Scanning electron-microscopic studies on the three-dimensional structure of sarcoplasmic reticulum in the mammalian red,white and intermediate muscle fibers

Summary The three-dimensional structure of the sarcoplasmic reticulum (SR) in the red, white and intermediate striated muscle fibers of the extensor digitorum longus muscle of the rat was examined under a field-emission type scanning electron microscope after removal of cytoplasmic matrices by the osmium-DMSO-osmium procedure.In all three types of fibers, the terminal cisternae and transverse tubules form triads at the level of the A-I junction. Numerous slender sarcotubules, originating from the A-band side terminal cisternae, extend obliquely or longitudinally and form oval or irregular shaped networks of various sizes in front of the A-band, then become continuous with the tiny mesh (fenestrated collar) in front of the H-band. The A-and H-band SR appears as a single sheet of anastomotic tubules. Numerous sarcotubules, originating from the I-band side terminal cisternae, extend in threedimensional directions and form a multilayered network over the I-band and Z-line regions. At the I-band level, paired transversely oriented mitochondria partly embrace the myofibril. The I-band SR network is poorly developed in the narrow space between the paired mitochondria, but is well developed in places devoid of these mitochondria.The three-dimensional structure of the SR is basically the same in all three muscle fiber-types. However, the SR is sparse on the surface of mitochondria, so the mitochondria-rich red fiber has a much smaller total volume of SR than the mitochondria-poor white fiber. Moreover, the volume of SR of the intermediate fiber is intermediate between the two.     

High -resolution scanning electron-microscopic studies on the three-dimensional structure of mitochondria and sarcoplasmic reticulum in the different twitch muscle fibers of the frog

Summary The three-dimensional structure of the mitochondria and sarcoplasmic reticulum (SR) in the three types of twitch fibers, i.e., the red, white and intermediate skeletal muscle fibers, of the vastus lateralis muscle of the Japanese meadow frog (Rana nigromaculata nigromaculata Hallowell) was examined by high resolution scanning electron microscopy, after removal of the cytoplasmic matrices.The small red fibers have numerous mitochondrial columns of large diameter, while the large white fibers have a small number of mitochondrial columns of small diameter. In the medium-size intermediate fibers, the number and diameter of the mitochondrial columns are intermediate between those of the red and white fibers.In all three types of fibers, the terminal cisternae and transverse tubules form triads at the level of each Z-line. The thick terminal cisternae continue into much thinner flat intermediate cisternae, through a transitional part where a row of tiny indentations can be observed. Numerous slender longitudinal tubules originating from the intermediate cisternae, extend longitudinally or obliquely and form elongated oval networks of various sizes in front of the A-band, then fuse to form the H-band collar (fenestrated collar) around the myofibrils. On the surface of the H-band collar, small fenestrations as well as tiny hollows are seen. The three-dimensional structure of SR is basically the same in all three muscle fiber-types. However, the SR is sparse on the surface of mitochondria, so the mitochondria-rich red fiber has a smaller total volume of SR than the mitochondria-poor white fiber. The volume of SR of the intermediate fiber is intermediate between other the two.     

Scanning electron-microscopic study of lens fibers of the pig

Summary Whole pig lenses were fixed, critical-point dried and fractured, and the internal surfaces examined with a scanning electron microscope. At various locations from the equator to the center of the lens four types of fibers can be distinguished. The superficial fibers have small interdigitations. Cortical lens fibers, which are hexagonal in shape exhibit well developed ball-and-socket junctions. Other cortical fibers appear slightly undulated and show fine granulations. The core lens fibers are characterized by microplicae on the cell surfaces and by a more rounded or rectangular form. Results are discussed in relation to previous electron-microscopic studies of other species.     

The three-dimensional structure of motor endplates in different fiber types of rat intercostal muscle

Summary The three-dimensional organization of the motor end plates in the red, white and intermediate striated muscle fibers of the rat intercostal muscle was observed under a field-emission type scanning electron microscope after removal of connective tissue components by HCl hydrolysis.The motor endplate of the white fiber had terminal branches (or axon terminals), which were large, long and thin, and small but numerous nerve swellings (or terminal boutons). The motor endplate of the red fiber had terminal branches, which were small, short and thick, and had large but fewer nerve swellings. The motor endplate of the intermediate fiber was intermediate in size and structure between these two. In detached nerve-ending preparations, primary synaptic grooves with slit-like openings of the junctional folds appeared on the surface of the muscle fibers. The primary synaptic grooves were more developed in the white fiber than in the red fiber, and they were intermediate in the intermediate fiber. The numerical ratio of slit-like openings was 11.83.5 in the red, intermediate and white fiber, respectively.The Schwann cells and their processes were observed on the surface of the motor endplate, with the processes covering the upper orifices of the primary synaptic grooves and sealing the terminal branches. The number of Schwann cells was usually three in the white fiber, two in the intermediate fiber and one in the red fiber.     

