Fibre types in the locomotory muscles of an antarctic teleost,Notothenia rossii

Summary The metabolic and structural differentiation of locomotory muscles of Notothenia rossii has been investigated. In this species sustained locomotion is achieved by sculling with enlarged pectoral fins (labriform locomotion), whilst the segmental myotomal muscle is reserved for burst activity. Red, white and subepidermal fibres can be distinguished in the trunk by histochemical and ultrastructural criteria. The main pectoral muscle (m. adductor profundus) consists entirely of red fibres. These three main fibres types show differences in histochemical staining profiles, capillarization, myofibril shape and packing, and lipid and mitochondrial content. The fractional volume of mitochondria amounts to 38% for pectoral, 30% for red myotomal and 1.9% for white myotomal fibres. Enzyme activities of red pectoral muscle are consistent with a higher potential for aerobic glucose and fatty acid oxidation than for the red myotomal fibres. Mg²⁺ Ca²⁺ -myofibrillar ATPase activities are similar for red pectoral and myotomal muscles and approximately half of those white fibres. Specialisations of N. rossii muscles associated with labriform swimming and locomotion at Antarctic temperatures are discussed.     

A histochemical study of the lateral muscles of five teleost species

A qualitative histochemical study has been made of the myotomal muscles of five teleost fish (glass fish, Chanda ranga; carp, Carassius carassius; coalfish, Gadus virens; black mollie, Molliensia sp. and grey mullet, Mugil cephalus ) . Three or four main fibre types were distinguished in these species on the basis of the distribution and relative activities of glycogen, lipid, aglycerophosphate dehydrogenase, phosphorylase, and succinic dehydrogenase. The so-called red and white fibre types were found to have similar histochemical properties to previously investigated species. All the species studied, with the exception of the glass fish, Chanda ranga , were found to have one or two types of pink fibre situated between the red and white fibre regions. In the carp, coalfish and mullet, the pink fibres were found to be composed of small and large diameter fibres which were similar to red and white fibres respectively, except for their staining for succinic dehydrogenase. Considerable differences were found in the relative amounts of pink muscles between species. Minor fibre components were found in several species. These consisted of very small diameter fibres which did not stain well with any of the histochemical procedures used. It is suggested that these fibres represent areas of continuing muscle growth. The results obtained are discussed in relation to the division of labour between myotomal muscles during swimming.     

Growth dynamics of caudal and pectoral fin muscle fibres in a carangid, Caranx malabaricus (Cuv. & Val.), and their possible relation with somatic growth

The growth dynamics of red, pink and white fibres of the caudal and pectoral fin muscles are described in Carans malabaricus (Cuv. & Val.) in relation to their somatic growth. In all three fibre types growth occurred by an increase in fibre number and diameter in small size classes of fish and by an increase in diameter only in larger fish. The growth dynamics of the three fibre types were similar to those of the myotomal muscle fibres and paralleled the somatic growth pattern of this fish.     

The distribution of capillaries in the somatic musculature of two vertebrate types with particular reference to teleost fish

The distribution of capillaries in teleost and rat striated muscles was investigated using a number of different methods. A new method for directly viewing capillaries was developed. Teleost white muscle has a capillary: fibre (C:F) ratio of between 0.2 and 0.3; and 0.6 to 1.0 peripheral capillaries per muscle fibre. 26-49% of fibres had no peripheral capillaries. Values for the rat gastrocnemius were 1.2, 2.6 and 4.8% respectively which compares well with literature values. Flathead red muscle had a C:F ratio of between 1.9 and 2.5; and between 5.3 and 6.6 peripheral capillaries per muscle fibre depending on the method used. Values for rat soleus were 1.8 and 4.1 respectively. Teleost pink fibres had an intermediate number of capillaries. Rat striated muscle, particularly the gastrocnemius, was found to be heterogeneous with respect to the distribution of capillaries. Flathead red muscle was homogeneous whilst teleost white muscle was only slightly variable. Flathead red muscle fibres are well suppled with subsarcolemmal mitochondria. These show a clumped distribution corresponding to the position of capillaries. In contrast teleost white fibres are almost totally devoid of these and all other mitochondria. No differences were observed in the vascularisation of either muscle type along the length of the fish. The results are discussed in relation to the division of labour between fibre types during swimming.     

