scholarly journals Needle muscle biopsy: technique validation and histological and histochemical methods for evaluating canine skeletal muscles

2017 ◽  
Vol 38 (2) ◽  
pp. 765
Author(s):  
Sérgio De Almeida Braga ◽  
Felipe Gomes Ferreira Padilha ◽  
Ana Maria Reis Ferreira
Keyword(s):  
Muscle Biopsy ◽  
Skeletal Muscles ◽  
Muscle Fibers ◽  
Gluteus Medius ◽  
Type I ◽  
Fiber Types ◽  
German Shepherd ◽  
Muscle Sample ◽  
Biopsy Technique ◽  
Histochemical Methods

This study evaluated the needle muscle biopsy technique using a 6G Bergström percutaneous needle combined with histological and histochemical methods to analyze the skeletal muscle of dogs. There are few studies about canine skeletal muscles and a lack of reports in the literature about tissue collection and analysis for canine species. Evaluation of 32 German Shepherd samples collected from the gluteus medius, at a depth of 3 cm, was performed. The choice of gluteus medius and the 3-cm depth provided good quantity fragments with sufficient sizes (3–5 mm), which permitted optimal visualization of muscle fibers. Myosin ATPase, at pH 9.4, 4.6, and 4.3, and SDH reactions revealed that all muscle samples analyzed had fibers in the classic mosaic arrangement, enabling counting and typification. The mean percentages of fibers were 29.95% for type I and 70.05% for type II. On the basis of these results, we concluded that the percutaneous needle biopsy technique for canine skeletal muscles is a safe and easy procedure that obtains fragments of proper sizes, thereby enabling the study of muscle fibers. Standardization of the muscle of choice and the depth of muscle sample collection significantly contributed to this success. This is an important method to evaluate muscle fiber types of dogs and diagnose important diseases affecting the skeletal muscles.

Beneficial effects of voluntary wheel running on the properties of dystrophic mouse muscle

1996 ◽  
Vol 80 (2) ◽  
pp. 670-679 ◽  
Author(s):  
A. Hayes ◽  
D. A. Williams
Keyword(s):  
Skeletal Muscles ◽  
Wheel Running ◽  
Voluntary Exercise ◽  
Mdx Mice ◽  
Free Access ◽  
Type I ◽  
Fiber Types ◽  
Force Output ◽  
Mouse Muscle ◽  
Beneficial Effects

Effects of voluntary exercise on the isometric contractile, fatigue, and histochemical properties of hindlimb dystrophic (mdx and 129ReJ dy/dy) skeletal muscles were investigated. Mice were allowed free access to a voluntary running wheel at 4 wk of age for a duration of 16 (mdx) or 5 (dy/dy) wk. Running performance of mdx mice (approximately 4 km/day at 1.6 km/h) was inferior to normal mice (approximately 6.5 km/day at 2.1 km/h). However, exercise improved the force output (approximately 15%) and the fatigue resistance of both C57BL/10 and mdx soleus muscles. These changes coincided with increased proportions of smaller type I fibers and decreased proportions of larger type IIa fibers in the mdx soleus. The extensor digitorum longus of mdx, but not of normal, mice also exhibited improved resistance to fatigue and conversion towards oxidative fiber types. The dy/dy animals were capable of exercising, yet ran significantly less than normal animals (approximately 0.5 km/day). Despite this, running increased the force output of the plantaris muscle (approximately 50%). Taken together, the results showed that exercise can have beneficial effects on dystrophic skeletal muscles.

Cross-reinnervated motor units in cat muscle. I. Flexor digitorum longus muscle units reinnervated by soleus motoneurons

1985 ◽  
Vol 54 (4) ◽  
pp. 818-836 ◽  
Author(s):  
R. P. Dum ◽  
M. J. O'Donovan ◽  
J. Toop ◽  
R. E. Burke
Keyword(s):  
Muscle Fibers ◽  
Motor Units ◽  
Muscle Fiber Types ◽  
Type I ◽  
Fiber Types ◽  
Glycogen Depletion ◽  
Flexor Digitorum Longus ◽  
Wet Weight ◽  
Flexor Digitorum ◽  
Individual Motor

