Skeletal muscle myopathy caused by Triton WR-1339

After administration (1 g/kg every other day for a total of five injections) of Triton WR-1339, the tonic, anterior (ALD) and phasic, posterior (PLD) latissimus dorsi muscles of the chicken underwent distinct pathological modifications. Some of the morphological alterations in the muscles paralleled those seen after administration of chloroquine, increased autophagic vacuole formation in the ALD muscle and swelling of the sarcoplasmic reticulum in the PLD muscle, but other changes were unique to Triton WR-1339. These included loss of myofilaments and whole myofibrils, indentation of the sarcolemma as well as increased numbers of ribosomes in the ALD muscle and swelling of the T-tubular system in the PLD muscle. These results are compared with other lysosome mediated pathologies, as well as with other myopathies.     

THE SARCOPLASMIC RETICULUM AND ITS ASSOCIATION WITH THE T SYSTEM IN AN INSECT

The fine structure of the sarcoplasmic reticulum and the transverse tubular system of the femoral muscle of the cockroach, Leucophaea maderae, was studied after prefixation in glutaraldehyde, postfixation in osmium tetroxide, and embedding in Epon. The sarcoplasmic reticulum in this muscle reveals features not previously reported. The sarcoplasmic reticulum is abundant, consisting mainly of a fenestrated envelope which surrounds each myofibril at all levels in the sarcomere. This sarcoplasmic reticulum envelope is continuous transversally as well as longitudinally along the myofibrils. Dyadic junctions are formed by a single T system element which contacts the unfenestrated sarcoplasmic reticulum of adjacent myofibrils in an alternating manner at the ends of the A band. At the dyads, regularly spaced thickenings of the sarcoplasmic reticulum membranes bordering the dyadic spaces are noted. These thickenings, however, do not contact the T tubule membrane. Typical dyadic contacts also are seen between the cell surface membrane and sarcoplasmic reticulum. Z line-like material is seen in contact with the membranes of the cell surface and longitudinal branches of the T systems.     

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.     

Fine structure of the drum muscles of the piranha (serrasalminae,characidae)

Summary The anterior and the posterior drum muscles of the piranha resemble each other in all essential fine structural aspects: myofibrils are slender; sarcomeres are short compared with those of other drum muscles; mitochondria, located in the periphery of the fibers, are numerous and show an irregular internal structure; and the sarcoplasmic reticulum is abundant. Triads appear at the level of the Z lines. The drum muscles have many motor endplates, which, however, lack the characteristic junctional-fold apparatus. No lipid substances could be demonstrated in these muscles. In the posterior drum muscle the fibers depart from their orderly longitudinal arrangement at irregular intervals.     

Ultrastructure of the cyst of Sarcocystis muris.

Cysts of Sarcocystis muris develop within muscle cells and each is bounded by a parasitophorous vacuole membrane. Closely spaced spherical blebs formed from this membrane extend into the muscle cell cytoplasm. A dense substance fills the cavity of the bleb and occupies the vacuolar space immediately adjacent to the membrane. The remainder of the vacuole is filled with a moderately dense matrix within which the parasites develop. At 40 days after infection only metrocytes are present, characterized by their ovoid shape, lightly stained cytoplasm, amylopectin-like granules, and lack of micronemes. Metrocytes divide by a process resembling endodyogeny and eventually produce bradyzoites. By 78 days after infection, at which time the cyst is infective for cats, the few remaining metrocytes are located at the cyst periphery but most organisms are elongated and contain organalles characteristic for bradyzoites including micronemes, dense granules, and amylopectin. Structures indicative of division were not seen in bradyzoites. Rhoptries are few in number. Numerous vesicles of smooth endoplasmic reticulum accumulate in the cytoplasm of muscle cells adjacent to the periphery of the enlarging cyst but significant destruction of muscle fibers containing cysts with viable organisms was not seen in specimens fixed between 40 and 325 days after infection. Unusual lamellar structures were seen in some parasitized muscle cells and intracystic tubules occurred in some cysts.     