Histochemical characterization of myotomal muscle in the grass pickerel, Esox americanus vermiculatus (LeSeuer), and the muskellunge, E. masquinongy (Mitchell)

A histochemical study of the myotomal muscles in the grass pickerel, Esox americanus vermiculatus , and the muskellunge, E. masquinongy , was performed using actomyosin ATPase and NADH diaphorase activities. Three fibre types, i.e., red, white and pink were distinguished on the basis of their enzyme activities. White muscle fibres comprised the bulk of the myotomal musculature. The relative proportion of red muscle fibres was greater in the caudal region than in more anterior regions of the body. Pink fibres formed only a few layers between red and white. These findings are discussed in relation to the possible functional significance of the muscle fibre types in swimming and feeding behaviour in these species.     

Histochemical characterization of myotomal muscle in the bluntnose minnow, Pimephales notatus Rafinesque

A histochemical study of the axial muscles in the bluntnose minnow, Pimephales notatus Rafinesque, revealed three main types of fibres-red, white and pink. These were distin- guished on the basis of glycogen or lipid contents, or of NADH diaphorase and actomyosin ATPase activities. White muscle formed the largest fraction of the total, whereas red ranged from slightly > 1% of the total cross-sectional area in the immediately post-pectoral fin region to slightly <4% in the post-anal region. Pink muscle formed only a few layers of fibres between red and white. Results are discussed in relation to possible division of labour among the different types of fibres. However, it is emphasized that fibre types may be more functionally versatile in fish muscles than has frequently been supposed.     

Histochemical fibre types in the lateral muscle of fishes in fresh, brackish and salt water

The histochemical pattern of red, pink and white muscle of fish living in fresh, brackish, and salt water is reported. The muscle fibres were stained routinely during the year for lactate dehydrogenase (LDH), menadione α-glycerophosphate dehydrogenase (Mα—GPDH), succinic dehydrogenase (SDH), myosin adenosine triphosphatase (myosin ATPase), phosphorylase, lipids and glycogen. The pink and red muscles contain more glycogen and lipids and have a higher SDH activity, which is in accord with their aerobic metabolism and function in sustained swimming activity. The acid labile myosin ATPase activity characteristic of fast twitch fibres is present in the white fibres of most species, however in the white muscle of Gobius paganellus the enzyme activity is stable to both acid and alkali and, in addition, there is a scattered distribution of different fibre types in red and, especially, pink muscle. A study of seasonal variation patterns of myosin ATPase in white muscle of mugilidae over a period of two years has demonstrated, in late summer, the appearance of new small diameter fibres, with a high acid stable enzyme activity, that develop into the large diameter acid labile fibres.     

Morphometry of muscle fibre types in the carp (Cyprinus carpio L.)

Summary Ultrastructural parameters of muscle fibre types of the carp (Cyprinus carpio L.) were measured and compared with their contractile properties. In red fibres, which are slower than pink fibres, the relative length of the junction between the T system and the sarcoplasmic reticulum (T-SR junction) is smaller and the Z lines are thicker than in pink fibres. Pink fibres have a smaller relative length of T-SR junction than white fibres from the axial muscles. The two types of red fibres present in carp muscle also differ in their relative lengths of T-SR junction. Significant differences in the relative areas of the SR were not found.The relative volume of myofibrils in red fibres is two-thirds that in pink fibres, a difference that is not reflected in the maximal isometric tetanic tensions of these types. Red fibres, which are less easily fatigued than pink fibres, have larger relative volumes of subsarcolemmal and intermyofibrillar mitochondria. Small pink fibres have a larger relative volume of subsarcolemmal mitochondria than large pink fibres, but have a similar relative volume of intermyofibrillar mitochondria. Small and large pink fibres differ in the relative volumes of their membrane systems, but have similar relative lengths of T-SR junction.     

The claw closer muscle of two estuarine crab species,Cyrtograpsus angulatus and Neohelice 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.     

The lateral musculature of the common bully, Gobiomorphus cotidianus, a freshwater fish from New Zealand

A histochemical and ultrastructural study has shown that the myotome of the common bully, Gobiomorphus cotidianus , is composed of three muscle fibre types: white, pink and small diameter fibres. There are no red fibres. Both white and pink fibres have characteristics similar to these fibres found in other teleosts. The small diameter fibres are located in the position usually occupied by red fibres and are identified by their small size and poor staining characteristics. At the ultrastructural level these small fibres are seen to have few mitochondria and a poorly developed sarcoplasmic reticulum. It is suggested that the small diameter fibres are a type of tonic muscle used for positioning the trunk.     

