by the presence of sarcolemmic tubules.     

Structures located at the levels of the Z bands in mouse ventricular myocardial cells

Within ventricular myocardial cells of the mouse, the myoplasmic regions located immediately adjacent to the Z lines of the sarcomeres contain a variety of structures. These include: (1) transversely oriented 10 nm (‘intermediate’) filaments that apparently contribute to the cytoskeleton of the myocardial cell; (2) the majority of the transverse elements of the T-axial tubular system; (3) specialized segments of the sarcoplasmic reticulum (SR) that are closely apposed to the sarcolemma or T-axial tubules (junctional SR); (4) ‘extended junctional SR’ (‘corbular SR’) that exists free of association with the cell membrane; (5) ‘Z tubules’ of SR that are intimately apposed to the Z line substance; and (6) leptofibrils. In addition, fasciae adherentes supplant Z lines where myofibrils insert into the transverse borders (intercalated discs) of the cells. The concentration of these myocardial components at the level of the Z lines suggests that a particular specialization of structural and physiological activities exists in the Z-level regions of the myoplasm. In particular, it appears that the combination of intermediate filaments, T tubules, and Z-level SR elements forms a series of parallel planar bodies that extend across each myocardial cell to impart transverse rigidity. The movement and compartmentation of calcium ion (Ca²⁺) would seem especially active near the Z lines of the myofibrils, in view of the preferential location there of Ca²⁺-sequestering myocardial structures such as T tubules, junctional SR, extended junctional SR and Z tubules.     

THE SARCOPLASMIC RETICULUM OF THE BAT CRICOTHYROID MUSCLE

The bat cricothyroid muscle is believed to participate in the production of the short bursts of frequency modulated ultrasound which these animals use as an echolocation device. The evidence seems to indicate that this muscle must be extremely fast acting. It possesses a very well developed sarcoplasmic reticulum, consisting of intercommunicating longitudinal and transverse tubular elements. The transverse elements, situated at the level of the junction between the A and the I bands, are tripartite complexes of tubules called triads, and these are sometimes replaced by more complex structures, the pentads. The intermediate element of the triad appears as a slender continuous tubule, which can be shown to come into close contact with the sarcolemma and also to share with it certain common staining properties. The longitudinal components of the reticulum consist of very numerous tubules which link successive triads to each other and anastomose to form multiple layers of close-meshed reticula in the interfibrillar sarcoplasm. Both the longitudinal and the transverse elements of the sarcoplasmic reticulum form a continuous network across the muscle fiber. It is suggested that the extraordinary development of the sarcoplasmic reticulum in the bat cricothyroid is related to the unusual physiological properties of this muscle.     

The membrane systems of the cardiac muscle cell of Meganychtiphanes norvegica (M. Sars), euphauciacea,crustacea

Summary The membrane systems of cardiac muscle cells of the euphausiacean Meganychtiphanes norvegica are described. Transverse tubules are found both at the Z-band level (T_z-tubules) and at the H-band level (T_h-tubules). Within the sarcomere narrow longitudinal tubules branch off from the T_z-tubules. At the H-band level these tubules expand forming flattened cisternae in dyadic and triadic couplings with the sarcoplasmic reticulum (SR). Adjacent myofibrils are separated by a well developed SR. Modifications of the SR are seen at the H-band level where junctional cisternae are formed.     

