Myofibril Architecture Research Articles

Neddylation drives myofibrillogenesis in the developing heart.

Neddylation is a highly conserved post-translational modification that plays critical roles in various cellular processes through the modulation of cullins and non-cullin substrates. While neddylation is known to be essential for embryonic development, tumor growth, and organogenesis of different tissues, its role in cardiogenesis remains unexplored. Here, we investigated the role of neddylation in early cardiac development by deleting the gene encoding a regulatory subunit of the NEDD8-specific E1 activating enzyme, Nae1, globally and in a heart-specific fashion via Nkx2-5Cre. Global deletion of Nae1 in mice led to embryonic lethality before embryonic day (E) 8.5, whereas cardiac-specific NAE1 knockout mice died at around E12.5 with pronounced cardiac effusion and peripheral hemorrhage, characteristic of cardiac failure. Histological analysis revealed significant thinning of the compact myocardium and reduced trabeculae in mutant hearts, which were accompanied by reduced cardiomyocyte proliferation. Unbiased transcriptomic analysis identified perturbations in cardiomyocyte proliferation and myofibril architecture in mutant hearts. Subsequent analysis showed that loss of NAE1 disrupted sarcomere assembly dysregulated the expression of several important contractile proteins, and impaired mitochondrial function in the developing heart, which was accompanied by downregulation of key cardiac transcription factors including NKX2-5 and SRF. Collectively, our findings demonstrate the essential role of neddylation in cardiogenesis at least in part by driving cardiomyocyte proliferation and myofibrillogenesis.

Myofibril architecture in prediction and evaluation of broiler wooden breast: Histopathological and immunohistochemical study

Myofibril architecture in prediction and evaluation of broiler wooden breast: Histopathological and immunohistochemical study

A new organisational design in skeletal muscle fibres.

In vertebrate skeletal muscles, the architecture of myofibrils is particularly well conserved throughout the taxa. It is composed of suites of repeating functional units called sarcomeres which give the muscle its striated structure. Here, we show that the skeletal sound producing muscles of the cusk eel Parophidion vassali have a different organisation, distinct from the classical type found in textbooks. Within sarcomeres, filaments are not straight lines but have a Y-shaped structure. This looks like chicken wire, with one branch connecting to a branch from the myofibril above and the other connecting to a branch from the myofibril below. This organisation seems to be an adaptation to counteract a trade-off between the speed and force. The low ratio of myofibrils within cell muscles and the high volume of sarcoplasmic reticulum strongly suggest that these muscles are capable of fast contractions. In parallel, the Z-bands are quite wide about 30% of the sarcomere length. This extraordinary long Z-band could smooth out the tension variations found in high-speed muscle contraction, helping to produce sounds with low variabilities in the sound features. Simultaneously, the Y-shaped structure allows having more cross-bridges, increasing the force in this high-speed muscle.

Superior preservation of capillaries, myofibrils and mitochondria after long-term extracorporeal perfusion of free muscle flaps - A descriptive electron microscopy study.

Extracorporeal perfusion (ECP) is a promising technique for prolonged tissue preservation, but might have side effects. For instance, increased radical oxygen species or capillary endothelial damage. To assess ultra-morphological muscle damage during 36-hour ECP of porcine musculocutaneous flaps, hypothesizing that it would delay the onset of damage compared to static cold storage (SCS). Bilateral flaps were retrieved from three Dutch Landrace pigs. Three flaps were preserved for 36 hours by hypothermic storage 4-6°C (control group) and three flaps by ECP with cooled University of Wisconsin solution. Muscle biopsies were taken at 0 h, 12 h and 36 h and assessed with transmission electron microscopy. Muscle architecture was best preserved by ECP, with a delayed onset and decreased severity of muscle damage. After 36 hours, damage was two-fold lower in ECP-flaps compared to SCS-flaps. Myofibril architecture was best preserved. Mitochondria were greatly preserved with swelling being the most prominent feature. Capillaries were moderately but differently damaged during ECP, with focal endothelial thinning as opposed to luminal obstruction in SCS-preserved flaps. This experiment described favourable cellular preservation of skeletal muscle flaps during ECP compared to SCS. Results showed less severe ultra-morphological damage and a later onset of damage.