High resolution scanning electron-microscopic study on the three-dimensional structure of the sarcoplasmic reticulum in the slow (tonic) muscle fibers of the frog,Rana nigromaculata

Summary The three-dimensional structure of the sarcoplasmic reticulum (SR) in the slow (tonic) fibers of the reclus abdominis muscle of the Japanese meadow frog (Rana nigromaculata nigromaculata Hallowell) was examined by high resolution scanning electron microscopy, after removal of the cytoplasmic matrices by the osmium-DMSO-osmium procedure. The SR forms a repetitive network throughout these fibers. At the level of the Z-line, a slender transverse tubule (T-tubule) runs transversely to the longitudinal axis of the myofibril. Small, spherical or ovoid terminal cisternae couple laterally with the T-tubule at intervals of 0.4–1.0 m, and form a terminal cisterna-T-tubule complex on whose surface tiny indentations are occasionally seen. Each terminal cisterna gives rise to a few sarcotubules that run in various directions, divide frequently and form circular or oval meshes of diverse sizes in front of the A- and I-bands. The sarcotubules usually form small meshes in the middle of the A-band, but occasionally fuse and form a poorly developed H-band (fenestrated) collar.     

Subsarcolemmal mitochondria and capillarization of soleus muscle fibers in young rats subjected to an endurance training

Summary Rats, 6 weeks old, were subjected to a program of endurance running for 3, 6 and 12 weeks. 0.5 to 0.8 m thick sections of Epon embedded soleus muscles were studied with morphometric methods.In cross-sections the area occupied by subsarcolemmal mitochondria was independent of the age, but was 53% higher after 12 weeks of training. The mean depth of the zones with subsarcolemmal mitochondria increased only 15% to about 0.9 m. Thus, the subsarcolemmal mitochondria showed a pronounced spreading at the muscle fiber surface in trained muscles. — The number of capillaries per fiber decreased slightly in controls and increased not significantly in trained muscles.It is concluded that the subsarcolemmal mitochondria supply the energy for the active transport of metabolites through the sarcolemma in oxidative muscle fibers, and that they are the limiting factor for endurance performance of the soleus muscle fibers because the changes in the capillarization were only small. It is suggested that the subsarcolemmal and the interfibrillar mitochondria have different functions and may therefore represent different types of mitochondria which can be distinguished by their morphology as well as by their biochemical properties.     

Scanning electron-microscopic study on the three-dimensional structure of motor endplates of the slow (tonic) muscle fibers in the frog,Rana n. nigromaculata

Summary The three-dimensional organization of the motor endplates of the slow fibers of the rectus abdominis muscle in the Japanese meadow frog (Rana nigromaculata nigromaculata Hallowell) is visualized by use of a field-emission scanning electron microscope after removal of connective tissue components by HCl hydrolysis. Clusters of shallow oval depressions 1–3 m in diameter are seen in the postsynaptic membrane at intervals of about 150 m. On the surface of these depressions, a few low bulges of postsynaptic membrane are irregularly arranged. Terminal boutons, 1–3 m in diameter, occur along the length of nerve branches and terminals and fit into the shallow oval depressions of the postsynaptic membrane. The Schwann cells covering the terminal branches exhibit a simpler organization than those in twitch fibers.     

Scanning electron-microscopic study of Reissner's fiber in the caudal spinal cord of the cat and rabbit

Summary The caudal portion of Reissner's fiber was examined by scanning electron microscopy (SEM) in the spinal cord of the cat and rabbit. In some preparations of both species the fiber displayed in the sinus terminalis of the central canal either stump-like terminations or structural modifications such as knot-like swellings and convolutions. In the same area homogeneous material could also be found, which obviously originated from the disintegrating fiber.     