Histochemical and Ultrastructural Features of the Biceps Brachii of the African Chameleon (Chamaeleo senegalensis)

Abstract Characteristics of reptilian muscle fibres were investigated in the biceps brachii of the African chameleon, Chamaeleo senegalensis. Fibres were classified as slow and fast. These types of fibre were distinguished on the basis of histochemical staining for myofibrillar ATPase (mATPase). Fast fibres stained dark for mATPase while slow fibres stained light. The patterns of innervation of slow and fast fibres were investigated by staining nerve endings for acetylcholinesterase activity. Slow fibres have a pattern of multiple innervation, whereas fast fibres are associated with individual endplates. The organization of the myofibrils and the sarcoplasmic reticulum in slow muscle fibres from the chameleon biceps brachii was compared with that in fast fibres. Slow fibres lacked an M-line and the Z-lines were uneven. They had fibrils that were not clearly separated from each other and the sarcoplasmic reticulum was poorly developed. These features are in sharp contrast to those of fast fibres which had straight Z-lines, clear M-lines and well-developed sarcoplasmic reticulum.     

Ultrastructure of muscle cells in Siboglinum fiordicum (Pogonophora)

Two different muscle types are found in the body of Siboglinum fiordicum: body wall muscle and blood vessel muscle. Both are of a myomesothelial type. The myofibrils of the body wall muscle are non-striated and consist of thick and thin myofilaments. Scattered dense bodies and attachment plaques are described. The sarcoplasmic reticulum forms a three-dimensional network in the myofibrils and only peripheral couplings are observed. The thick filaments are of a paramyosin type and have a diameter ranging from 400-1500 A. The blood vessels muscle is non-striated, but sometimes a sarcomere-like organization has been observed. Both thick and thin filaments are present. The thick filaments have a diameter of 250-400 A and lack transverse striations. Dense bodies and attachment of plaques are few. The sparse sarcoplasmic reticulum is restricted to the myofibril periphery where it makes peripheral couplings with sarcolemma. The luminal surface of the vessels is lined by a basal lamina with collagen-like inclusions. No endothelium is found. The body wall muscle and the blood vessel muscle are compared with other muscle types described in invertebrates.     

Abnormalities of the nuclear envelope in porcine muscle affected with congenital myofibrillar hypoplasia.

The nuclear ultrastructure of muscle fibres in congenital myofibrillar hypoplasia of piglets was studied in order to provide more information on the pathological changes present. In the initial stages of the disease, muscle fibres with a small reduction of myofibrils often had an increased number of nuclear pores. In the more advanced stages of the disease, the following nuclear abnormalities were observed in muscle fibres showing an extensive disintegration of myofibrils: 1) large dilations of the perinuclear cisternae, 2) small nuclear protrusions projecting into these dilations and 3) appositions of stray membranes of the disrupted endoplasmic reticulum to the nuclear envelope. The increase in the number of nuclear pores may be significant in the development of pathological changes in congenital myofibrillar hypoplasia which is thought to be a hereditary disease. The observed abnormalities of the nuclear envelope appear to be characteristic for muscle fibres undergoing degeneration.     

Ultrastructural stereological analysis of the absolute parameters of the cardiomyocytes in myocardial hypertrophy

Arterial hypertension was produced in male Wistar rats by abdominal aorta ligation. By the 35th day of experiment the animals had developed myocardial hypertrophy. The subcellular organization of cardiomyocytes was studied by electron microscopy and stereology (computation of relative and absolute parameters). In myocardial hypertrophy, the absolute volume of myofibrils, sarcoplasmic reticulum and T system in the left ventricle of the heart was increased, whereas the absolute volume of the mitochondria remained unchanged. The general surface area of all organelles was noticeably increased. The myofibrils and sarcoplasmic reticulum experienced greater changes, which may reflect the features of compensatory processes under the experimental conditions described.     

A comparative study of the organization of the sarcotubular system in ascidian muscle

