Strong Succinate Dehydrogenase Research Articles

The claw closer muscle of two estuarine crab species,Cyrtograpsus angulatusandNeohelice granulata(Grapsoidea, Varunidae): histochemical fibre type composition

Longo, M.V. and Díaz, A.O. (2011). The claw closer muscle of two estuarine crab species, Cyrtograpsus angulatus and Neohelice granulata (Grapsoidea, Varunidae): histochemical fibre type composition. —Acta Zoologica (Stockholm) 00: 1–7. This study permitted the characterization of four types of muscle fibres in the claw closer muscles of Cyrtograpsus angulatus and Neohelice granulata. Succinic dehydrogenase (SDH) for mitochondria, periodic acid Schiff (PAS) for glycogen, Sudan Black B for lipids and myosin-adenosine triphosphatase (m-ATPase) preincubated at alkaline and acid pHs were used for that purpose. The mean fibre diameters, the relative areas and frequencies of each muscle fibre type were calculated. Types I and IV would be considered ‘extreme’ groups with type I fibres large, weak and acid/alkaline-labile m-ATPase, weak SDH, PAS and Sudan, and type IV fibres small, very strong and acid/alkaline-resistant m-ATPase, strong SDH and PAS, and moderate Sudan. Types II and III would belong to a predominant ‘intermediate’ group. Type IV fibres were scarce in C. angulatus but represented 25% of the total fibre population in N. granulata. In C. angulatus, the relative area occupied by type I fibres was bigger than its relative proportion, whereas in N. granulata, types I and II had similar patterns. Concluding, variations in fibre type composition in the claw closer muscles of C. angulatus and N. granulata would be linked to different habitats and feeding behaviours.

A Novel Heteroplasmic tRNASer(UCN) mtDNA Point Mutation Associated With Progressive Ophthalmoplegia and Dysphagia

We report a novel heteroplasmic mitochondrial DNA mutation in the tRNA gene at nucleotide 7458 (m.7458G>A) in a 26-year-old patient affected with sporadic progressive external ophthalmoplegia associated with dysphagia. Muscle biopsy showed a strong succinate dehydrogenase staining, ragged red fibers, and 15% of cytochrome c oxidase-negative fibers. Activities of mitochondrial respiratory chain complexes I+III and IV were reduced. The mutation was heteroplasmic (75%) in the muscle, but undetectable in accessible tissues from the patient and his maternal relatives. This report expands the molecular heterogeneity of progressive external ophthalmoplegia.

Delayed mitochondrial dysfunction in apoptotic hair cells in chinchilla cochleae following exposure to impulse noise

Apoptotic death of hair cells (HCs) in the cochlea has been found following exposure to intense noise. The current study was designed to examine the mitochondrial energetic function of HCs during the course of noise-induced apoptosis. Two aspects of the mitochondrial energetic function, succinate dehydrogenase (SDH) activity and mitochondrial membrane potential (MMP), were examined in HCs of chinchilla cochleae following exposure to a series of 75 pairs of impulse noises at 155 dB pSPL. The results showed that nuclear condensation and uptake of propidium iodide or trypan blue appeared at 10 min after the noise exposure, indicating a rapid progression of HC apoptosis. However, SDH activity was preserved at this time point. As the time elapsed (1 hr or 24 hrs) after the noise exposure, all newly-generated apoptotic HCs showed strong SDH activity, indicating the preservation of SDH activity during the course of apoptosis. Examination of MMP with rhodamine 123 staining revealed that MMP was sustained in the apoptotic HCs having mild nuclear condensation, even after the occurrence of cell membrane leakage. MMP was reduced with further progression of nuclear condensation. These results suggest the presence of a delayed mitochondrial dysfunction in apoptotic HCs following exposure to intense noise.