Scanning electron microscopic studies on the synaptic bodies in the rat retina

Summary The ultrastructure of the synaptic bodies in the outer and inner plexiform layers of the rat retina was studied with scanning and transmission electron microscopy.The synaptic bodies in the outer plexiform layer are pear-shaped and their vitreal pole invaginated by processes from nerve cells. Their surfaces are covered with extracellular material, which is partly dissolved or redistributed during the fixation and rinsing procedure. The internal structure of the synaptic bodies is described.The synaptic bodies in the inner retinal plexiform layer are more difficult to identify with the scanning electron microscope. They are polyhedronal and also covered with extracellular material.The observations are discussed. The value of the application of two different preparation and analyzing methods, i. e. the scanning and the transmission electron microscopy, is stressed.Supported by grants from the Swedish Medical Research Council (B70-12X-2543-02), Expressens prenatalforskningsfond and Riksföreningen mot Cancer (265-B69-01X).     

Scanning electron-microscopic study of sperm retention and migration in the vagino-cervical region of the rabbit

Summary The pattern of sperm retention and migration in the vagino-cervical region of rabbit was studied by means of scanning electron microscopy. The following phenomena were observed: 1) Spermatozoa located in the vagina and at the orifice of the ectocervix are usually distributed diffusely. They appear to be resting on the epithelial surface; many are structurally abnormal or decapitated. 2) The great majority of spermatozoa, however, seems to be anchored or retained in narrow epithelial channels with their heads in close file formations. This phenomenon was observed particularly in the fornix vaginae as late as 24 h post coitum. 3) A great number of spermatozoa invading the cervix evidently migrates in groups along the mucosal surface. Their heads are oriented toward the uterus and contact the epithelial cells. Spermatozoa that migrate beyond the cervico-uterine junction are distributed in the same manner. 4) Spermatozoa colonizing the cervical crypts appear to be attached via the anterior margins of their heads to the epithelial cells or to the tips of kinocilia. Their tails project into the crypt lumen. It is suggested that mainly three factors may be responsible for these phenomena: (i) the fact that only motile spermatozoa overcome the vagino-cervical barrier; (ii) the tendency of spermatozoa to move along the mucosa in close vicinity to the epithelial cells; and (iii) the inability to recognize mechanical barriers on the migration route (e.g., cervical crypts) and to overcome them quickly. This may be one of many possible causes leading to sperm retention in the vagino-cervical region.Supported by a grant from the Deutsche Forschungsgemeinschaft     

The fine structure of red and white myotomal muscle fibres of the coalfish (Gadus virens)

Summary The fine structure of the red and white myotomal muscles of a marine teleost, the coalfish Gadus virens, has been examined and ultrastructural measurements and analyses carried out. The sarcomere lengths of the red and white fibres were found to be 1.60 minimum, 1.82 maximum and 1.70 minimum, 1.85 maximum, respectively. No significant difference was found between the red and white fibres in their percentage of sarcoplasmic reticulum and T system. Both were found to have regularly occurring triads at the Z disk level, to have distinctive M lines and to be multiply innervated. Ultrastructurally the two fibres can be distinguished by the thicker Z line and more abundant mitochondria of the red fibre, and by the ribbon-shaped peripheral myofibrils of the white fibres. The structure of the fibres in these two types of muscle is discussed in relation to their possible role in swimming.This work was supported by a research grant from the National Environmental Research Council.     

Stereological analyses of the structure of mitochondria in pigeon skeletal muscle

Summary The mitochondria in type-I and -II muscle fibres in the pectoralis major muscle of the pigeon (Columba livia) have been analysed using stereological techniques not previously applied in muscle biology.Mitochondrial volume fractions (V_V) were estimated in different regions of each type of muscle fibre using randomly orientated sampling sectors within fibre profiles. These sectors were sub-divided into smaller sampling regions to provide accurate data on the intracellular distribution of mitochondria. Estimates of the external surface densities of mitochondria per unit volume of fibre, S_{V total} ^surface , and also the densities of mitochondrial cristae, S_{V total} ^cristae , were obtained using a specific technique derived for analysing anisotropic structures (Saltykov, 1958). The relative amounts of the random and orientated mitochondrial membranes were also estimated.Significiant differences were found to exist between the different types of muscle fibres and considerable though constant variations in the intracellular arrangement of mitochondria were also found.     