The properties of flexor digitorum longus (FDL) muscles and of individual motor units were studied in cats 30-50 wk after self-reinnervation by FDL motoneurons (FDL----FDL) or cross-reinnervation by soleus (SOL) motoneurons (SOL----FDL). Individual motor units were functionally isolated by intracellular recording and stimulation of identified SOL alpha-motoneurons. Glycogen-depletion methods permitted histochemical study of muscle fibers belonging to physiologically characterized muscle units. The observations were compared with data from normal cat FDL muscles and motor units (27). Intentionally self-reinnervated FDL muscles (FDL----FDL; n = 5) were normal in size and wet weight. FDL----FDL motor units could be classified into the same physiological categories found in normal FDL [types: fast contracting, fatigable (FF), fast contracting, fatigue resistant (FR), and slow (S); n = 24], with approximately the same proportions as normal. The histochemical muscle fiber types associated with these categories were also qualitatively normal although there was evidence of marked distortion of the normal histochemical mosaic. These data confirm other studies of self-reinnervation and suggest that self-reinnervation can produce complete interconversion of muscle fiber types. Cross-reinnervation of FDL muscle by SOL motoneurons (SOL----FDL; n = 12) produced muscles that were smaller (about half the normal wet weight) and more red than normal. SOL----FDL muscle contracted more slowly than normal or FDL----FDL muscles and had much higher proportions of histochemical type I muscle fibers. In those SOL----FDL muscles, in which little or no unwanted self-reinnervation could be demonstrated, greater than 95% of the muscle fibers were type I. Forty-one individual motor units in SOL----FDL muscles were isolated by intracellular penetration in functionally identified SOL alpha-motoneurons. Their muscle units were all type S by physiological criteria (absence of "sag" in unfused tetani and marked resistance to fatigue). SOL----FDL muscle units had contraction times and fatigue properties that were essentially identical to those of type S units in the normal FDL. All of the seven units, successfully studied by glycogen depletion, exhibited histochemical type I fibers. SOL motoneurons that innervated FDL muscle units had slightly shorter afterhyperpolarization durations than normal SOL cells, but axonal conduction velocities were normal.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Carbonic anhydrase III in skeletal muscle fibers: an immunocytochemical and biochemical study.

1988 ◽  
Vol 36 (7) ◽  
pp. 775-782 ◽  
Author(s):  
P Frémont ◽  
P M Charest ◽  
C Côté ◽  
P A Rogers
Keyword(s):  
Skeletal Muscle ◽  
Carbonic Anhydrase ◽  
Vastus Lateralis ◽  
Muscle Fibers ◽  
Water Soluble ◽  
Type I ◽  
Fiber Types ◽  
Thin Sections ◽  
Carbonic Anhydrase Iii ◽  
Skeletal Muscle Fibers

The objectives of the present study were to determine if carbonic anhydrase III (CA III) demonstrated a specific association for any particular organelle or structure of the skeletal muscle cell and to quantify the activity and content of this enzyme in different types of skeletal muscle fibers. Ultrastructural localization of CA III in the soleus (SOL), deep vastus lateralis (DVL), and superficial vastus lateralis (SVL), composed of predominantly type I, IIa, and IIb fibers, respectively, was performed using a high-resolution immunocytochemical technique and antibody specific for CA III on ultra-thin sections of skeletal muscle embedded in the water-soluble medium polyvinyl alcohol (PVA). The results indicated a uniform distribution of CA III within the sarcomere. Mitochondria, nuclei, triads, Z-, and M-bands were not specifically labeled. Immunoblotting of washed myofibril preparations did not show any detectable CA III associated with this structure. In addition to quantification of the immunogold labeling, CA III activity and content were assayed in the post-mitochondrial supernatant of the three muscles. In the SOL, these values were found to be 3.6-7.6 times higher than in the DVL. The SVL showed a labeling intensity slightly higher than background level, while the enzyme activity and content were indistinguishable from background levels. We therefore conclude that CA III is randomly distributed in the cytoplasm of the three muscle fiber types and that the relative CA III content and activity in the three muscles studied is SOL greater than DVL greater than SVL approximately equal to 0.

scholarly journals Metabolic and morphometric profile of muscle fibers in chronic hemodialysis patients

