Muscle fibre characteristics of active and inactive standardbred horses

Muscle biopsies were taken from the middle gluteus muscle of 36 Standardbred horses. Twelve of the horses were inactive, while 24 were actively trained and raced. Twelve of the trained horses were moderate performers, with a mean racing time of 1 min 21 secs per km (741 m/min) and the other 12 were excellent performers, with a mean racing time of 1 min 16 secs per km (789 m/min). The percentage and mean area of Type I fibres were similar in all three groups of horses. Marked differences were found among the subgroups of Type II fibres. The well-trained horses had a higher proportion of Type IIA fibres and a lower proportion of the Type IIB (58 per cent IIA and 15 per cent IIB fibres in the excellent performers; 49 per cent IIA and 26 per cent IIB in the moderate performers) than the inactive horses (41 per cent IIA and 35 per cent IIB fibres). The mean area of Type II fibres was smaller in the muscle of the active horses (excellent performers: IIA, 3075 micron2; IIB, 3378 micron2) moderate performers: IIA, 3185 micron2; IIB, 4252 micron2) than in that of the inactive ones (IIA, 3714 micron2; IIB, 5935 micron2). Intramuscular substrates (glycogen and triglycerides) and the activities of enzymes used as markers for the glycolytic potential of the muscle seemed to be similar in the three groups of horses, although there were large interindividual variation.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of testosterone on gastrocnemius muscle fibres in growing and adult male and female rats: a histochemical,morphometric and ultrastructural study

In this study, the effect of testosterone on gastrocnemius muscle fibres in growing and adult rats (male and female) was examined using histochemical, morphometric and ultrastructural techniques. After physiological saline (PS), olive oil (OvO) or olive oil + testosterone (OvOT) injections on 72 rats (growing and mature, 36 male and 36 female), the sample tissues of fibre types of the gastrocnemius muscle taken were examined by histochemical [alkaline adenosine triphosphatase (alk-ATPase), acid ATPase (ac-ATPase)], morphometric and ultrastructural techniques. In PS-injected control groups, the gastrocnemius muscle of both sexes contained all the fibre types studied [slow-oxidative muscle fibres (type I), fast-oxidative glycolytic muscle fibres (type IIA) and fast-glycolytic muscle fibres (type IIB)]. The type I fibres had the smallest diameter, type IIA had a medium diameter and type IIB fibres had the largest diameter. In OvO-injected groups, it was observed that the OvO had little effect on the gastrocnemius muscles of either sex, although there was significant enlargement of type IIB fibres. After the injection of OvOT, hypertrophy of muscle fibres was determined by morphometric study. The biggest increase in diameter was on type I fibres. In addition, degenerations on some mitochondria, accumulation of lipid droplets on type I and type II fibres, an increase in glycogen particles, bifurcation of myofibrils, an increase in the number and diameter of units resembling T tubules and an increase in ribosomal content were also observed in the same group by transmission electron microscope. Consequently, it was determined that testosterone can induce protein synthesis in gastrocnemius muscle fibres, and induces changes in shape and size, and also can change the appearance and the number of fibres.     

Estudio Histoquímico y Morfométrico de la Población Fibrilar del Músculo Diafragma. Parte I: Caballo y Perro

Histochemical and morphometric study of fibrilar population in diaphragm muscle. Part I: Horse and Dog By using m-ATPase techniques, under alkaline and acid pre-incubations, as well as metabolic techniques (α-MGPDH and NADH-TR), sections from the lumbar and costal portions of diaphragm muscle in horse and dog were analysed. Fibre samples were exposed to image-analysis observations in order to determine the lowest fibre diameter and obtain estadistical data. The findings revealed that the lumbar portion of horse diaphragm consists of fibre types I, IIA and IIB; in the costal portion, the fibre types I and IIA were present. In the dog, the lumbar and costal portions consisted of fibre types I, IIA and IIC, which showed some specific histochemical characteristics. Some data concerning both morphometrical and statistical fibre-type analysis are presented.     