The insect perspective on Z-disc structure and biology.

Myofibrils are the intracellular structures formed by actin and myosin filaments. They are paracrystalline contractile cables with unusually well-defined dimensions. The sliding of actin past myosin filaments powers contractions, and the entire system is held in place by a structure called the Z-disc, which anchors the actin filaments. Myosin filaments, in turn, are anchored to another structure called the M-line. Most of the complex architecture of myofibrils can be reduced to studying the Z-disc, and recently, important advances regarding the arrangement and function of Z-discs in insects have been published. On a very small scale, we have detailed protein structure information. At the medium scale, we have cryo-electron microscopy maps, super-resolution microscopy and protein-protein interaction networks, while at the functional scale, phenotypic data are available from precise genetic manipulations. All these data aim to answer how the Z-disc works and how it is assembled. Here, we summarize recent data from insects and explore how it fits into our view of the Z-disc, myofibrils and, ultimately, muscles.

Myosin Transducer Inter-Strand Communication Is Critical for Normal ATPase Activity and Myofibril Structure.

Simple SummaryMyosin is a molecular motor protein that is critical for using stored energy to yield contraction in both skeletal muscle and in the heart. Mutations in myosin can result in inherited human hypertrophic cardiomyopathy, wherein the heart walls become thickened. This results in inability to adequately pump enough oxygenated blood to meet bodily demands, and can lead to abnormal heart beating patterns and the possibility of sudden death. Typically, over-active myosin can be causative for this disease. Here, we used the common fruit fly to study one of these cardiomyopathy-causing mutations by expressing the mutant protein in the indirect flight muscle of the fly. Surprisingly, we found that the mutation and associated mutations decrease myosin and muscle function instead of enhancing it. We show that at least some of these defects result from improper interactions within the myosin protein. This suggests that appropriate communications within myosin are critical for its function and further indicates that certain aspects of indirect flight muscle myosin are functionally distinct from those of myosin in the human heart. By understanding the intricate interactions within the myosin motor protein that are important for its normal role, therapeutics for improving mutant myosin function can be developed.The R249Q mutation in human β-cardiac myosin results in hypertrophic cardiomyopathy. We previously showed that inserting this mutation into Drosophila melanogaster indirect flight muscle myosin yields mechanical and locomotory defects. Here, we use transgenic Drosophila mutants to demonstrate that residue R249 serves as a critical communication link within myosin that controls both ATPase activity and myofibril integrity. R249 is located on a β-strand of the central transducer of myosin, and our molecular modeling shows that it interacts via a salt bridge with D262 on the adjacent β-strand. We find that disrupting this interaction via R249Q, R249D or D262R mutations reduces basal and actin-activated ATPase activity, actin in vitro motility and flight muscle function. Further, the R249D mutation dramatically affects myofibril assembly, yielding abnormalities in sarcomere lengths, increased Z-line thickness and split myofibrils. These defects are exacerbated during aging. Re-establishing the β-strand interaction via a R249D/D262R double mutation restores both basal ATPase activity and myofibril assembly, indicating that these properties are dependent upon transducer inter-strand communication. Thus, the transducer plays an important role in myosin function and myofibril architecture.

Correlative imaging of the spatio-angular dynamics of biological systems with multimodal instant polarization microscope.