The caudal musculature of ascidian tadpole larvae consists of mononucleated muscle cells joined end to end in long rows flanking the notochord. A comparative study of the fine structure of these cells in larvae from different families has revealed wide variations in the pattern of organization of the sarcotubular system. The species examined can be distinguished in two groups according to the presence or absence of a system of plasma membrane invaginations equivalent to the T system of vertebrate and invertebrate striated muscle. Muscle cells from the first group of species, Clavelina lepadiformis, Ciona intestinalis and Molgula socialis, are characterized by absence of T system and show peripheral couplings of sarcoplasmic reticulum cisternae directly with the plasma membrane. In contrast, a T system is present in muscle cells of Diplosoma listerianum, Styela plicata and Botrylloides leachi. The presence of T system in ascidian muscle is not related to the taxonomic position of the various species, but rather to the intracellular disposition of the myofibrils, which are peripheral in the species of the first group whereas they occupy a more internal position in the species of the second group. The T system displays unique structural features in ascidian muscle. It consists of wide laminae invaginating from the plasma membrane and associated in longitudinally oriented dyads with sarcoplasmic reticulum cisternae in register with the I band of the myofibrils. It is apparent from these observations that, in contrast with the uniformity of myofibrillar structure in all chordates, there are basic differences between ascidians and vertebrates as regards the organization of the sarcotubular system. On the other hand, there are significant similarities in this respect between ascidian and invertebrate muscle.     

Cold acclimation induces proliferation of sarcoplasmic reticulum without increase in Ca2+-ATPase activity in white axial muscle of striped bass (Morone saxatilis)

The effects of acclimation of striped bass to cold (5 degrees C) and warm (25 degrees C) temperatures upon ultrastructural features of white axial skeletal muscle are quantified. Surface density of sarcoplasmic reticulum (SR) increased by almost 30%, and SR volume density increased by about 20% during cold acclimation. Proliferation of SR suggests an increase in available SR surface for re-sequestration of Ca2+ and a decrease in diffusion path length for Ca2+ during cold acclimation. Average cross-sectional areas and cross-sectional perimeters of myofibrils situated in the center of muscle fibers decreased during cold acclimation by approximately 20% and 11%, respectively. Additionally, average major and minor axes of ellipses fit to central myofibrillar cross-sections decreased by approximately 12% and 8%, respectively, during cold acclimation. These measurements define a decrease in average myofibrillar diameter and suggest a decrease in diffusion path length for Ca2+ to and from myofibrillar activation sites. Measurements of peripheral myofibrils that had elongated profiles in cross-sections indicate that maximum profile length of these myofibrils decreases by about 17%. Peripheral myofibrils may break up into smaller myofibrils with more rounded cross-sectional profiles during cold acclimation. SR Ca2+-ATPase of white axial muscle was also measured in unfractionated homogenates and in crude SR-enriched subcellular fractions from cold- and warm-acclimated striped bass. No difference in SR Ca2+-ATPase activity per g wet weight was observed between cold- and warm-acclimated animals. Lack of increase in SR Ca2+-ATPase per g wet weight, despite a significant proliferation of SR, probably results in a decrease in average Ca2+-ATPase pump density within the SR membrane during cold acclimation. Thus, compensation for decreased diffusion coefficient of Ca2+ during cold acclimation appears due to the combined effects of proliferation of SR surface density and a decrease in average myofibrillar diameter.     

Ultrastructure of gamonts and gametes and fertilization of Sarcocystis sp. from the roe deer (capreolus capreolus) in dogs

Gamonts of Sarcocystis sp. from the roe deer were examined in the intestine of dogs 10 h after inoculation. Early macrogamonts were limited by a three-membranous pellicle, and situated in a parasitophorous vacuole. Female sexual stages during fertilization, the macrogametes, were limited by five membranes, and microgametes were observed in the parasitophorous vacuole. The outer membranes of the microgamete and macrogamete fuse, and the nucleoplasm of the microgamte enters the cytoplasm of the macrogamete. No wall-forming bodies were observed in macrogamonts and macrogametes.     

Penetration of Toxoplasma gondii into host cells induces changes in the distribution of the mitochondria and the endoplasmic reticulum.

Fluorescence microscopy, using dyes which specifically label mitochondria, endoplasmic reticulum and the Golgi complex, and transmission electron microscopy, were used to analyze the changes which occur in the organization of these structures during interaction of Toxoplasma gondii with host cells. In uninfected cells the mitochondria are long filamentous structures which radiate from the nuclear region toward the cell periphery. After parasite penetration they become shorter and tend to concentrate around the parasite-containing vacuole (parasitophorous vacuole) located in the cytoplasm of the host cell. The mitochondria of extracellular parasites, but not of those located within the parasitophorous vacuole, were also stained by rhodamine 123. Labeling with DiOC6, which binds to elements of the endoplasmic reticulum, in association with transmission electron microscopy, revealed a concentration of this structure around the parasitophorous vacuole. The membrane lining this vacuole was also stained, suggesting that components of the endoplasmic reticulum are also incorporated into this membrane. The Golgi complex, as revealed by staining with NBD-ceramide and electron microscopy, maintains its perinuclear position throughout the evolution of the intracellular parasitism.     