Type,diameter and distribution of fibres in some respiratory and abdominal muscles of the goat

This study was designed to determine the histochemical properties, size and composition of fibres in the diaphragm, intercostal and abdominal muscles of goats to clarify whether reported similarities in respiratory muscle physiology between goats and humans have a structural basis. Serial sections (10 µm) of muscular tissue from adult female goats were stained for myosin adenosine triphosphatase and reduced nicotinamide adenine dinucleotide dehydrogenase-tetrazolium reductase activities; the fibres were classified into types I, IIA and IIB; and their mean diameter and composition were determined. Abdominal and intercostal muscles contained types I, IIA and IIB fibres in the ratio 1:1:1, and the mean diameter of the fibres ranged from 49.2 to 62.2 µm. In contrast, the diaphragm contained 58.9% type I and 41.1% type II fibres, and the latter could not be differentiated into types IIA and IIB. Diaphragmatic fibres were also smaller (36.9 – 40.9 µm). These findings contrast with those in humans, where the diaphragm, intercostal and abdominal muscles contain >50% type I fibres and have fibres of identical diameter. The differences in fibre characteristics between the diaphragm, intercostal and abdominal muscles of goats and the differences between goats and humans need to be taken into consideration in interpreting the results from studies in respiratory muscle physiology.Abbreviations mATPase myosin adenosine triphosphatase - NADH-TR nicotinamide adenine dinucleotide dehydrogenase-tetrazolium reductase     

The Post-larval Development of Lateral Musculature in Gilthead Sea Bream Sparus aurata (L.) and Sea Bass Dicentrarchus labrax (L.)

Fibre-type differentiation of lateral musculature has been studied in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.) during post-larval development using ultrastructural, histochemical and morphometric techniques. The study showed three muscle layers: red, intermediate (or pink) and white. Initially, most of the red muscle showed low myosin ATPase (m-ATPase) activity fibres, whereas near the transverse septum some small high m-ATPase activity fibres appeared and later acquired a rosette aspect. Afterwards, during adult growth the red muscle showed a histochemical mosaic appearance. The pink muscle in sea bass was observed at the beginning of juvenile development by the oxidative technique (NADH-RT reaction) whereas in gilthead sea bream it was also observed at the end of larval development. The pink layer consists of high m-ATPase activity fibres. However, along the muscle development other low and moderate m-ATPase activity fibres were observed close to the red and white muscles, respectively. The white muscle of juvenile fish showed a histochemical mosaic appearance near the pink muscle. In adult specimens the mosaic white muscle spread out occupying the whole of the myotome. Morphometric analysis shows a significant increase in mean fibre diameter during post-larval development, as shown by the Student's t-test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibres from the myosatellite cells (hyperplastic growth).     

Comparison of the histological and histochemical properties of skeletal muscles between carbon dioxide and electrically stunned chickens

1. Histological and histochemical profiles of Musculus pectoralis (PT, type IIB fibres), M. iliotibialis lateralis (ITL, types IIA + IIB fibres) and M. puboischiofemoralis pars medialis (PIF, type I fibres) were compared in carbon dioxide (37%, 70 s) and electrically (14 V, 5 s) stunned male chickens. 2. Muscle materials were taken at 0, 4 and 24 h from carcases dressed and cooled with ice-water mixture for 30 min. Glycogen and fat contents, and adenosine triphosphatase and reduced nicotinamide adenine dinucleotide dehydrogenase activities of fibres were measured. 3. In PT muscle at 0 h, gas stunned chickens showed many fibres with high glycogen content but those electrically stunned contained few such fibres. Fibres from gas stunned birds had lost almost all their glycogen after 24 h of cold storage. 4. In the ITL muscle of gas stunned chickens at 0 h residual glycogen was observed in type IIB fibres. In contrast, in the electrically stunned birds it was in type IIA, showing the different effects of the stunning methods. During cold storage, glycogen disappeared earlier in type IIB than IIA fibres. 5. In PIF muscle with fibres of low glycogen content, the gas stunned chickens maintained a good fibre structure for 4 h or more, but the electrically stunned had already lost intact fibre structure at 4 h. 6. These results indicated that the carbon dioxide stunning was a better method for chicken welfare and meat quality than electrical stunning.     