The spatial and angular organization of biological macromolecules is a key determinant, as well as informative readout, of their function. Correlative imaging of the dynamic spatio-angular architecture of cells and organelles is valuable, but remains challenging with current methods. Correlative imaging of spatio-angular dynamics requires fast polarization-, depth-, and wavelength-diverse measurement of intrinsic optical properties and fluorescent labels. We report a multimodal instant polarization microscope (miPolScope) that combines a broadband polarization-resolved detector, automation, and reconstruction algorithms to enable label-free imaging of phase, retardance, and orientation, multiplexed with fluorescence imaging of concentration, anisotropy, and orientation of molecules at diffraction-limited resolution and high speed. miPolScope enabled multimodal imaging of myofibril architecture and contractile activity of beating cardiomyocytes, cell and organelle architecture of live HEK293T and U2OS cells, and density and anisotropy of white and grey matter of mouse brain tissue across the visible spectrum. We anticipate these developments in joint quantitative imaging of density and anisotropy to enable new studies in tissue pathology, mechanobiology, and imaging-based screens.

Effect of myofibril architecture on the active contraction of dystrophic muscle. A mathematical model

Duchenne muscular dystrophy (DMD) is a muscle degenerative disease caused by a mutation in the dystrophin gene. The lack of dystrophin leads to persistent inflammation, degeneration/regeneration cycles of muscle fibers, Ca2＋ dysregulation, incompletely regenerated fibers, necrosis, fibrotic tissue replacement, and alterations in the fiber ultrastructure i.e., myofibril misalignment and branched fibers. This work aims to develop a comprehensive chemo-mechanical model of muscle-skeletal tissue accounting for dispersion in myofibrillar orientations, in addition to the disorders in sarcomere pattern and the fiber branching. The model results confirm a significant correlation between the myofibrillar dispersion and the reduction of isometric force in the dystrophic muscle and indicate that the reduction of contraction velocity in the dystrophic muscle seems to be associated with the local disorders in the sarcomere patterns of the myofibrils. Also, the implemented model can predict the force–velocity response to both concentric and eccentric loading. The resulting model represents an original approach to account for defects in the muscle ultrastructure caused by pathologies as DMD.

Structural consequences of mutations associated with idiopathic restrictive cardiomyopathy.

Idiopathic restrictive cardiomyopathy (RCM, MIM# 115210) is the least common type of cardiomyopathies, often of genetic origin. Recently we described a spectrum of variants-classified as pathogenic, likely pathogenic and variants of unknown significance-in 24 patients suffering from idiopathic RCM. Pathogenic variants, detected in half of the RCM cases, were found in sarcomeric and cytoskeletal genes that have a predominant role in the development of RCM. Here we have analyzed the structural consequences of these missense variants and predicted their effect on the function of three large groups of domains: intrinsically disordered regions (IDRs), fibronectin-type III (FnIII) domains, and immunoglobulin-like (Ig) domains. Our findings indicate that pathogenic mutations are likely to disrupt interdomain interfaces, interfere with protein interactions, and affect protein stability, potentially destabilizing the multi-domain architecture of myofibrils and leading to myocardial stiffness in patients with idiopathic RCM.

Mitochondrial function in engineered cardiac tissues is regulated by extracellular matrix elasticity and tissue alignment.

Mitochondria in cardiac myocytes are critical for generating ATP to meet the high metabolic demands associated with sarcomere shortening. Distinct remodeling of mitochondrial structure and function occur in cardiac myocytes in both developmental and pathological settings. However, the factors that underlie these changes are poorly understood. Because remodeling of tissue architecture and extracellular matrix (ECM) elasticity are also hallmarks of ventricular development and disease, we hypothesize that these environmental factors regulate mitochondrial function in cardiac myocytes. To test this, we developed a new procedure to transfer tunable polydimethylsiloxane disks microcontact-printed with fibronectin into cell culture microplates. We cultured Sprague-Dawley neonatal rat ventricular myocytes within the wells, which consistently formed tissues following the printed fibronectin, and measured oxygen consumption rate using a Seahorse extracellular flux analyzer. Our data indicate that parameters associated with baseline metabolism are predominantly regulated by ECM elasticity, whereas the ability of tissues to adapt to metabolic stress is regulated by both ECM elasticity and tissue alignment. Furthermore, bioenergetic health index, which reflects both the positive and negative aspects of oxygen consumption, was highest in aligned tissues on the most rigid substrate, suggesting that overall mitochondrial function is regulated by both ECM elasticity and tissue alignment. Our results demonstrate that mitochondrial function is regulated by both ECM elasticity and myofibril architecture in cardiac myocytes. This provides novel insight into how extracellular cues impact mitochondrial function in the context of cardiac development and disease.NEW & NOTEWORTHY A new methodology has been developed to measure O2 consumption rates in engineered cardiac tissues with independent control over tissue alignment and matrix elasticity. This led to the findings that matrix elasticity regulates basal mitochondrial function, whereas both matrix elasticity and tissue alignment regulate mitochondrial stress responses.