The ultrastructure and vascular supply of the different fibre types in the axial muscle of the sturgeon Acipenser stellatus,Pallas

Summary The ultrastructure and vascular supply of the different fibre types in the lateral muscles of the sturgeon Acipenser stellatus were studied by light- and electron microscopy and morphometry. Three fibre types form separate layers without intermingling. The red fibres are superficial, the white fibres deep and the intermediate fibres between them. From morphometric analyses, the mitochondrial volume fraction in red fibres is 30%, in intermediate fibres 3.7% and in white fibres 0.7%. Z lines are most fuzzy in the red fibres. Triads of the sarcotubular system are always situated at the level of the Z discs. In red fibres the three elements are arranged in a series along the myofibrils, whereas in white fibres they are arranged transversely and in the intermediate fibres they are aligned obliquely. The number of capillaries surrounding each fibre is 2.3, 0.9 and 0.2 for the red, intermediate and white fibres, respectively. In red fibres 16% of the surface is directly covered by capillaries. The corresponding percentages for intermediate and white fibres are 5 and 1, respectively. Per unit volume of the fibre, the directly vascularised fibre surface in red fibres is about ten times larger than that of white fibres.The degree of vascularisation of the fibre types is directly related to the volume fraction of mitochondria, and thus to their aerobic capacities.     

Morphological and histochemical characterization of the pectoral fin muscle of batoids

Batoids differ from other elasmobranch fishes in that they possess dorsoventrally flattened bodies with enlarged muscled pectoral fins. Most batoids also swim using either of two modes of locomotion: undulation or oscillation of the pectoral fins. In other elasmobranchs (e.g., sharks), the main locomotory muscle is located in the axial myotome; in contrast, the main locomotory muscle in batoids is found in the enlarged pectoral fins. The pectoral fin muscles of sharks have a simple structure, confined to the base of the fin; however, little to no data are available on the more complex musculature within the pectoral fins of batoids. Understanding the types of fibers and their arrangement within the pectoral fins may elucidate how batoid fishes are able to utilize such unique swimming modes. In the present study, histochemical methods including succinate dehydrogenase (SDH) and immunofluoresence were used to determine the different fiber types comprising these muscles in three batoid species: Atlantic stingray (Dasyatis sabina), ocellate river stingray (Potamotrygon motoro) and cownose ray (Rhinoptera bonasus). All three species had muscles comprised of two muscle fiber types (slow-red and fast-white). The undulatory species, D. sabina and P. motoro, had a larger proportion of fast-white muscle fibers compared to the oscillatory species, R. bonasus. The muscle fiber sizes were similar between each species, though generally smaller compared to the axial musculature in other elasmobranch fishes. These results suggest that batoid locomotion can be distinguished using muscle fiber type proportions. Undulatory species are more benthic with fast-white fibers allowing them to contract their muscles quickly, as a possible means of escape from potential predators. Oscillatory species are pelagic and are known to migrate long distances with muscles using slow-red fibers to aid in sustained swimming.     

Fibre types in locomotory muscles of the cartilaginous fish Chimaera monstrosa

The cartilaginous fish Chimaera monstrosa swims slowly by means of pectoral fin movements, and fast by undulations of the tail. In order to compare the fibres in the corresponding muscles, they were studied by histochemistry and electron microscopy. Three fibre types were identified by microphotometry and morphometry. Most of the axial muscles are white fibres, containing little mitochondria and glycogen. Red fibres, with glycogen and about 5 % mitochondria constitute a thin sheet in the axial muscles, composed of one fibre layer only. Pink fibres, with intermediate amounts of glycogen and mitochondria are situated between these two types, but are often not covered by red fibres. Pectoral muscles contain numerous red and intermediate fibres, partially mixed, superficially, and white fibres deeper. Pectoral muscle red fibres contain about 25 % mitochondria, half of which are situated in subsarcolemmal accummulations. The sarcotubular system has T-tubules at the Z discs, and the terminal cisternae are partially divided by regularly spaced clefts.     