Energy metabolism in relation to skeletal muscle fibre properties during treadmill exercise

The skeletal muscle metabolism of horses with a wide range of cardiocirculatory capacities was studied during a standardised near-maximal exercise test in relation to muscle fibre composition. Although the same amount of work was performed by all horses the amount of triglycerides and glycogen utilised and lactate accumulated differed widely. Both blood and muscle lactate accumulation were positively related to the amount of glycogen utilised. These factors were in turn positively associated with the percentage of Type IIB fibres and consequently negatively associated with the percentage of Type IIA fibres. The percentage of Type IIB fibres was negatively correlated to the activities of the oxidative enzymes citrate synthase and 3-hydroxyacyl-CoA-dehydrogenase and positively associated with the lactate dehydrogenase activity. The results indicate that the horses with a high proportion of Type IIB fibres had a lower muscle oxidative capacity and were, therefore, for energy production, more dependent upon glycolysis with subsequent lactate accumulation than horses which had a high proportion of Type IIA fibres. The different metabolic properties of the muscle fibre types indicate that the pattern of muscle fibre recruitment could have a significant effect on intramuscular substrate utilisation.     

Oxidative capacity of skeletal muscle fibres in racehorses: histochemical versus biochemical analysis

The oxidative capacity of skeletal muscle fibre types was evaluated histochemically using the nicotinamide dinucleotide diaphorase (NADH-D) staining, and biochemically by measuring the activity of citrate synthase (CS) in both whole muscle samples and in pools of fibres of identified type. Duplicate determinations of the NADH-D staining pattern resulted in standard deviations (sd) between duplicates of 6 and 11 per cent for two observers. The NADH-D pattern was found to differ between observers. Duplicate determinations of CS activity in the same fibre pools resulted in an sd value of 2.9 mumol/g/min. Measurements of whole muscle CS activity did not provide information about the distribution of oxidative capacity among fibre types. The NADH-D stain and CS activity in fibre pools both showed that, in general, type I and IIA fibres had a higher oxidative capacity than type IIB fibres. Biochemical techniques also showed, however, that the CS activity in type I and IIA fibres of different horses could vary as much as twofold, whereas the NADH-D rating showed a high intensity staining for most type I and IIA fibres in all horses. Furthermore, type IIB fibres received a lower NADH-D rating than the other fibre types even when the CS activities were quite similar. For purposes of research, biochemical measurement of oxidative capacity in individual muscle fibre types provides valuable quantitative and comparative information. The ease of histochemical NADH-D staining in comparison to fibre dissections makes this technique more practical for routine estimates of the distribution of oxidative capacity among muscle fibres.(ABSTRACT TRUNCATED AT 250 WORDS)     

Muscle glycogen depletion patterns during draught work in Standardbred horses

Muscle fibre recruitment was investigated during draught loaded exercise by studying glycogen depletion patterns from histochemical stains of muscle biopsies from the gluteus and semitendinosus muscles. Three Standardbred trotters performed several intervals of draught loaded exercise on a treadmill with 34 kp at a trot (7 m/sec) and with 34 and 80 kp, respectively at a walk (2m/sec). Exercise was continued until the horses were unwilling to continue. Glycogen depletion was seen in all three fibre types when trotting with 34 kp for 5 or 10 mins. When an equal weight resistance was pulled at a walk, glycogen depletion was first seen in type I fibres only, then followed by a small percentage of type IIA fibres after at least 1 h. When 80 kp was pulled at a walk both type I and IIA fibres showed glycogen depletion, and after at least 30 mins exercise a small percentage of type IIB fibres was also depleted. These results indicate that the muscle fibres are depleted, in order, from type I through IIA to IIB as the intensity or duration of draught work increases.     