Design, synthesis, docking and QSAR study of substituted benzimidazole linked oxadiazole as cytotoxic agents, EGFR and erbB2 receptor inhibitors

The synthesis of benzimidazole linked oxadiazole derivatives designed as potential EGFR and erbB2 receptor inhibitors with anticancer and apoptotic activity were studied. Compounds 7a specifically inhibit EGFR and erbB2 receptor at 0.081 and 0.098 μM concentration. Some of the compounds showed strong, broad-spectrum antiproliferative activitiy when tested against five human cancer cell lines. Compounds 7a and 7n were more cytotoxic than 5-fluorouracil against MCF-7 cancer cell, with IC50 values of 5.0 and 2.55 μM whereas, only 7a led to cell cycle arrest at G2/M phase accompanied by an increase in apoptosis. Compounds 7a and 7n showed normal architecture of myofibrils in cardiomyopathy study whereas only compound 7a showed nearly equal biochemical parameters (SGOT and SGPT) when compared to control. Molecular docking & 3D-QSAR studies were used to establish interactions of 7a and 7n within the active site of enzyme for ATP binding site of kinase domain.

Actin binding GFP allows 4D in vivo imaging of myofilament dynamics in the zebrafish heart and the identification of Erbb2 signaling as a remodeling factor of myofibril architecture.

Dilated cardiomyopathy is a leading cause of congestive heart failure and a debilitating complication of antineoplastic therapies. Despite disparate causes for dilated cardiomyopathy, maladaptive cardiac remodeling and decreased systolic function are common clinical consequences, begging an investigation of in vivo contractile dynamics in development and disease, one that has been impossible to date. To image myocardial contractile filament dynamics in vivo and to assess potential causes of dilated cardiomyopathy in antineoplastic therapies targeting the epidermal growth factor receptor Erbb2. We generated a transgenic zebrafish line expressing an actin-binding green fluorescent protein in cardiomyocytes, allowing an in vivo imaging of myofilaments. Analysis of this line revealed architectural differences in myofibrils of the distinct cardiomyocyte subtypes. We used this model to investigate the effects of Erbb2 signaling on myofibrillar organization because drugs targeting ERBB2 (HER2/NEU) signaling, a mainstay of breast cancer chemotherapy, cause dilated cardiomyopathy in many patients. High-resolution in vivo imaging revealed that Erbb2 signaling regulates a switch between a dense apical network of filamentous myofibrils and the assembly of basally localized myofibrils in ventricular cardiomyocytes. Using this novel line, we compiled a reference for myofibrillar microarchitecture among myocardial subtypes in vivo and at different developmental stages, establishing this model as a tool to analyze in vivo cardiomyocyte contractility and remodeling for a broad range of cardiovascular questions. Furthermore, we applied this model to study Erbb2 signaling in cardiomyopathy. We show a direct link between Erbb2 activity and remodeling of myofibrils, revealing an unexpected mechanism with potentially important implications for prevention and treatment of cardiomyopathy.