Allometric dependences of oxygen transport parameters in skeletal muscles during ontogenesis of domestic fowl

In domestic fowl, from day 10 of embryogenesis to six month of postnatal life, investigation into the white glycolytic pectoral and red oxidative gastrocnemius muscles allometric dependences of the structural and functional parameters providing muscular fibres by oxygen: speed of breath of an isolated muscular fibre, size of a surface of an external membrane of mitochondria in a fibre, its permeability to oxygen, density of the capillaries located around of a fibre and in all muscle, volumetric speed of a muscular blood flow, and connection of speed of breath of muscular fibres with weight of a body of a bird.     

Muscle fibre types and size distribution in sub-antarctic notothenioid fishes

The presumptive tonic muscles fibres of Cottoperca gobio, Champsocephalus esox, Harpagifer bispinis, Eleginops maclovinus, Patagontothen tessellata, P. cornucola and Paranotothenia magellanica stained weakly or were unstained for glycogen, lipid, succinic dehydrogenase (SDHase) and myosin ATPase (mATPase) activity. Slow, intermediate and fast twitch muscle fibres, distinguished on the basis of the pH stability of their mATPases, showed intense, moderate and low staining activity for SDHase, respectively. Slow fibres were the major component of the pectoral fin adductor profundis muscle. The proportion of different muscle fibre types varied from the proximal to distal end of the muscle, but showed relatively little variation between species. The myotomes contained a lateral superficial strip of red muscle composed of presumptive tonic, slow twitch and intermediate fibres, thickening to a major wedge at the horizontal septum. All species also had characteristic secondary dorsal and ventral wedges of red muscle. The relative abundance and localization of muscle fibre types in the red muscle varied between species and with body size in the protandric hermaphrodite E. maclovinus. The frequency distribution of diameters for fast twitch muscle fibres, the major component of deep white muscle, was determined in fish of a range of body sizes. The absence of fibres <20 μm diameter was used as a criterion for the cessation of muscle fibre recruitment. Fibre recruitment had stopped in P. tessellata of 13·8 cm L_T and E. maclovinus of 32·8 cm L_T, equivalent to 49 and 36·5% of their recorded maximum sizes respectively. As a result in 20‐cm P. tessellata, the maximum fibre diameter was 300 μm and 36% of fibres were in excess of 200 μm. The unusually large maximum fibre diameter, the general arrangement of the red muscle layer and the extreme pH lability of the mATPase of fast twitch fibres are all common characters of the sub‐Antarctic and Antarctic Notothenioids, including Cottoperca gobio, the suggested sister group to the Notothenidae.     

Energy metabolism of carp swimming muscles

Summary Electromyography has been used to study the recruitment of red, pink and white muscle fibres of the Mirror carp at different swimming speeds. Locomotion below 0.3–0.5 L/S (lengths per second) is achieved primarily by fin movements after which the red myotomal muscle becomes active. Pink muscle fibres are the next type to be recruited at speeds around 1.1–1.5 L/S. White muscle is only used for fast cruising in excess of 2–2.5 L/S and during bursts of acceleration.Studies of the myofibrillar ATPase activities of these muscles have shown a ratio of 124 for the red, pink and white fibres respectively. The myosin low molecular weight components, which are characteristic of the myosin phenotype, have been investigated by SDS polyacrylamide electrophoresis. The light chain patterns of the pink and white muscles were identical and characteristic of the fast myosin phenotype. Red muscle myosin had a light chain pattern characteristic of slow muscles. It would appear that there is a relationship between the speed of contraction of the fibre types and the locomotory speed at which they are recruited.The activities of some enzymes of energy metabolism have also been determined in the three muscle types. Enzymes associated with oxidate metabolism have high, intermediate and low activities in the red, pink and white muscles respectively. Pyruvate kinase and lactate dehydrogenase activities were considerably higher in the pink than in either red or white muscles. It is suggested that the high capacity for anaerobic glycolysis of the pink muscle is associated with its recruitment for sustained effort at swimming speeds above which the fish can no longer meet all its energy requirements by gas exchange at the gills.Abbreviations used EDTA ethylenediamine tetraacetic acid - L/S lengths, sec^–1 - LDH Lactate dehydrogenase - PFK phosphofructokinase - SDS sodium dodecyl sulphate - TCA trichloroacetic acid     

Ultrastructure of muscle fibres in head and axial muscles of the perch (Perca fluviatilis L.)