Histochemical properties of muscle fibres types and enzyme activities in skeletal muscles of Standardbred trotters of different ages

Fibre characteristics and enzyme activities were determined for the gluteus, semitendinosus, vastus lateralis and triceps brachii muscles of 55 Standardbred trotters of different ages. Four fibre types (I, IIA, IIB, IIC) were demonstrated by histochemical staining of myofibrillar adenosine triphosphatase after preincubation at different pH values. Type II fibres predominated in all the muscles and the type IIA/IIB ratio was higher in horses over 5 years than in younger horses, except in the vastus in which the IIA/IIB ratio did not change with age. The vastus had the highest proportion of type IIA fibres and the semitendinosus the highest proportion of type IIB fibres. Histochemical demonstration of NADH dehydrogenase disclosed that almost 100 per cent of the type IIA and many of the type I and IIB fibres were medium-stained; the remaining type I fibres were darkly stained and the type IIB fibres lightly stained. In older horses more fibres were stained for NADH dehydrogenase. The activity of triosephosphate dehydrogenase decreased that that of 3-hydroxy-acyl-coA dehydrogenase and citrate synthase increased in all the muscles except the vastus with increasing age. The greatest increase in oxidative capacity occurred in the gluteus and triceps. Training, rather than age, was regarded as the factor inducing these changes. The results emphasise that histochemical data are only semiquantitative, and there are apparent discrepancies in the intensities of histochemical staining and the biochemical evaluation of various enzymes.     

Age and exercise effects on mitochondrial density and capillary fibre ratio in bird leg muscle

1. Muscle fibre composition, capillary-fibre ratio and mitochondrial density of the Iliotibialis lateralis muscle were monitored during growth from the age of 8 weeks to sexual maturity (24 weeks) in sedentary birds and in birds subjected to regular treadmill exercise. 2. The Iliotibialis lateralis muscle consists almost exclusively of type IIA and IIB fibres with a small percentage of "unclassified" fibres. Over the 16-week period of growth there was no change in fibre composition but a marked fibre hypertrophy linked with a fall in the capillary-fibre ratio. Mitochondrial volume density increased with age but this was not significant. 3. Exercise significantly increased the IIA:IIB fibres ratio from 0.77 to 1.1 but had no significant effect on any other measured characteristics. 4. It is concluded that exercise at the intensity and duration employed (10-15 min daily, ca. 70-80% maximum oxygen consumption) has little effect on the growth and development of avian skeletal muscle although there was evidence of an increased oxidative capacity of the muscle.     

Enzyme-Histochemical Profiles of Fiber Types in Mature Canine Appendicular Muscles

The histochemical characteristics of skeletal muscle were assessed using a range of samples from 7 appendicular muscles taken from adult mixed-breed dogs (1.5 to 3 years of age). Two slow-twitch fiber subtypes (IA and IB) and three II subtypes (IIA, IIB and IIC) were identified according to myofibrillar myosin adenosine triphosphatase reaction after acid and alkaline preincubation. Type IIB fibers were not found in all muscles, and were only biologically significant in m. semiten-dinosus. The metabolic potential of these fibers is fairly similar to that of IIA fibers, but significantly different to that of IIB fibers in other mammals, suggesting that they may be designed to play a different functional role during locomotion. All canine muscle fibers have moderate to high oxidative capacity, which may be related to the extraordinary athletic capability of the species.     

Histochemical and morphometric aspects of the lateral musculature of different species of teleost marine fish of the Percomorphi order

Cross-sections of the lateral musculature of 19 species of teleost fish of the Percomorphi order were studied histochemically and morphometrically. The red, pink and white muscles were observed in all the species except in Seriola dumerilii and Scomber japonicus, in which no pink muscle was found. The red fibres were the smallest due to their minimal diameters. Histochemically and according to their location at least five types were found in the same species: typical red, in rosette, of the border, infiltrated pink and in mosaic. The pink fibres were larger than the red but smaller than the white. The form and disposition of the pink muscle varied according to the species. This muscle is made up by fibres of medium size, moderate m-ATPase and intermediate oxidative activities. Their histochemical properties change near red and white muscles, where they acquire the histochemical characteristics of the typical red fibres or the low m-ATPase activity of white fibres, respectively. The white muscle is the widest muscular layer. All of the species, except Thalassoma pavo are made up by three fibre types (small with high m-ATPase activity, medium with moderate m-ATPase activity, and large with low m-ATPase activity). They are mixed and show a histochemical and morphometric mosaic form. The extent of this mosaic depends on the species. In most of the analysed species it was observed in areas close to the pink muscle, but in three of them (Liza aurata, Seriola dumerilii and Coris julis) it occupied all of the myotome.     