Pathology of Mitochondrial Encephalomyopathies

Muscle biopsy provides the best tissue to confirm a mitochondrial cytopathy. Histochemical features often correlate with specific syndromes and facilitate the selection of biochemical and genetic studies. Ragged-red fibres nearly always indicate a combination defect of respiratory complexes I and IV. Increased punctate lipid within myofibers is a regular feature of Kearns-Sayre and PEO, but not of MELAS and MERRF. Total deficiency of succinate dehydrogenase indicates a severe defect in Complex II; total absence of cytochrome-c-oxidase activity in all myofibres correlates with a severe deficiency of Complex IV or of coenzyme-Q10. The selective loss of cytochrome-c-oxidase activity in scattered myofibers, particularly if accompanied by strong succinate dehydrogenase staining in these same fibres, is good evidence of mitochondrial cytopathy and often of a significant mtDNA mutation, though not specific for Complex IV disorders. Glycogen may be excessive in ragged-red zones. Ultrastructure provides morphological evidence of mitochondrial cytopathy, in axons and endothelial cells as well as myocytes. Abnormal axonal mitochondria may contribute to neurogenic atrophy of muscle, a secondary chronic feature. Quantitative determinations of respiratory chain enzyme complexes, with citrate synthase as an internal control, confirm the histochemical impressions or may be the only evidence of mitochondrial disease. Biological and technical artifacts may yield falsely low enzymatic activities. Genetic studies screen common point mutations in mtDNA. The brain exhibits characteristic histopathological alterations in mitochondrial diseases. Skin biopsy is useful for mitochondrial ultrastructure in smooth erector pili muscles and axons; skin fibroblasts may be grown in culture. Mitochondrial alterations occur in many nonmitochondrial diseases and also may be induced by drugs and toxins.

Insulin edema in diabetes mellitus associated with the 3243 mitochondrial tRNA Leu(UUR) mutation; Case reports

We encountered a patient with diabetes mellitus due to the 3243 mitochondrial tRNA mutation(DM-Mt3243), who developed insulin edema and hepatic dysfunction after starting insulin. Such a rare phenomenon was unlikely to be a fortuitous coincidence in mitochondrial diabetes, as none in 197 non-mutant NIDDM patients had same episode. Moreover, similar leg edema was noticed in another DM-Mt3243 patient, and other two DM-Mt3243 patients had leg edema which responded to coenzyme Q10. These observations suggest further a role of mitochondrial function on leg edema. The mechanism of his insulin edema may involve vasomotor changes induced by the rapidly glycemic control, because our case of insulin edema had a prominent increase of strong succinate dehydrogenase reactive vessels. Alternatively, myocardial dysfunction might have produced leg edema and hepatic dysfunction, because he had subclinical myocardial dysfunction, judged by imaging with β-methyl- p-( 123I)- iodophenyl-pentadecanoic acid. The third explanation is that a rapid improvement of glycemic control might have induced hepatic reoxygenation and the production of reactive oxygen species in the liver that contributed to cell damage. Thus, although we cannot draw definite conclusion, our experiences here suggest that mitochondrial dysfunction is important in the etiology of insulin edema.

Macrophages of the rat thymus after cyclosporin treatment

Young adult Wistar rats received 40 mg/kg of cyclosporin perorally for 21 days. Cyclosporin induced almost total disappearance of thymic medulla, whereas the cortex remained preserved. Although the density of cortical macrophages did not change significantly, their characteristics altered markedly and they became enlarged and rounded. In addition to an increase in acid phosphatase and nonspecific esterase activities, cortical macrophages developed very strong succinic dehydrogenase and chloroacetate esterase activities and a fine, granular, aldehyde fuchsin-positive cytoplasmic content. However, these cytoplasmic granules were PAS-negative and were not sudanophilic. Cortical macrophages retained their normal antigenic properties (which were studied by the use of ED1, ED2 and R-MC 41 monoclonal antibodies). Phagocytic cells in the remaining medullary islands retained their usual characteristics. The changes in cortical macrophages after cyclosporin treatment are discussed, especially in relation to the characteristics of macrophages of the cortico-medullary zone in the normal rat thymus.

Fibre-optic spectrophotometry of immature bovine skeletal muscles and the cellular distribution of myoglobin and succinate dehydrogenase.

Samples of diaphragm and pectoralis profundus were taken from nine calves with a range of blood haemoglobin levels of 4 to 8.5 g/100 ml. In both muscles, fibres with strong succinate dehydrogenase activity contained myoglobin, but in the pectoralis there were many fibres with strong alkaline ATPase activity and weak succinate dehydrogenase activity that had low or undetected levels of myoglobin. The whole cross-sectional area of individual fibres was scanned to map the distribution of succinate dehydrogenase activity. Among fibres with similar levels of ATPase activity, those from the diaphragm had greater succinate dehydrogenase activity than those from the pectoralis. Subsarcolemmal succinate dehydrogenase activity was greater than the axial succinate dehydrogenase activity, and radial gradients of succinate dehydrogenase activity were steepest in the diaphragm. For pectoralis fibres with weak ATPase, the mean and the axial succinate dehydrogenase activities were correlated with blood haemoglobin levels (r = 0.62 and r = 0.61, respectively; P less than 0.05 with a Student's t-test). Muscle colour was measured directly by fibre-optic spectrophotometry and correlations of absorbance with succinate dehydrogenase activity were obtained. Absorbance at 620 nm 24 h post-mortem was correlated with succinate dehydrogenase activity in pectoralis fibres with weak ATPase (r = 0.81; P less than 0.005).