2012 ◽  
Vol 112 (1) ◽  
pp. 72-78 ◽  
Author(s):  
Michael I. Lewis ◽  
Mario Fournier ◽  
Huiyuan Wang ◽  
Thomas W. Storer ◽  
Richard Casaburi ◽  
...  
Keyword(s):  
Muscle Fiber ◽  
Muscle Fibers ◽  
Exercise Intolerance ◽  
Maintenance Hemodialysis ◽  
Muscle Fiber Types ◽  
Vastus Lateralis Muscle ◽  
Type I ◽  
Fiber Types ◽  
Dense Matrix ◽  
Individual Fiber

Muscle weakness and effort intolerance are common in maintenance hemodialysis (MHD) patients. This study characterized morphometric, histochemical, and biochemical properties of limb muscle in MHD patients compared with controls (CTL) with similar age, gender, and ethnicity. Vastus lateralis muscle biopsies were obtained from 60 MHD patients, 1 day after dialysis, and from 21 CTL. Muscle fiber types and capillaries were identified immunohistochemically. Individual muscle fiber cross-sectional areas (CSA) were quantified. Individual fiber oxidative capacities were determined (microdensitometric assay) to measure succinate dehydrogenase (SDH) activity. Mean CSAs of type I, IIA, and IIX fibers were 33, 26, and 28% larger in MHD patients compared with CTL. SDH activities for type I, IIA, and IIX fibers were reduced by 29, 40, and 47%, respectively, in MHD. Capillary to fiber ratio was increased by 11% in MHD. The number of capillaries surrounding individual fiber types were also increased (type I: 9%; IIA: 10%; IIX: 23%) in MHD patients. However, capillary density (capillaries per unit muscle fiber area) was reduced by 34% in MHD patients, compared with CTL. Ultrastuctural analysis revealed swollen mitochondria with dense matrix in MHD patients. These results highlight impaired oxidative capacity and capillarity in MHD patients. This would be expected to impair energy production as well as substrate and oxygen delivery and exchange and contribute to exercise intolerance. The enlarged CSA of muscle fibers may, in part, be accounted for by edema. We speculate that these changes contribute to reduce limb strength in MHD patients by reducing specific force.

Metabolic capacity of individual muscle fibers from different anatomic locations.

1992 ◽  
Vol 40 (6) ◽  
pp. 819-825 ◽  
Author(s):  
B W Rosser ◽  
B J Norris ◽  
P M Nemeth
Keyword(s):  
Biceps Brachii ◽  
Muscle Fibers ◽  
Oxidative Capacity ◽  
High Energy ◽  
Triceps Surae ◽  
Type I ◽  
Fiber Types ◽  
Metabolic Capacity ◽  
The Individual ◽  
Anatomic Locations

We studied muscle fibers by quantitative biochemistry to determine whether metabolic capacity varied among fibers of a given type as a function of their anatomic location. Muscles were selected from both contiguous and diverse anatomic regions within the rats studied. The individual fibers, classified into myosin ATPase fiber types by histochemical means, were assessed for fiber diameters and analyzed for the activities of enzymes representing major energy pathways: malate dehydrogenase (MDH, oxidative), lactate dehydrogenase (LDH, glycolytic), and adenylokinase (AK, high-energy phosphate metabolism). We found that neither the average activities of each of the three enzymes nor the fiber diameters varied in Type I or Type IIa fibers selected from superficial to deep portions of the triceps surae of the hindlimb. However, the IIb fibers in the deep region of this muscle group had significantly greater oxidative capacity, less glycolytic capacity, and smaller diameters than the superficially situated IIb fibers. Type IIa fibers in lateral gastrocnemius, extensor digitorum longus, psoas, diaphragm, biceps brachii, superficial masseter, and superior rectus muscles were highly variable in both diameter and enzyme profiles, with a correlation between MDH activity and fiber diameter. Therefore, our results show that both intermuscular and intramuscular metabolic variations exist in muscle fibers of a given type.

scholarly journals Hemolymph supply to locomotor muscles of the ghost crab, Ocypode quadrata

2021 ◽  
Author(s):  
Siyuan Yang ◽  
Tera D. Douglas ◽  
Ryan Ruia ◽  
Scott Medler
Keyword(s):  
Blood Vessels ◽  
Skeletal Muscles ◽  
Muscle Fibers ◽  
Circulatory System ◽  
Histochemical Staining ◽  
Functional Coupling ◽  
Fiber Types ◽  
Ghost Crab ◽  
Ghost Crabs ◽  
Land Crabs