Glycogen depletion patterns in horses performing maximal exercise

Muscle biopsy samples were collected from the left middle gluteal muscle of horses participating in competitive barrier trials. Twelve horses were biopsied the day before and within 30 minutes of completion of an 800 m barrier trial. A further six horses were sampled the day before, and within 30 minutes of, completion of a 1200 m barrier trial. Serial muscle sections were examined histochemically for myosin adenosine triphosphatase activity after acid preincubation, to demonstrate type I, IIA and IIB fibres. The glycogen content in the individual fibres was assessed using the periodic acid Schiff (PAS) reaction. Total glycogen in muscle was measured fluorimetrically after hydrolysis to glucose. Significant decreases in total muscle glycogen levels of 167.1 +/- 22.2 and 158.1 +/- 23.2 mmoles glucose units kg-1 (dry weight) (mean +/- standard error) occurred from pre- to post trial samples for horses in both 800 and 1200 m trials respectively. There were no significant differences between the decreases in total muscle glycogen from horses in either 800 m or 1200 m trials. In both 800 m and 1200 m trials there was a significant decrease in the percentage of type IIA and IIB fibres classified as having high PAS staining intensity, and a significant increase in the percentage of fibres classified as having medium staining intensity when samples taken after the trial were compared to those taken before the trial.     

Morphometry and histochemistry of the semitendinosus muscle of tufted capuchin monkeys (Cebus apella Linnaeus, 1758)

Considering monkeys are animals closely related to the human, and semitendinosus muscle has been used in many postural research experiments, we have decided to study its histochemical characteristics. Samples were removed from the proximal, middle and distal regions of the semitendinosus muscles of five adult male tufted capuchin monkeys and observed for reaction with m-ATPase (with alkaline and acid pre-incubation), nicotinamide adenine dinucleotide tetrazolium reductase, and haematoxylin and eosin. The muscle fibres were classified as fast glycolytic (FG), fast oxidative glycolytic (FOG) and slow oxidative (SO) and quantified in terms of frequency and area. The three regions of the semitendinosus muscle showed no significant differences in frequency or area of the respective fibre types, and therefore the muscle can be considered histoenzymologically homogeneous. FG fibres presented higher frequencies and larger areas. The sum of FOG and SO fibres was 57.5%, suggesting that the semitendinosus muscle of the tufted capuchin monkey is adapted to an action involved in posture maintenance and in long arboreal dislocation. The present data agrees with the notion of differentiated quadrupedalism in some primates, which support a lower percentage of their weight on the fore limbs.     

Lateral musculature in the whitemouth croaker (Micropogonias furnieri): its characterization with respect to different gonadal conditions

Histochemical and ultrastructural studies were performed on the lateral musculature from individual female whitemouth croaker, Micropogonias furnieri, at the anterior, medium and posterior regions. Based upon histochemical myosin-ATPase (m-ATPase) determination, diverse types of red, pink and white fibres were discerned. Red muscle had abundant mitochondria and stained intensely for aerobic enzymes, white muscle scarcely stained for the same enzymes and pink muscle responded in an intermediate manner. Both white and pink muscle had few mitochondria. The relative proportion of red muscle increased towards the caudal region; pink muscle diminished towards this region and white muscle modified its proportion only in the anterior region. m-ATPase activity showed differences in relation to the gonadal condition along the body, particularly in the white fibres at the anterior and medium regions.     

Muscle fibre type composition in untrained and endurance-trained Andalusian and Arab horses

Muscle biopsies were taken from the middle gluteal muscle of 68 stallions (52 Andalusian [AN] and 16 Arab [AR]) ranging from six to 12 years of age. Seventeen AN horses and eight AR horses were untrained, while the remainder underwent active endurance training for 6 months. Fifteen AN horses were moderately endurance-trained while the other 20 AN horses and eight AR horses were strongly endurance-trained. Percentages of type I and type II fibres were similar in all groups (P greater than 0.05). The group of horses with the hardest training had a higher percentage of type IIA fibres (AN P less than 0.01; AR P less than 0.02) and a lower percentage of type IIB fibres (P less than 0.05 for AN and AR) than the untrained horses. All trained horses had a higher percentage of type IIB oxidative fibres and a lower percentage of type IIB non-oxidative fibres than the untrained horses (P less than 0.01 for both). These results suggest that the proportions of type I and type II fibres are highly stable within a given breed, but the stimulus of training facilitates changes both in contractile properties and, particularly, in the oxidative capacity of type II subgroups.     