Scanning electron microscopic study on the structure of the lingual papillae of the Saanen goat

The morphology of lingual papillae of the ten male mature Saanen goats (11 months old, approximately 42 kg in weight and of a known pedigree) was examined by scanning electron microscopy. Tissues were taken from the dorsal and ventral surfaces of the apex, body and root of the tongue, and were prepared accordingly and observed under the scanning electron microscope. On the dorsal and ventro-lateral surfaces of the lingual mucosa, three types of mechanical papillae (filiform, lenticular, and conical) and two types of gustatory papillae (vallate and fungiform) were observed. The structure and density of the filiform papillae differentiated on the anterior, posterior and ventro-lateral aspects of the tongue. Two types of lenticular papillae, both possessing a prominent surrounding papillary groove, were determined. The pyramidal-shaped type I lenticular papilla had a pointed apex while the round-shaped type II lenticular papilla possessed a blunt apex. Certain number of the type I lenticular papillae had double apices. The larger conical papillae were hollow structures, differing structurally from the filiform papillae with their larger size, a tip without projections and lack of the secondary papillae. The vallate papillae were present on both rims of the torus linguae, were encircled by a prominent gustatory furrow which was also surrounded by a thick annular fold. The fungiform papillae were scattered among the filiform papillae in the anterior two-thirds of the dorsal and lateral surfaces, and each of them was highly protected by surrounding filiform papillae, yet encircled by a papillary groove. Our findings indicate that Saanen goat have profuse distribution of papillae on the tongue displaying morphological features characteristic of mechanical function.     

Effect of cold environment on skeletal muscle mitochondria in growing rats

Summary Growing rats (4 weeks old) were kept for 3 weeks at 11° C and 24° C respectively. The cold-adapted animals showed a significantly higher oxygen consumption (64%). Volume density of subsarcolemmal and interfibrillar mitochondria as well as volume density of fat droplets were estimated in M. soleus and the diaphragm of both groups. In cold-adapted animals, the total volume of mitochondria was significantly increased by 24% in diaphragm and 37% in M. soleus. The volume of subsarcolemmal mitochondria was almost doubled in each muscle, but the volume of interfibrillar mitochondria did not change significantly. The surface of the inner mitochondrial membranes per unit volume of mitochondrion in M. soleus was significantly increased both in interfibrillar and subsarcolemmal mitochondria, whereas the surface of the outer mitochondrial membranes per unit volume of mitochondrion was increased only in the subsarcolemmal mitochondria. The volume of fat droplets in the diaphragm and M. soleus of cold adapted animals increased significantly by 62% and 150% respectively.     

Scanning electron microscopy of the muscle coat of the guinea-pig small intestine

Summary We have studied the layers of the muscular coat of the guinea-pig small intestine after enzymatic and chemical removal of extracellular connective tissue. The cells of the longitudinal muscle layer are wider, have rougher surfaces, more finger-like processes and more complex terminations, but fewer intercellular junctions than cells in the circular muscle layer. A special layer of wide, flat cells with a dense innervation exists at the inner margin of the circular muscle layer, facing the submucosa. The ganglia of the myenteric and submucosal plexuses are covered by a smooth basal lamina, a delicate feltwork of collagen fibrils, and innumerable connective tissue cells. The neuronal and glial cell processes at the surface of ganglia form an interlocking mosaic, which is loosely packed in newborn and young animals, but becomes tightly packed in adults. The arrangement of glial cells becomes progressively looser along finer nerve bundles. Single varicose nerve fibres are rarely exposed, but multiaxonal bundles are common. Fibroblast-like cells of characteristic shape and orientation are found in the serosa; around nerve ganglia; in the intermuscular connective tissue layer and in the circular muscle, where they bridge nerve bundles and muscle cells; at the submucosal face of the special, flattened inner circular muscle layer; and in the submucosa. Some of these fibroblast like cells correspond to interstitial cells of Cajal. Other structures readily visualized by scanning electron microscopy are blood and lymphatic vessels and their periendothelial cells. The relationship of cellular elements to connective tissue was studied with three different preparative procedures: (1) freeze-cracked specimens of intact, undigested intestine; (2) stretch preparations of longitudinal muscle with adhering myenteric plexus; (3) sheets of submucosal collagen bundles from which all cellular elements had been removed by prolonged detergent extraction.     