THE SARCOPLASMIC RETICULUM AND TRANSVERSE TUBULES OF THE FROG''S SARTORIUS

The sarcoplasmic reticulum of the frog's sartorius muscle was examined by electron microscopy following sequential fixation in glutaraldehyde and osmium tetroxide and embedding in Epon. The earlier results of Porter and Palade on Ambystoma muscle were confirmed in the sartorius. In addition, the transverse tubules were observed to be continuous across the width of the fiber, a set of flat intermediate cisternae was seen to connect the terminal cisternae to the longitudinal tubules in the A band, and the continuous reticulum collar at the center of the A band was found to be perforated by circular and elongated pores (the fenestrated collar). The transverse tubules have a volume about 0.3 per cent of the fiber volume, and a surface area about 7 times the outer cylindrical surface area for a fiber 100 µ in diameter. The terminal cisternae, the intermediate cisternae, and the longitudinal tubules together with the fenestrated collar each have a volume of 4 to 5 per cent of the fiber volume and a surface area 40 to 50 times the outer surface area of a fiber 100 µ in diameter. Some evidence for continuity of the transverse tubules with the fiber surface is presented, but this is thought to be not so convincing as evidence presented by others. The results are discussed in terms of a possible mechanism for a role of the transverse tubules and sarcoplasmic reticulum in excitation-contraction coupling, as suggested by their morphology and a variety of physiological studies. In this scheme, the transverse tubules are thought to be electrically coupled to the terminal cisternae, so that depolarization of the fiber surface spreads inward along the transverse tubules and to the terminal cisternae, initiating the release of a contraction-activating substance.     

Comparative stereology of mouse atria

The left and right atria of the mouse were compared to each other and to the mouse left ventricle using stereologic techniques. The volume fraction (V_v) and surface area per unit cell volume (S_v) of the interior junctional sarcoplasmic reticulum (IJSR), total JSR and extended JSR were greater in the left atrium than in right. The V_v and S_v of the free SR, transverse tubules, and mitochondria were similar in the two atria. It is suggested that the differences in junctional sarcoplasmic reticulum between the atria can be accounted for by a difference in distribution of two types of cells whose anatomy is analogous to working and conducting fibers in the ventricle. The S_v and V_v of the transverse tubules, mitochondria, and all the components of the sarcoplasmic reticulum except for the free SR were greater in the left ventricle than in either atrium. The greater calcium content and sensitivity to extracellular calcium of the atria may explain the greater volume of free SR in the atria as compared to the left ventricle. The S_v of the plasmalemma of the atria and of the S_v of the plasmalemma of the transverse tubules of the left ventricles supports the suggestion of others that there is a constant ratio of surface area to cell volume in cardiac cells.     

A COMPARISON OF THE FINE STRUCTURES OF FROG SLOW AND TWITCH MUSCLE FIBRES

The organisation of the myofibrils and the sarcoplasmic reticulum in frog slow muscle fibres has been compared with that in twitch fibres. It has been found that the filaments have the same length in the two types of fibre, but that there are differences in their packing: (a) in contrast to the regular arrangement of the I filaments near the Z line in twitch fibres, those in slow fibres are irregularly packed right up to their insertion into the Z line; (b) the Z line itself shows no ordered structure in slow fibres; (c) the fine cross-links seen between the A filaments at the M line level in twitch fibres are not present in slow fibres. The sarcoplasmic reticulum in slow fibres consists of two separate networks of tubules. One set of tubules (diameter about 500 to 800 A) is oriented mainly in a longitudinal direction. The tubules of the other network (diameter about 300 A) are oriented either transversely at approximately Z line level or longitudinally, connecting the transverse tubules. Triads are very rarely found, occurring at only every 5th or 6th Z line of each fibril. The central element of these triads is continuous with the thin tubules. Slow fibres from muscles soaked in ferritin-containing solutions contain ferritin particles in the network of thin tubules, the rest of the sarcoplasm remaining free of ferritin.     

The organization of the sarcoplasmic reticulum in teleost ventricular myocardial cells

Summary The morphology of the sarcoplasmic reticulum in myocardial cells of the ventricle of the trout heart is described as the result of an investigation with the electron microscope. The sarcoplasmic reticulum is sparse in distribution compared to that of birds or mammals but shows a fundamentally similar organization. A very loose network of fine tubules is in intimate contact with the myofibrils but with no local modification with respect to the arrangement of myofilaments within the sarcomeres. There is no special association of the sarcoplasmic reticulum with the Z-bands. Some tubules pass to the cell periphery where they expand to form subsarcolemmal cisternae in which electron-dense matter is often seen. The occurrence of the subsarcolemmal cisternae (peripheral couplings) is random and they are not observed in the vicinity of intercalated discs. The sarcoplasmic reticulum is discussed in relation to excitation-contraction coupling in teleost myocardial cells, and in comparison with that of other vertebrates.I am grateful to Professor J. D. Lever for making certain facilities available, and to Mr. P. F. Hire for photographic assistance.     