Reduced muscle fiber force production and disrupted myofibril architecture in patients with chronic rotator cuff tears

A persistent atrophy of muscle fibers and an accumulation of fat, collectively referred to as fatty degeneration, commonly occur in patients with chronic rotator cuff tears. The etiology of fatty degeneration and function of the residual rotator cuff musculature have not been well characterized in humans. We hypothesized that muscles from patients with chronic rotator cuff tears have reduced muscle fiber force production, disordered myofibrils, and an accumulation of fat vacuoles. The contractility of muscle fibers from biopsy specimens of supraspinatus muscles of 13 patients with chronic full-thickness posterosuperior rotator cuff tears was measured and compared with data from healthy vastus lateralis muscle fibers. Correlations between muscle fiber contractility, American Shoulder and Elbow Surgeons (ASES) scores, and tear size were analyzed. Histology and electron microscopy were also performed. Torn supraspinatus muscles had a 30% reduction in maximum isometric force production and a 29% reduction in normalized force compared with controls. Normalized supraspinatus fiber force positively correlated with ASES score and negatively correlated with tear size. Disordered sarcomeres were noted, along with an accumulation of lipid-laden macrophages in the extracellular matrix surrounding supraspinatus muscle fibers. Patients with chronic supraspinatus tears have significant reductions in muscle fiber force production. Force production also correlates with ASES scores and tear size. The structural and functional muscle dysfunction of the residual muscle fibers is independent of the additional area taken up by fibrotic tissue. This work may help establish future therapies to restore muscle function after the repair of chronically torn rotator cuff muscles.

Patients with chronic myosteatosis have a reduction in muscle fiber force production and disrupted myofibril architecture (1163.18)

A persistent atrophy of muscle fibers and an accumulation of fat, collectively referred to as myosteatosis, commonly occurs in patients with chronic muscle tears. The rotator cuff muscle group develops substantial myosteatosis after chronic tears, and patients with these injuries often have marked reductions in upper extremity function and quality of life. As the etiology of myosteatosis has not been well characterized in humans, our objective was to determine changes in muscle fiber structure and function in patients with chronic rotator cuff tears. We hypothesized that muscles from patients with chronic rotator cuff tears have reduced muscle fiber force production, disordered myofibrils and an accumulation of inflammatory macrophages.Compared with control muscles, there was no difference in muscle fiber cross‐sectional area, but torn supraspinatus muscles had a 30% reduction in maximum isometric force production and a 29% reduction in normalized force. Using electron microscopy and histology, disordered sarcomeres were noted, along with an accumulation of lipid‐laden macrophages in the extracellular matrix surrounding muscle fibers. The size of the tear correlated well (R2=0.49, P<0.01) with reductions in force production. The results of the study indicate that patients with myosteatosis have marked reductions in muscle fiber force production, disrupted myofibril architecture and an accumulation of inflammatory macrophages in the muscle extracellular matrix. This work will help to establish future therapies to restore muscle function for patients with myosteatosis.

Reduction in Muscle Fiber Force Production, Disruption of Muscle Cell Architecture and Accumulation of Fatty Macrophages in Patients with Chronic Rotator Cuff Tears