Ghost crabs are the fastest and most aerobically fit of the land crabs. The exceptional locomotory capacity of these invertebrate athletes seemingly depends upon effective coupling between the cardiovascular system and skeletal muscles, but how these systems are integrated has not been well defined. In the current study, we investigated the relationship between aerobic muscle fibers within the skeletal muscles used to power running and the blood vessels supplying these muscles. We used histochemical staining techniques to identify aerobic versus glycolytic fibers and to characterize membrane invaginations within the aerobic fibers. We also determined how the diameters of these two fiber types scale as a function of body size, across two orders of magnitude. Vascular casts were made of the blood vessels perfusing these muscles and special attention was given to small, capillary-like vessels supplying the fibers. Finally, we injected fluorescent microspheres into the hearts of living crabs and tracked their deposition into different muscle regions to quantify relative hemolymph flow to metabolic fiber types. Collectively, these analyses demonstrate that ghost crab muscles are endowed with an extensive arterial hemolymph supply. Moreover, the hemolymph flow to aerobic fibers is significantly greater than to glycolytic fibers within the same muscles. Aerobic fibers are increasingly subdivided by membrane invaginations as crabs increase in size, keeping the diffusive distances relatively constant. These findings support a functional coupling between a well-developed circulatory system and metabolically active muscle fibers in these invertebrates.

scholarly journals Ractopamine-induced fiber type-specific gene expression in porcine skeletal muscles is independent of growth

2020 ◽  
Vol 98 (11) ◽  
Author(s):  
Andrea M Gunawan ◽  
Con-Ning Yen ◽  
Brian T Richert ◽  
Allan P Schinckel ◽  
Alan L Grant ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscles ◽  
Muscle Hypertrophy ◽  
Citrate Synthase ◽  
Fiber Type ◽  
Muscle Growth ◽  
Specific Gene ◽  
Type I ◽  
Fiber Types ◽  
Adequate Diet

Abstract Feeding ractopamine (RAC), a β-adrenergic agonist (BAA), to pigs increases type IIB muscle fiber type-specific protein and mRNA expression. However, increases in the abundance of these fast-twitch fiber types occur with other forms of muscle hypertrophy and thus BAA-induced changes in myosin heavy chain (MyHC) composition may simply be associated with increased muscle growth known to occur in response to BAA feeding. The objective of this study was to determine whether RAC feeding could change the MyHC gene expression in the absence of maximal muscle growth. Pigs were fed either an adequate diet that supported maximal muscle hypertrophy or a low nutrient diet that limited muscle growth. RAC was included in diets at 0 or 20 mg/kg for 1, 2, or 4 wk. Backfat depth was less (P < 0.05) in pigs fed the low nutrient diet compared with the adequate diet but was not affected by RAC. Loin eye area was greater (P < 0.05) in pigs fed an adequate diet plus RAC at 1 wk but did not differ among remaining pigs. At 2 and 4 wk, however, pigs fed the adequate diet had greater loin eye areas (P < 0.05) than pigs fed the low nutrient diet regardless of RAC feeding. Gene expression of the MyHC isoforms, I, IIA, IIX, and IIB, as well as glycogen synthase, citrate synthase, β 1-adrenergic receptor (AR), and β 2-AR were determined in longissimus dorsi (LD) and red (RST) and white (WST) portions of the semitendinosus muscles. MyHC type I gene expression was not altered by RAC or diet. Feeding RAC decreased (P < 0.01) MyHC type IIA gene expression in all muscles, but to a greater extent in WST and LD. MyHC type IIX gene expression was lower (P < 0.05) in WST and LD muscles in response to RAC but was not altered in RST muscles. RAC increased (P < 0.05) MyHC type IIB gene expression in all muscles, but to a greater extent in RST. β 1-AR gene expression was unaffected by RAC or diet, whereas the expression of the β 2-AR gene was decreased (P < 0.001) by RAC. No significant RAC * diet interactions were observed in gene expression in this study, indicating that RAC altered MyHC and β 2-AR gene expression in porcine skeletal muscles independent of growth.