Reduction of density and anisotropic distribution of intermediate filaments occur during avian skeletal myogenesis

Chicken skeletal muscle taken from embryos in ovo was examined by thin-section electron microscopy. Measurements of filament diameters reveal three nonoverlapping groups of filaments: thin (actin myofibrillar) filaments with mean diameters of 5.3 +/- 0.6 nm (S.D.), thick (myosin myofibrillar) filaments with mean diameters of 15 +/- 1.4 nm, and intermediate filaments with mean diameters of 9.3 +/- 0.9 nm. During muscle development these diameters do not change. By counting the number of filaments observed in the sarcoplasm at different stages, we find that the spatial density of intermediate filaments decreases during avian myogenesis in ovo, from 91 intermediate filaments/micron 2 at 6 days to 43 intermediate filaments/micron 2 at 17 days in ovo. Initially randomly arranged, some intermediate filaments become associated with Z discs, sarcoplasmic reticulum, nuclear membrane, and the sarcolemma between 6 and 10 days in ovo. These associated intermediate filaments course both parallel and transverse to myofibrils, forming lateral connections between myofibrillar Z discs and longitudinal connections from Z disc to Z disc within myofibrils. Intermediate filaments also appear to connect Z discs with the nuclear membrane. The intermediate filament associations persist through day 17 of development, after which the presence of cytoskeletal filaments is obscured by the densely packed myofibrils and membranes. Intermediate filament distribution becomes anisotropic during development. A greater proportion of intermediate filaments in the immediate perimyofibrillar area are oriented parallel to myofibrils than in other areas, so that the majority of the intermediate filaments nearest the myofibrils course parallel to them. The longitudinal intramyofibrillar intermediate filaments persist throughout development, as shown by their existence in KI-extracted adult myofibrils.     

“Intracisternal microtubules” in neurons of dog ganglia

Summary The changes which occur in the ultrastructure of the mesothoracic retractor unguis muscle of the cockroach Periplaneta americana as a result of disuse are described. Breakdown of myofibrils, sarcoplasmic reticulum, and mitochondria are all marked, this degeneration only being apparent 9 weeks after the operation.     

Ultrastructural stereological analysis of acute myocardial hypertrophy of hypertensive etiology

The myocardium of Wistar rats was studied by electron microscopy after ligation of the renal artery resulting in a stable elevation of blood pressure. The ultrastructural data on the development of heart hypertrophy were described within 5 to 35 days after operation using the morphometric and stereological methods and correlation analysis. The hypertrophied cardiomyocytes showed a reduction in the ratio of the total volume density of the mitochondria, sarcoplasmic reticulum and T system to the volume density of myofibrils. It was discovered that hypertrophy of cardiomyocytes is marked by an increase in the sarcoplasmic reticulum rather than in myofibrils and other cell compartments, whereas the relative volume of mitochondria decreases.     

Studies on the effect of denervation in developing muscle

Summary Ultrastructural diversification of muscle fibers, with regard particularly to myofibrillar changes, has been investigated in the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus muscles of the rat during fetal and postnatal development in the presence and in the absence of motor innervation. The band pattern and the shape of the myofibrils were uniform in fetal and neonatal muscle fibers and underwent differential changes during the first weeks after birth, concomitantly with fiber type specialization. The most evident variations in myofibrillar structure arising in this period concern the thickness of the Z band and the arrangement of the myofibrils. Myofibril formation was at first not impaired by denervation of rat muscles performed in utero and, although focal disintegration of myofibrils and detachment and loss of orientation of filaments became apparent by one week, atrophic muscle fibers with well-organized myofibrils could be seen as late as 2 months after birth. However, denervated muscle fibers of EDL and soleus did not display any significant and consistent difference in myofibrillar band pattern and shape. No variation in mitochondrial content and sarcoplasmic reticulum development was likewise seen in muscle fibers of EDL and soleus after fetal denervation. The findings emphasize the importance of neuromuscular interactions in muscle differentiation.This investigation was supported in part by a grant from Muscular Dystrophy Associations of America, Inc. to Prof. M. Aloisi. A preliminary report of part of this work was presented at the XL Congress of the Italian Zoological Society, Garda, 1971 (Schiaffino, 1972).     