Objectives:A common pathophysiological change that occurs in torn rotator cuff muscles is atrophy of muscle fibers and an accumulation of intramuscular fat, collectively referred to as "fatty degeneration." For many patients with chronic rotator cuff tears that undergo surgical repair, fatty degeneration is not resolved. The etiology of fatty degeneration has not been characterized in humans, and gaining greater insight into the mechanisms that lead to the development of atrophy and fat accumulation will likely improve the recovery of patients who undergo surgical repair of a torn rotator cuff. The purpose of this study was to gain a greater understanding of the changes in muscle fiber contractility, myofibril architecture and fat accumulation in patients with rotator cuff tears. We hypothesized that torn rotator cuff muscles have reduced muscle fiber force production, disordered myofibrils and an accumulation of fat vacuoles and lysosomes.Methods:This study was approved by our institution's IRB. Prior to enrollment in the study, informed consent was obtained from patients with a full thickness supraspinatus tear as verified by MRI or ultrasound by a fellowship-trained musculoskeletal radiologist. Biopsies of the supraspinatus and anterior deltoid were obtained at the time of repair and prepared for muscle contractility testing or microscopy. The contractility of permeabilized muscle fibers was performed as previously described, and force values of torn supraspinatus and intact deltoid muscles were compared to age-matched force values of healthy fibers sampled from the vastus lateralis muscle in a previous study. Scanning electron micrographs and immunohistochemistry were also performed on select muscle biopsies.Results:Compared with healthy muscle fibers from the vastus lateralis, there was a 29% reduction in specific force (maximum isometric force normalized to fiber cross-sectional area) in torn supraspinatus muscles, and a 34% reduction in specific force for deltoid muscle fibers (Figure 1A, P<0.05). Numerous large intramuscular lipid vacuoles and lysosomes were observed in areas of myofibril degradation (Figure 1B) and streaming of force transmitting Z-disks was often present (Figure 1C). Fatty macrophages, also known as foam cells, were also observed in the ECM between muscle fibers in electron micrographs (Figure 1D) and using a macrophage/foam cell surface marker and BODIPY fluorescent lipid stain (Figure 1E).Conclusion:Combined, these results identify chronic structural and mechanical changes in torn rotator cuff muscles that may explain the poor functional capacity of the muscles after repair. While the supraspinatus muscle had reduced force production, a reduction in force was also noted in the deltoid which may parallel symptomatic rotator cuff disease. Further, the accumulation of fat in large lipid vacuoles and high lysosome densities near myofibrils suggest that a portion of the intramyocellular fat that accumulates in torn cuff muscles likely comes as a result of myofibril degradation. We have also identified for the first time that a portion of the extramyocellular fat in torn rotator cuff muscles accumulates in fatty macrophages/foam cells. Future therapies that target lysosomal-mediated myofibril degradation or macrophage accumulation may help to ameliorate the severity of fatty degeneration in patients with rotator cuff tears.

Characterization of Functional Abnormalities in Hearts of Transgenic Mice Expressing Forms of Actin Capping Protein Defective in Attaching Actin Filaments to Z‐lines

In striated muscle, the barbed ends of actin filaments are attached to Z lines. Biochemical and cell biological studies suggest that actin capping protein (CP) mediates this attachment by binding the barbed ends of actin filaments and alpha-actinin crosslinks overlapping filaments from adjacent sarcomeres. Previous transgenic studies revealed that defective interaction between CP and actin filaments causes major structural defects in sarcomere organization, leading to cardiac hypertrophy and lethality. To determine the basis of the myofibril defect, we have examined the hearts of transgenic mice using histological and immunofluorescence microscopy. The hearts of transgenic mice were hypertrophied relative to body size, with an enlargement of the ventricle wall. Immunofluorescence studies revealed an altered actin organization with disruption of myofibril architecture. To understand how this defect affects cardiac function, blood pressure was measured using an indirect tail pressure assay and ECG measurements determined. The transgenic mice had a lower systolic pressure and an increased heart rate compared to wildtype mice. In ECG studies, a significant increase for both the PR and R wave height was detected, indicative of atrial and ventricular hypertrophy, validating histological studies. This research was funded by a Faculty Development Grant, Minnesota State University-Mankato, MN.

Vertebrate isoforms of actin capping protein beta have distinct functions In vivo.

Actin capping protein (CP) binds barbed ends of actin filaments to regulate actin assembly. CP is an alpha/beta heterodimer. Vertebrates have conserved isoforms of each subunit. Muscle cells contain two beta isoforms. beta1 is at the Z-line; beta2 is at the intercalated disc and cell periphery in general. To investigate the functions of the isoforms, we replaced one isoform with another using expression in hearts of transgenic mice. Mice expressing beta2 had a severe phenotype with juvenile lethality. Myofibril architecture was severely disrupted. The beta2 did not localize to the Z-line. Therefore, beta1 has a distinct function that includes interactions at the Z-line. Mice expressing beta1 showed altered morphology of the intercalated disc, without the lethality or myofibril disruption of the beta2-expressing mice. The in vivo function of CP is presumed to involve binding barbed ends of actin filaments. To test this hypothesis, we expressed a beta1 mutant that poorly binds actin. These mice showed both myofibril disruption and intercalated disc remodeling, as predicted. Therefore, CPbeta1 and CPbeta2 each have a distinct function that cannot be provided by the other isoform. CPbeta1 attaches actin filaments to the Z-line, and CPbeta2 organizes the actin at the intercalated discs.