External and Internal Influences on Muscle Pathology

2012 ◽  
Vol 136 (8) ◽  
pp. 927-934 ◽  
Author(s):  
Paul E. McKeever ◽  
Sandra Camelo-Piragua ◽  
James Dowling
Keyword(s):  
Muscle Biopsy ◽  
Motor Nerve ◽  
Muscle Fibers ◽  
Neuromuscular Diseases ◽  
Muscle Pathology ◽  
Etiologic Factor ◽  
Fiber Types ◽  
Intrinsic Muscle ◽  
Histologic Pattern ◽  
Cellular Necrosis

Three cases of different types of neuromuscular diseases demonstrate different muscle responses to external stress or intrinsic muscle abnormalities. The first muscle biopsy shows stenosis of its vessels causing acute muscle ischemia, stress from an external vascular disease. The muscle response is similar to the cellular necrosis seen in primary muscle diseases (myopathies), but the histologic pattern is more focal than most myopathies. The second muscle biopsy demonstrates the effects of external motor nerve injury or disease causing groups of muscle fibers to atrophy. If a nerve reinnervates the muscle, it changes the fiber types in distinct patterns. The third muscle biopsy shows an intrinsic muscle abnormality causes chronic failure of the muscle fibers to thrive and repeated attempts by the fibers to regenerate, stimulating other tissue repair processes, like fibrosis, to change the muscle. Depending on the etiologic factor, muscle will respond to internal and external influences in different manners.

Absence of regional differences in the size and oxidative capacity of diaphragm muscle fibers

1987 ◽  
Vol 63 (3) ◽  
pp. 1076-1082 ◽  
Author(s):  
G. C. Sieck ◽  
R. D. Sacks ◽  
C. E. Blanco
Keyword(s):  
Muscle Fibers ◽  
Continuous Distribution ◽  
Oxidative Capacity ◽  
Cross Sectional Area ◽  
Diaphragm Muscle ◽  
Type I ◽  
Fiber Types ◽  
Sectional Area ◽  
Cross Sectional ◽  
Fast Twitch

The oxidative capacity and cross-sectional area of muscle fibers were compared between the costal and crural regions of the cat diaphragm and across the abdominal-thoracic extent of the muscle. Succinate dehydrogenase (SDH) activity of individual fibers was quantified using a microphotometric procedure implemented on an image-processing system. In both costal and crural regions, population distributions of SDH activities were unimodal for both type I and II fibers. The continuous distribution of SDH activities for type II fibers indicated that no clear threshold exists for the subclassification of fibers based on differences in oxidative capacity (e.g., the classification of fast-twitch glycolytic and fast-twitch oxidative glycolytic fiber types). No differences in either SDH activity or cross-sectional area were noted between fiber populations of the costal and crural regions. Differences in SDH activity and cross-sectional area were noted, however, between fiber populations located on the abdominal and thoracic sides of the costal region. Both type I and II fibers on the abdominal side of the costal diaphragm were larger and more oxidative than comparable fibers on the thoracic side.

Oxidative capacity, blood flow, and capillarity of skeletal muscles

1980 ◽  
Vol 49 (4) ◽  
pp. 627-633 ◽  
Author(s):  
L. C. Maxwell ◽  
T. P. White ◽  
J. A. Faulkner
Keyword(s):  
Blood Flow ◽  
Skeletal Muscles ◽  
Muscle Blood Flow ◽  
Muscle Fibers ◽  
Capillary Density ◽  
Oxidative Capacity ◽  
Histochemical Demonstration ◽  
Capillary Endothelium ◽  
Fiber Types ◽  
Fiber Area

Our purpose was to test the hypothesis that the capillarity of mammalian skeletal muscles is correlated with the oxidate capacity of muscle fibers, or with the capacity for maximum blood flow. Capillarity of skeletal muscles from several species was determined using histochemical demonstration of phosphatase activity of capillary endothelium. Serial sections were incubated for succinate dehydrogenase activity as an indicator of muscle fiber oxidative capacity, and for myofibrillar ATPase activity. three types of muscle fibers were identified. Fiber area was determined by planimetry of projected cross sections. Succinate oxidase activity of whole homogenates was determined by differential respirometry. Muscle blood flow was determined experimentally or data were obtained from the literature. No consistent relation was observed for the different fiber types in the number of adjacent capillaries. Capillary density was negatively correlated with mean fiber area. Among adult animals of several species, skeletal muscles representing a 17-fold range of oxidative capacity demonstrated no relation between capillarity and oxidative capacity or muscle blood flow at maximum oxygen uptake. We find no support for relations between oxidative capacity of muscle blood flow and the capillarity of whole muscle or individual fibers and reject the hypothesis.

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