Binding of an ankyrin-1 isoform to obscurin suggests a molecular link between the sarcoplasmic reticulum and myofibrils in striated muscles

Assembly of specialized membrane domains, both of the plasma membrane and of the ER, is necessary for the physiological activity of striated muscle cells. The mechanisms that mediate the structural organization of the sarcoplasmic reticulum with respect to the myofibrils are, however, not known. We report here that ank1.5, a small splice variant of the ank1 gene localized on the sarcoplasmic reticulum membrane, is capable of interacting with a sequence of 25 aa located at the COOH terminus of obscurin. Obscurin is a giant sarcomeric protein of approximately 800 kD that binds to titin and has been proposed to mediate interactions between myofibrils and other cellular structures. The binding sites and the critical aa required in the interaction between ank1.5 and obscurin were characterized using the yeast two-hybrid system, in in vitro pull-down assays and in experiments in heterologous cells. In differentiated skeletal muscle cells, a transfected myc-tagged ank1.5 was found to be selectively restricted near the M line region where it colocalized with endogenous obscurin. The M line localization of ank1.5 required a functional obscurin-binding site, because mutations of this domain resulted in a diffused distribution of the mutant ank1.5 protein in skeletal muscle cells. The interaction between ank1.5 and obscurin represents the first direct evidence of two proteins that may provide a direct link between the sarcoplasmic reticulum and myofibrils.In keeping with the proposed role of obscurin in mediating an interaction with ankyrins and sarcoplasmic reticulum, we have also found that a sequence with homology to the obscurin-binding site of ank1.5 is present in the ank2.2 isoform, which in striated muscles has been also shown to associate with the sarcoplasmic reticulum. Accordingly, a peptide containing the COOH terminus of ank2.2 fused with GST was found to bind to obscurin. Based on reported evidence showing that the COOH terminus of ank2.2 is necessary for the localization of ryanodine receptors and InsP3 receptors in the sarcoplasmic reticulum, we propose that obscurin, through multiple interactions with ank1.5 and ank2.2 isoforms, may assemble a large protein complex that, in addition to a structural function, may play a role in the organization of specific subdomains in the sarcoplasmic reticulum.     

Influence of muscular activity on muscle proteins and sarcoplasmic reticulum content and binding of Ca2+

Training with an increase in intensity of loads causes muscle hypertrophy. The increase of myofibrillar proteins content and proteins of sarcoplasmic reticulum is greater after this training than after training with prolongation of duration of loads. The content of sarcoplasmic proteins is the same in the both kinds of training. The increase in the content of mitochondrial proteins is smaller. The myofibrillar and sarcoplasmic proteins content is the greatest with simultaneous increase in the intensity and duration of loads. Increase in the content of sarcoplasmic vesicles proteins in this case is the same as after training with an increase in the intensity of loads. An increase in the content of mitochondrial proteins is the same as in training with prolongation of duration of loads. The capacity of binding Ca2+ (per unit protein weight), Vmax and Km are not changed. When calculating per unit of muscle mass possibilities of Ca2+ binding under the effect of loads of the uncreasing intensity rise.     

Atypical ‘fibrillar’ flight muscle in strepsiptera

The fine structure of the principal and ancillary metathoracic flight muscle fibres in the adult male of a strepsipteran, Elenchus tenuicornis, is described. Power-producing dorsal longitudinal and dorso-ventral flight muscles show features consistent with myoneural asynchrony: myofibrils are large and discrete and are separated by large closely packed mitochondria; the sarcoplasmic reticulum is very reduced but engages with T-system membranes in dyads at the mid-sarcomere H-band level. With respect to other asynchronous insect flight muscles, the fibres of Elenchus are anomalous (i) in the small fibre diameter, (ii) in the variable contour of the myofibrils and (iii) in the absence of tracheolar invagination. The functional significance of these structural features is discussed. Ancillary metathoracic muscles are structurally comparable with other synchronous fibres in possessing an extensive SR compartment. Structural evidence for asynchrony in the flight mechanism of Strepsiptera is considered in the context of the evolution of this mechanism throughout the insect Orders.