Enzyme-histochemical characterization of macrophages in the rat thymus, with special reference to metallophilic cells of the corticomedullary zone.

The cortex and medulla of the normal rat thymus are populated with scattered cells which are strongly positive for acid phosphatase and only weakly (cortex) or moderately (medulla) positive for nonspecific esterase. Corticomedullary zone is characterized, as a specific entity within the thymic tissue, by the presence of very large macrophages, highly positive for acid phosphatase and nonspecific esterase. Moreover, these cells, located exclusively in the corticomedullary zone, show very strong succinic dehydrogenase activity and contain the intracytoplasmic granules of varying size, which are aldehyde fuchsin, PAS, and oil red O positive. The presence of cholesterol was demonstrated within the lipid content.

The radial distribution of succinate dehydrogenase activity in porcine muscle fibres

A microscope photometer with a computer-controlled scanning stage was used to map the distribution of succinate dehydrogenase (SDH) activity in transverse sections of skeletal muscle fibres of pigs from 9 to 29 weeks of age. Absorbance due to SDH activity was measured in successive concentric zones that converged on the central axis of the muscle fibre. In all fibres, there was less SDH activity in the axis of the fibre than in the periphery of the fibre. In fibres with strong adenosine triphosphatase (ATPase) activity and weak overall SDH activity, the radial gradient of SDH activity remained constant as fibres grew in cross-sectional area. However, in fibres with weak ATPase and strong SDH and in fibres with strong ATPase and strong SDH, the radial gradient of SDH activity increased as fibres grew larger. Changes in radial gradients were due to both decreased SDH activity in the axis and to increased SDH activity in the periphery. In all three fibre types, the overall SDH activity per fibre increased with age.

Intracellular distribution of succinate dehydrogenase activity in skeletal muscle fibers of geese

Samples of iliotibialis anterior and pectoralis muscles were taken from five ganders (Anser domesticus). Serial transverse sections were reacted for succinate dehydrogenase (SDH) and alkali-stable adenosine triphosphatase (ATPase). The distribution of SDH activity within individual muscle fibers was measured with a scanning photometer. In many individual fibers, SDH activity was stronger in the periphery than in the axis. This gradient was steepest (−0.034 ± 0.019 absorbance units per concentric zone of 2 μm diameter measurements) in pectoralis fibers with strong SDH activity. In the pectoralis, radial gradients were correlated with fiber area so that the smallest fibers tended to have the steepest gradients of SDH activity. However, this relationship was reversed in fibers with strong ATPase and weak SDH activity in the iliotibialis anterior, and the largest fibers tended to have the steepest gradients. In all fiber types of both muscles, fibers with greater mean SDH activity tended to have steeper gradients.

The histochemistry of very small muscle fibres in growing skeletal muscles

Serial sections of biceps femoris muscles from 10 rapidly growing pigs were reacted for succinate dehydrogenase (SDH) and myofibrillar adenosine triphosphatase (ATPase) and were stained with silver to delineate the endomysial boundaries of their muscle fibres. The histochemistry of very small fibres (<0.001 mm 2) was similar to that of surrounding fibres with a normal diameter. Of the small fibres, 71.5% had strong ATPase, 27.5% had weak ATPase, 22% had strong SDH, 23.8% had intermediate SDH and 54.1% had weak SDH reactions. Corresponding values for surrounding fibres with a normal diameter were 87.9% with strong ATPase, 11.8% with weak ATPase, 35.1% with strong SDH, 14.5% with intermediate SDH, and 50.5% with weak SDH reactions. An appreciable number of small fibres were histochemically unrelated to any of their surrounding fibres: 11.0% for ATPase, 12.8% for SDH, and 5.5% for both ATPase and SDH. The cross-sectional shapes of small fibres were similar to those of their surrounding fibres. It was concluded that these small fibres were probably the tapered ends of intrafascicularly terminating muscle fibres rather than new muscle fibres formed by splitting.

Histochemical and immunohistochemical studies of the cerebellum from the reeler mutant mouse.

The organization of the fiber connections and architecture of the cerebellum of reeler mutant mice was analyzed by an immunohistochemical and histochemical procedure. By immunohistochemical staining of the myelinated fiber arborization with antiserum against myelin basic protein, it was found in the reeler cerebellum that the fibers ran in all directions throughout the white matter. Some of the fibers surrounded Purkinje cells. The distribution of AChE-positive fibers was abnormal and some Purkinje cells surrounded by AChE-positive fibers were present. Molecular layers and glomeruli showed strong succinic dehydrogenase (SDH) activity. The white matter of the reeler cerebellum showed a mosaic of SDH-positive and -negative sites. The results indicate that changes in the distribution of myelinated fiber arborization and the change in the distribution of SDH and AChE activity are different in each part of the cerebellum as a result of the disorganization of the architecture of the reeler mutation.