Capillarization,mitochondrial densities,oxygen diffusion distances and innervation of red and white muscle of the lizard Dipsosaurus dorsalis

Summary The white and red regions of the iliofibularis muscle of the lizard Dipsosaurus dorsalis were analyzed using histologic and morphometric analysis. These regions are composed of fast glycolytic (FG) and both fast oxidative, glycolytic (FOG) and tonic fibers, respectively. Endplate morphology and number of endplates per fiber were estimated from fibers from both areas. Capillary volume densities of the red and white regions were quantified from transverse sections. Mitochondrial volume of fibers from the red and white regions were estimated from electron micrographs.All fibers from the white region of the iliofibularis possessed a single, well defined endplate, as did most red region fibers. The remaining red fibers (28±5%) possessed an average of 14.7±3 endplates each, distributed along the entire length of the fiber at intervals of approximately 1124 m.Red fibers possessed twice the mitochondrial volume of white fibers (7.6±0.4%, red; 3.8±0.3%, white). Mitochondria were distributed uniformly through the fibers from both regions. Capillary anisotropy was low ( = 1.018) in both regions. Capillary densities of the red region (629±35 mm^-2) were much greater than those of the corresponding White region (73±8 mm^-2).The data indicate that capillary densities, mitochondrial volumes and theoretical diffusion distances correlate well with the oxidative capacity of lizard muscle fibers. Tonic fibrs of this species appear oxidative and therefore metabolically capable of functioning during locomotion. The similar mitochondrial volumes and capillary densities of reptilian and mammalian muscles suggest that the greater oxidative capacity of mammalian muscle is due in part to possession of more oxidatively active mitochondria rather than to possession of more mitochondria per se.     

Architecture of the media of the arterial vessels in the dog brain: A scanning electron-microscopic study

Summary The architecture of the media of arterial vessels in dog brain was investigated using scanning electron microscopy. The arrangement and shape of the circularly-oriented smooth muscle cells varied with vessel diameter: The arteries (>100 m in diameter) had 4–10 layers of spindle-shaped smooth muscle cells; the muscular arterioles (30–100 m), 2–3 layers of spindle-shaped smooth muscle cells; the terminal arterioles (10–30 m), a compact layer of spindle-shaped smooth muscle cells with more dominant nodular or rod-like processes and thin lateral processes; and the precapillary arterioles (5–15 m), a less compact layer of branched smooth muscle cells.Longitudinally-oriented muscles were observed in the medio-adventitial border. The distribution and arrangement of these muscles varied with vessel size: in the large arteries (> 300 m in diameter), at the branching sites only; in the small arteries (100–300 m), at both the branching and non-branching sites; in the muscular arterioles, at both the branching and non-branching sites in a reticular arrangement with some muscle cells having an asteroid appearance; in the terminal aterioles, only asteroid-like muscle cells were found at the branching and non-branching sites.     

The effect of temperature on conduction velocity in human muscle fibers

The effects of variation of intramuscular temperature (T) on conduction velocity (CV) of the action potential along single human muscle fibers of the biceps brachii was studied in situ in 15 normal volunteers (mean age 39 years, range 21–62 years). Cooling was obtained by direct application of ice over a rectangular skin region including the stimulating and recording area. The intramuscular T was monitored by a needle thermocouple (copperconstantane). In all the 24 muscle fibers studied, a linear relationship was observed between CV and T. The slopes of the regression lines, ranging between 0.190 and 0.079 m/s, were positively correlated with the starting CV at 36°C ranging between 2.2 and 5.2 m/s. If conduction changes are expressed as a percentage of the basal CV at 36°C, the CV/T coefficient is the same for all the fibers and independent of the individual CV: 3.4% of CV/°C.     

Fine structure and metabolic adaptation of red and white muscles in tuna

Synopsis An electron microscopic study of the red and white muscle fibres in the trunk musculature of the Kawakawa tuna (Euthynnus affinis) was carried out with a view to correlating their structure with metabolic adaptation. The red fibres which are considerably smaller in diameter (34.58 m ± 6.16 S.D.) are characterized by their high content of myoglobin, mitochondria, lipid droplets and glycogen granules. The white fibres which are relatively larger in diameter (66.03 m ± 11.59 S.D.) are characterized by their lack of myoglobin, low mitochondria) density, high content of glycogen granules and the conspicuous absence of lipid droplets. The characteristics in fine structure of the two fibre types are discussed in the light of their metabolic adaptation, the red fibres as being adapted for long term cruising movement utilizing lipid as the main source of energy and the white fibres for short bursts of activity metabolizing glycogen as the chief fuel.The tuna, with the acquisition of the counter-current heat exchange system which provides for the retention of the heat generated from high substrate oxidation in the red muscle and an efficient respiratory system, it is postulated, is well adapted for high speed sustained swimming.