Skeletal muscle characteristics in young trained and untrained standardbred trotters.

Muscle biopsies were taken from the middle gluteal muscle of 28 Standardbred trotters, 3-4 years of age. The 13 horses in Group T were trained consistently from 18 months of age, whereas the 15 horses in Group UT were not exposed to any systematic training before 3 years of age. Group T horses had a lower percentage of Type IIB fibres (31%) than did Group UT horses (39%). Citrate synthase (CS) activity, representing oxidative capacity, was higher in Group T (72 mmol kg-1 min-1) than in Group UT (47 mmol kg-1 min-1). Biopsies were taken from 4 horses in each group when they were foals and then annually until 3-4 years of age. Results from this study indicate that regular training of Standardbreds from 18 months of age resulted in increased CS activity and a decrease in the percentage of Type IIB fibres. This study shows that training, not growth, is the main factor that induces a high oxidative capacity and a high Type IIA/IIB fibre ratio in muscle of Standardbred trotters.     

Carnosine, anserine and taurine contents in individual fibres from the middle gluteal muscle of the camel

High muscle camosine and anserine contents contribute significantly to intra-cellular physico-chemical buffering. Our aim was to measure carnosine, anserine and taurine contents directly in individual type I, HA and IIB fibres from the middle gluteus muscle of the camel. Mean carnosine contents in type I, IIA and IIB were 24·6 ±9·2, 39·4 ±11·4 and 42·8 ±18·8 mmol kg⁻¹ dry weight (dw), respectively. Mean anserine contents in type I, HA and IIB fibres were 30·0 ±8·4, 37·3 ±10·1 and 34·5 ±9·7 mmol kg⁻¹ dw, respectively. Mean taurine contents in type I, IIA and HB fibres were 42·4 ±15-9, 203 ±12·9 and 24·7 ±15·9 mmol kg⁻¹ dw, respectively. Higher carnosine contents in type II fibres emphasise the importance of carnosine to intra-muscular acid-base regulation. A specific role for taurine in type I fibres is unclear.     

Growth- and breed-related changes of muscle fiber characteristics in cattle

The objective of this study was to investigate the growth- and breed-related changes of muscle fiber characteristics in cattle and their importance to meat quality. Four cattle breeds with different growth impetus and muscularity were reared and slaughtered under experimental conditions. German Angus as a beef type, Galloway as a hardy type, Holstein Friesian as a dairy type, and double-muscled Belgian Blue as an extreme type for muscle growth were used. Between 5 and 17 bulls of each breed were slaughtered at 0, 2, 4, 6, 12, 18, and 24 mo of age. Muscle fiber traits were determined and classified by computerized image analysis, and several measures of meat quality were also determined, including shear force value, meat color, and i.m. fat content. The postnatal growth of semitendinosus muscle in cattle was characterized by a nearly 10-fold increase of muscle fiber area from birth to 24 mo of age. In the first few months after birth, a transformation of type IIA fibers into IIB fibers was found, whereas type I fibers were nearly unaffected by age. The apparent total muscle fiber number of semitendinosus muscle did not increase during postnatal life. These results confirm that the fiber number is determined in embryonic development. Throughout the study, the double-muscled Belgian Blue (BBDM) bulls had almost twice the fiber number of the other breeds, emphasizing a more extensive hyperplasia of muscle fibers during embryonic development in BBDM compared with the other three breeds. The apparent number of type I fibers was, however, not affected by breed, which suggests that the additional fibers found in BBDM postnatally were type IIB and IIA fibers. We did not find significant differences in muscle fiber total number, muscle fiber type frequencies, or meat quality characteristics among breeds, with the exception of BBDM. Having pooled the four breeds, paler meat was related to a higher frequency of type IIB fibers, a lower area of type IIA and type I fiber, and a higher total muscle fiber number. These findings based on data of double muscling give us some hints for biological causes for the variation of meat quality. Further investigation, in particular within each breed, is necessary to identify the superior fiber traits for bovine meat production.