Low temperature activation ofpost mortem glycogenolysis in bovine skeletal muscle fibres

Trimmed strips of sternomandibularis muscles taken from freshly-slaughtered cattle were placed in an isotonic myograph and cooled to 1 degree C. Spontaneous activity due to neuromuscular irritability was minimized by keeping muscle surfaces moist and anaerobic and was monitored by electromyography. Muscle strips were removed and frozen for histochemical analysis after they had completed their initial phase of cold-induced shortening (several hours). Control strips maintained for an equal time at 24 degrees C rarely depleted the stainable glycogen in any of their muscle fibres so as to become PAS-negative. In chilled muscle strips, however, glycogenolysis was activated in some muscle fibres and they became PAS-negative. In serial sections, most of the PAS-negative fibres exhibited strong ATPase and weak succinate dehydrogenase activity. Fibres with weak ATPase and strong succinate dehydrogenase activity rarely became PAS-negative. These results are in agreement with biochemical reports of a cold-induced (less than 5 degrees C) activation of glycolysis in skeletal muscle post mortem. Investigations on untrimmed lengths of excised sternomandibularis muscle indicated that longitudinal muscle damage caused in cutting muscle strips for the myograph and/or their more rapid rate of initial cooling had facilitated the depletion of stainable glycogen.

A method for myoglobin in cryostat sections of muscle by precipitation with sulfosalicylic acid.

Transverse cryostat sections of skeletal muscle were fixed in a solution containing 1.5% glutaraldehyde and 1.5% sulfosalicylic acid and stained in a solution containing equal volumes of 3% hydrogen peroxide and 50% ethanol saturated with o-tolidine. Myoglobin in the sarcoplasm of muscle fibers was precipitated and stained blue. Applicability of this method to cryostat sections, without glutaraldehyde fixations prior to freezing, allowed the myoglobin content of individual muscle fibers to be correlated with other histochemical characteristics of the same fibers seen in serial sections. In the dark red bovine sternomandibularis muscle, fibers with weak adenosine triphosphatase (ATPase) and strong succinate dehydrogenase (SDH) activity always exhibited strong myoglobin staining. An equal degree of staining was found in many fibers with strong ATPase and intermediate to strong SDH activity. Fibers with strong ATPase and weak SDH activity were less strongly stained than the preceding types.

Changes in the mitochondria in the initial part of the axon during regeneration.

The initial part of the axon including the axon hellock, the initial unmyelimated segment and the beginning of myelinated axon was studied electron microscopically during regeneration, 1--30 days following a crush lesion of the rat hypoglossal nerve. Large mitochondria reaching 1.1 mum in diameter, with abundant cristae and dense granules in the matrix were observed between days 3--21. They formed clusters in the initial myelinated segment of the axon. End-to-end contacts and ribosomes around them were very often visible. The large mitochondria exhibited strong succinate dehydrogenase and NAD - H2 diaphorase activities. The relationship between the appearance of large and active mitochondria in the initial part of the axon and the elevated axonal transport during regeneration of the peripheral nerve is also discussed.

Histochemistry and the structure of the skin of a murrel, Channa striata (Bloch, 1797) (Channiformes, Channidae). I. Epidermis.

The functional organization and cytochemistry of the epidermis of an air-breathing fish, Channa striata, is described. In the basal cells a dense population of mitochondria, strong alkaline phosphatase, and succinic dehydrogenase activity indicates high metabolic activity. In the outermost layer, polygonal cells showed strong succinic dehydrogenase activity and elaborated sulfated acid mucopolysaccharides and lipids. Mucous cells were also numerous. A new term, 'sacciform granulated cells,' is proposed for the 'sacciform cells' or 'granular cells.' The contents of these cells are basic proteins. A thick coat of slime containing mucopolysaccharides, lipids, and basic proteins is probably important in keeping the skin moist for cutaneous respiration, retarding the rate of water loss by evaporation, facilitating burrowing in the mud and swimming movement in water, and protecting the skin from bacterial and fungal attacks. A relatively thin epidermis (32 μm on the general surface, 18 μm at the posterior free margins, and 6 μm below the scales) in conjunction with vascularization of the stratum laxum probably assists cutaneous respiration.

Histochemical observations on bone and periodontal tissues in the rat administered with cortisone and parathyroid hormones.