Analysis of Myofibrillar Organization and Degeneration by Fluorescence Confocal Microscopy

Myofibrillar degeneration is an important pathological process in progressive cardiomyopathy leading to heart failure. Loss of myofibrils in vivo has been observed in both adaptive cardiac responses (i.e. hypertrophy) as well as in chemotherapeutic use of antitumor drugs with cardiotoxic side effects (i.e. doxorubicin). The molecular mechanism(s) of myofibrillar degeneration are poorly understood in comparison with the sequence of events involved in myofibrillar assembly and organization. Maintenance of myofibril integrity is dependent upon a variety of factors, including contractile protein stoichiometry and protein kinase activity.The repeating sarcomeric architecture of myofibrils is well suited to structural analysis, since disruption of normal organization is easily visualized by fluorescence microscopy. Many antibodies are available for use in observation of myofibril structure (see Table at right). Confocal microscopy provides advantages in studies of myofilament organization by allowing for direct and accurate measurement of distances to within 0.2 βm and the ability to perform vertical sectioning through individual cells. This vertical (Z-axis) sectioning can be used to select the focal plane within the cell for observation, resulting in higher resolution images by reducing fluorescent signals from above and below the plane. Image analysis software enables the user to create projections of optically sectioned cardiomyocytes which can be rotated to reveal interior structural relationships previously unobserved with single sections or conventional epifluorescence microscopy. Examples of image analysis will highlight useful features such as distance measurement, periodicity, pixel intensity, colocalization of dual labels, and three dimensional reconstruction.

Estudio ultraestructural de la mionecrosis inducida en raton por el veneno de terciopelo ( Bothrops asper) de Costa Rica

Skeletal muscle from white mice was examined one and three hr after i.m. injection of 50 μg of Bothrops asper venom. At the light microscopic level we observed nectoric fibers with myolitic appearance, as early as the first hr and more marked by the third hr. At the ultrastructural level three kinds of fibers were observed: (a) fibers with normal morphology, (b) fibers with slight alteration at the sarcoplasmic reticulum and (c) fibers with strong alterations in several organelles; in these fibers, the main pathological feature was the disruption of myofibril architecture. This disruption started with the disappearance of A, H, I and M bands, followed by the alteration of Z line. At the end of this process an amorphous mass of proteins was observed. Shortly following envenomation, sarcoplasmic reticulum showed moderate dilatation and proliferation, but when degenerative changes were more evident, smaller and more abundant vacuoles were seen. Also, the plasma membrane was altered and disrupted in some fibers, whereas many mitochondria showed a swollen appearance and, in some cells, they were disrupted. An amorphous extracellular material was also observed as a consequence of envenomation. It is proposed that the ultrastructural basis of the muscular necrosis induced by terciopelo venom is the disruption and severe alteration of the myofibril architecture.

マグロ類ジェリーミートの研究-II.

In order to see the extent of a destruction of muscle tissue occurring in the jellied meat, the so called “azuki”, of yellowfin tuna, the susceptibility of the jellied muscle to distintegration during homogenizing was examined by microscopic observation of the size and shape of the fragments released, and by a determination of myofibrillar protein extractability thereof. The jellied muscle was much more readily disintegrated into myofibrillar fragments than normal muscle. And, myofibrillar proteins in the jellied muscle were more easily extracted than those in normal muscle. These results indicate that the muscle tissue in jellied condition is very fragile. A microscopic observation of the fragments released from the jellied muscle showed that the architecture of myofibrils was loosened and/or broken in I band region, possibly at Z disk. Furthermore, a weakening of lateral attachment between myofibrils was suggested.