Summary White, pink, red and deep red fibres, selected from a head muscle and from axial muscles of the perch, show significant differences in actin filament length, Z line thickness, Z line lattice space, myofibril girth, the percentages volume occupied by T system and terminal cisternae of the SR, and in the degree of T system SR contact per sarcomere. In both muscles the degree of T system SR contact decreases in the order: white, pink, red, deep red, which suggests a decrease of contraction velocity in the same order.The position of the T system (at the Z line or at the AI junction) is related to the actin filament length. The actin filaments in the red fibres are appreciably longer than in the white, which suggests that the sarcomeres of the red fibres have a broader length-tension curve. The Z line thickness is positively correlated with the actin filament length and, in the white and the red fibres, negatively with the degree of sarcomere shortening. Thicker Z lines are suggested to allow greater sarcomere sizes (length or girth).The percentage volume occupied by mitochondria varies independently of the extent of membrane systems.The ultrastructural characteristics of the fibre types are in agreement with the functional roles as reported in literature.     

Characterization of swimming muscle in the Atlantic mackerel, Scomber scombrus L.

Both red and white muscle were removed from juvenile and adult Atlantic mackerel, Scomber scombrus L., for histochemical characterization of the muscle fibre types. Staining of white muscle for myosin ATPase, SDH, NADH diaphorase, GPDH and LDH revealed that these fibres are homogeneous. Red muscle was shown to be heterogeneous, of at least two fibre types recognizable on the basis of myosin ATPase staining with preincubation at a pH of 9·8. These two red types are dispersed throughout the red muscle and are present in both juveniles and adults. Red muscle is located both deep within the myotomes and as a superficial layer of muscle fibres. A third group of muscle fibres, intermediate in nature, was distinguished at the apex of the red muscle 'triangle,' between the epaxial and hypaxial muscle, using NADH diaphorase and myosin ATPase stains. This paper discusses the possibility that functionally different muscle fibres occur in the red swimming muscle of the Atlantic mackerel.     

On the heterogeneity of capillaries of pigeon pectoralis muscle: a histoenzymatic and ultrastructural study

Synopsis Earlier studies had failed to show the presence of capillaries between the white fibres of pigeon pectoralis muscle. In this paper, data are reported for the first time documenting that these capillaries occur in both intra- and inter-fasicular areas of the muscle. Fresh frozen sections of pigeon pectoralis major muscle were incubated for alkaline ATPase reaction following pretreatment with different EDTA solutions (4.3 mM, pH 4.3). The results showed the existence of an inherent heterogeneity of capillaries. The capillaries of white fibres stained intensely for K^–/Mg^2–-EDTA or Mg²⁺-EDTA pre-incubated ATPase; the capillaries of red fibres stained poorly. Both white fibre and red fibre capillaries were examined ultrastructurally in the non-perfused pigeon pectoralis muscle. It is suggested that a possible correlation exists between the distinctive metabolic and mechanical characteristics of the Type II white, glycolytic, fast-twitch fast-fatigue muscle fibres and the high ATPase activity of their capillaries.     

AMP-deaminase in elasmobranch fish: a comparative histochemical and enzymatic study

AMP-deaminase activity was measured in white muscle from a wide range of fish, including one cyclostome, 13 chondrosteans, and one teleost to elucidate the pattern of the AMP-deaminase activity in white muscle of fish. Compared to a mammalian (rat) muscle extract, low enzyme activities are found in the cyclostome and two elasmobranchs from two families (Scyliorhinidae, Hexanchidae). In contrast, higher AMP-deaminase activities, similar to mammals, are expressed in Squalidae, all families of skates, Chimaeridae and in the teleostean fish. We then compared AMP-deaminase activities in red and white muscles from two representative elasmobranch fish, the dogfish (Scyliorhinus canicula) and the thornback ray (Raja clavata). The fibre type composition and distribution of the locomotory musculature were determined in these two elasmobranchs to establish a relationship between the morphology, the type of fibres of the locomotion-implicated muscles and the AMP-deaminase activity. Experimental data are discussed with respect to the layout of fibres in the myotome. In both species, three fibre types were identified. In the two fish myotomes, most of the axial muscles are white fibres while red fibres constitute a thin sheet. Some differences were observed between the two species in the distribution of intermediate fibres: in dogfish, these are located between the red and white fibres; in thornback ray, some are dispersed within the white fibre region, while others form an intermediary layer like in dogfish. These results suggest that in the course of evolution, an amplification of the AMP-deaminase activity in muscle was coupled with increase of complexity of the muscular structure.