Histochemical study in epiphyseal cartilages and periodontal tissues of the rat subjected to cortisone and PTH administration was examined enzyme histochemically in the decalcified frozen sections with neutral EDTA. In cortisone and PTH-treated rats, altered proliferating cartilage cells in the epiphysis revealed a weak, and decreasing activities of oxidative enzymes with the comparison to those of the control, however, they showed similar activity for alkaline and acid phosphatases as normal cells. Epiphysis and alveolar bones in large dose of cortisone administered rats exhibited an absence or markedly decreasing number of osteoclasts, but on the contrary, those of PTH treated rats led to an increased number of the cells. Osteoclasts in hormone treated conditions were characterized by the existence of the strong succinate dehydrogenase activity as well as in normal status.

Histochemical mapping of succinic dehydrogenase and cytochrome oxidase in the diencephalon and basal telencephalic centers of the brain of squirrel monkey (Saimiri sciureus).

The distribution of succinic dehydrogenase (SDA) and cytochrome oxidase (Cy. O) was mapped in the various diencephalic nuclei and basal telencephalic centers of the squirrel monkey brain. Thirty μ thick formaldehyde fixed serial sections were also studied for the delineation of the various nuclei, but histochemical preparations proved equally useful for this purpose. Strong SDA and Cy. O activity were observed in the habenular, pulvinaris, anterior dorsalis and ventralis, lateralis dorsalis and posterior, and ventral posterior nuclei. The corpus geniculatum laterale and mediale also showed a strong reaction for these enzymes. The nucleus ventralis anterior, which occupies a very large area in the rostral part of the thalamus, showed moderately strong activity in the cellular patches and negligible activity in the thick fiber bundles passing through it. A comparatively weak reaction was observed in the midline thalamic nuclei. The nucleus caudatus and putamen, however, showed very strong SDA/Cy. O activity. The hypothalamic nuclei showed mild Cy. O and moderate SDA reactions, except for the nucleus ventromedialis hypothalami. The latter showed a little stronger enzyme reaction. The fibers of the internal capsule and the anterior commissure showed little SDA and mild Cy. O activity. The various nuclei of the amygdaloid complex showed similar histochemical reactions with moderate SDA and mild Cy. O activity.

Histochemical mapping of succinic dehydrogenase and cytochrome oxidase in the pons and mesencephalon of squirrel monkey (Saimiri sciureus).

The distribution of succinic dehydrogenase (SDA) and cytochrome oxidase (Cy. O) has been investigated in a series of sections through the pons and mesencephalon of the squirrel monkey brain. The localization of the two enzymes is very similar in the various regions and shows only slight differences. The epiphysis, however, shows moderately strong SDA and very mild Cy. O activity. Particularly strong SDA and Cy. O activity has been observed in the cell bodies of the various cranial nerve nuclei, nucleus colliculi inferioris, colliculi superioris, nuclei griseum pontis, reticularis tegmenti pontis, lemnisci lateralis pars dorsalis, geniculatum laterale and mediale, and pulvinaris. The enzyme content of the neurons and cell bodies is generally stronger compared to the neuropil which often occurs in smooth, loose, compact and reticulated forms. Any special relationship between the neurons and neuropil with regard to their enzyme content has, however, not been observed. The cranial nerves, and fibers of the brachium conjunctivum, corpus callosum, and fornix show very mild enzyme activity except those of the trapezoid complex which show moderate enzyme activity.

Histochemical mapping of succinic dehydrogenase and cytochrome oxidase in the spinal cord, medulla oblongata and cerebellum of squirrel monkey (Saimiri sciureus).

The distribution of succinic dehydrogenase (SDA) and cytochrome oxidase (Cy. O) in serial sections of the cervical region of the spinal cord and the medulla oblongata, arranged caudo-cranially, has been described. The motor cranial nerve nuclei exhibit strong SDA and Cy. O activity in the neurons and neuropil. The nuclei gracilis, cuneatus, olivaris inferior, cochlearis and vestibularis likewise show strong enzyme activity. Nucleus intercalatus and nucleus tractus solitarius, however, show weak and moderate enzyme activity respectively. The lateral part of formatio reticularis myelencephali shows less SDA and Cy. O compared to the medial part, which shows some accumulation of these enzymes in the neuropil. The neuropil of the molecular layer of cerebellar cortex and the perikarya and dendrites of the Purkinje cells show strong SDA and Cy. O activity. The granular layer exhibits stronger SDA and Cy. O in the synaptic glomeruli. The cerebellar nuclei possess stronger enzyme activity in the neurons and dendritic branches, compared to mild activity in the neuropil.