The ultrastructure of the atrial myocardial cell in three teleostean species

The atrial myocardial cell in three teleostean species Gadiculus thori J. Smith, Melanogrammus aeglefinus (L.) and Onos cimbrius L. is described. The contractile material is located in the periphery of the cell, whereas the nucleus and mitochondria occupy the central part. The tubules of the sarcoplasmic reticulum are of larger diameter at the Z-band levels than elsewhere in the sarcomere. The transverse part of the intercalated disc is composed of alternating desmosomes and intermediate junctions. In the longitudinal part of the disc desmosomes occur, situated in close proximity to Z-bands. Short nexuses were found in the longitudinal part of the disc, usually near the transverse part. In G. thori numerous atrial specific granules were seen. A high number of mitochondrial granules and glycogen granules seems to occur in O. cimbrius. In the atrial myocardial cell of two or six day-old larvae of M. aeglefinus. the contractile material is organized into thin myofibrils, and numerous ribosomes are located in the sarcoplasm.
The results of the present work are compared to those previously described in the teleostean atrium and ventricle. The differences in the heart structure within the gadoid family are discussed from a functional and an evolutionary point of view.     

The fine structure of muscle in a marine turbellarian

Summary The fine structure of the myofibers of Notoplana acticola as studied by electron microscopy indicates that they are composed of thick myofilaments about 200 Å wide with tapering ends and thin myofilaments about 50 Å wide, arranged alongside each other parallel to the long axis of the cell. There is no orderly transverse arrangement of filaments; instead they appear staggered in the fiber. In cross sections 6 to 10 thin filaments form an orbit around one thick filament with possible cross-linkage between the two types of filaments.Dense bodies are associated with the sarcolemma and with the sarcoplasmic reticulum, and appear to serve as attachments for the thin filaments. Dense bodies are compared to elements forming a fragmented Z-disc.Mitochondria, situated in the periphery or the center of fibers, are associated with granules interpreted as glycogen.The sarcoplasmic reticulum consists of: sacs or cisternae in close proximity to the sarcolemma, longitudinal tubular elements between and parallel to the myofilaments, and a tubular network around the filaments. There is no well-defined sarcolemmal-derived transverse tubule system as described in striated muscles. It is hypothesized that in these muscles, the functional equivalent of the T system may be the area of sarcolemma in contact with the cisternae of the sarcoplasmic reticulum.This work was supported by Grant No. GM 10292 from the U. S. Public Health Service to Professor Richard M. Eakin, Department of Zoology at the University of California, Berkeley, USA, where this investigation was conducted during the author's sabbatical leave of absence from the University of Illinois.I wish to thank Professor Eakin for valuable discussions and for his kind hospitality in extending the facilities of his laboratory and the use of the electron microscope to me, and the John Simon Guggenheim Memorial Foundation for the Fellowship which I held during 1964–65.     

Power and control muscles of cicada song: structural and contractile heterogeneity

Sound production in cicadas is powered by a pair of large muscles whose contractions cause buckling of cuticular tymbals and thereby create sound pulses. Sound is modulated by control muscles that alter the stiffness of the tymbals or change the shape of the abdominal resonance chamber. Muscle ultrastructure and contractile properties were characterized for the tymbal muscle and two control muscles, the ventral longitudinal muscle and the tymbal tensor, of the periodical cicada Magicicada septendecim. The tymbal muscle is a fast muscle that is innervated by a single motoraxon. The control muscles are an order of magnitude less massive than the tymbal muscles, but their innervation patterns were considerably more complex. The tensor muscle is innervated by two axons, each of which evokes rather slow twitches, and the ventral muscle is innervated by at least six axons, some of which produce fast and the others slow contractions. Muscle contraction kinetics correlated well with ultrastructure. Fibers of the tymbal muscle and the portions of the ventral muscle thought to be fast were richly supplied with transverse tubules (T-tubules) and sarcoplasmic reticulum (SR); slow portions of the ventral muscle and the tensor muscle had relatively little SR.Abbreviations SR sarcoplasmic reticulum - TTS transverse tubular system - VLM ventral longitudinal muscle