A study of the swimming performance of the Crucian carp Carassius carassius (L.) in relation to the effects of exercise and recovery on biochemical changes in the myotomal muscles and liver

A study has been made of the maximum sustained swimming speed of Crucian carp Carassius carassius (L.) using a fixed velocity technique. The data obtained from swimming tests on 214 carp have been analysed using the method of probit analysis. The 50% fatigue level for 13–16 cm fish acclimated to 9.5±0.6°C has been estimated to be 3.35 lengths/sec. Biochemical measurements have been made on the red and white myotomal muscles and liver of fish subjected to both varying intensities of sustained swimming and short periods of vigorous swimming. Free creatine was found to increase only during high speed swimming in the white muscle. Elevated lactate concentrations occurred at both low and high sustained swimming speeds in the red superficial muscle but not during short periods of strenuous exercise. Glycogen depletion from the red musculature also only took place at the sustained swimming speeds investigated. The reverse situation was operative in the white muscle, significant glycogen depletion occurring only at the highest swimming speed studied. Lactate levels were only significantly different from non-exercised fish in the fish swimming at the higher velocities. The effects of periods of recovery following 200 min of sustained swimming were also investigated. White muscle lactate was at a higher level than non-exercise fish 5 h post-exercise, while both red muscle glycogen and lactate rapidly returned to pre-exercise concentrations. Biochemical measurements on the myotomal muscle types have been discussed in relation to the swimming performance of the fish and the division of labour between red and white fibres.     

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.     

Studies on the swimming musculature of the rainbow trout I. Fibre types

A histochemical study has been made on the myotomal musculature of the rainbow trout Salmo gairdneri Richardson. Four main types of fibre can be distinguished on the basis of differences in fibre size, lipid content and succinic dehydrogenase and myofibrillar adenosine triphosphatase activity (ATPase activity). Early histological studies have concluded that in addition to a superficial strip of muscle small diameter 'red' fibres occur throughout the trout myotome. These small diameter fibres occurring in the bulk of the myotome are shown to differ from the superficial fibres with respect to their oxidative metabolism and myofibrillar ATPase. The relevance of this finding to studies on the energetics and swimming efficiency of this species are discussed.     

Evolutionary Patterns of Axial Muscle Systems in Some Invertebrates and Fish

SYNOPSIS. Axial muscles used for oscillatory swimming are foundnot only in fish and other vertebrates but also in some protochordatesand invertebrates. Chaetognaths have unsegmented locomotor musculaturewith some unusual features, but larvacean tunicates and thetadpole larvae of ascidians show the simplest variant of thechordate segmented axial muscle arrangement for flexing a notochordalcolumn, where all muscle cells along one side are electricallycoupled. With amphioxus, the basic fish myotomal layout is established,with two main fibre types probably used for different patternsof swimming (as in fish). There are, however, several uniquefeatures, including the flattened fibre shape and the paramyosinsystem of the notochord. Agnatha have two fibre types in themyotomes, a third type perhaps being a developmental stage inthe ontogeny of fast fibres. In lampreys, the central fibresof the characteristic fibre sandwiches in the myotomes are flattened(though less so than in amphioxus); they have a dual innervationof unknown function seen also in the fast fibre system of manyGnathostome fish groups. Hagfish fast fibres are not flattenednor do they have a dual innervation. Gnathostome fish axialmuscles are strikingly uniform in design with two possible exceptions:(1) higher teleost fast fibres which, unlike those of othergroups, are multiply-innervated and (2) tonic fibres in a fewfish, which seem not to be involved in locomotion.     

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.     

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.     

Regional variations in fibre growth dynamics of myotomal and caudal fin muscles in relation to body size of a freshwater teleost, Barbus sarana (Cuv. & Val.)

The growth of red fibres in anterior and middle myotomal regions of B. sarana was mainly by hyperplasia in smaller size classes. In higher size classes, growth by hyperplasia was greater in posterior myotomal region compared to the other two myotomal regions. The growth of pink fibres in anterior myotomal regions was mainly by hypertrophy. The middle and posterior myotomal regions showed fibre growth by hyperplasia. The growth dynamics of white fibres revealed more or less similar pattern in all three myotomal regions against the somatic development. White fibres grew by hyperplasia up to 8 cm F.L. size classes and thereafter by hypertrophy. However, in > 12 cm F.L. size classes, the mean diameter of white fibres did not increase significantly. Similar pattern of growth was found in the white fibres of caudal fin muscle. It is interesting to note that the hyperplasia was mostly completed in the white fibres of the smallest fish studies, whereas, it continued to quite larger fish size in red and pink fibres. Thus, hyperplasia and hypertrophy may be responsible for growth in all fibre types in all myotomal regions in relation to somatic development in this small and medium growing species.     

Dual motor innervation in the axial musculature of fishes

A systematic survey of motor innervation in the myotomal muscle fibres in several groups of fishes was conducted to observe the extent of dual innervation, a phenomenon found within those myotomal muscle fibres with terminal innervation. It is concluded that the dually innervated myotomal muscle fibre is a primitive vertebrate feature. Furthermore, it is suggested that this particular form of polyneuronal innervation is retained in the course of neural evolution within the homogeneous white myotomal muscle fibre region in these fish groups in order to insure synchrony of firing during sudden, rapid locomotion. The findings are examined in light of current ideas regarding the direct adrenergic innervation in addition to cholinergic innervation of striated muscle fibres.     

A morphometric analysis of the spinal motor pool in relation to its target muscle during growth in the trout, Salmo gairdneri Richardson

The relation between number and size of spinal motoneurons and the dimension of myotomal muscle has been investigated in trout at different stages of embryonic, larval and postlarval development (body length 1–15 cm). Three spinal segments have been analysed (cervical, trunk and caudal) and the following parameters were determined by means of a Micromeasurements Image Analyzer: (a) mean cross-sectional myotomal area; (b) mean soma size of principal (or dorsomedian, DM) and secondary (or ventrolateral, VL) motoneurons; (c) DM and VL motoneuron density per segment. Myotomal muscle and motor pool growth was evaluated by percent increments of a, b and c parameters at each stage. Their relationships were denned by equations of computed regression lines.
The analysis provided evidence that: (1) a continuous exponential growth of mean myotomal area takes place in the three segments, with the same trend and with the lowest values in the caudal segment; (2) DM and VL motoneuron size and density per segment also increase during development, with the least value in the caudal segment, VL parameters being of lesser value than DM; (3) motoneuron pool and its target myotomal muscle parameters bear a linear relationship as defined by equations of computer regression lines; (4) motoneuron number percent increment at early eleutherembryonic stage precedes myotomal area increment which takes place during late eleutherembryonic stage.
It is apparent that spinal motor pool and target myotomal muscle grow at the same rate in the trout during the considered stages. The discussion links this fact with the hypothesis of a neuronal influence on muscle fibre type differentiation.     

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.     

Muscle function and swimming in sharks

The locomotor system in sharks has been investigated for many decades, starting with the earliest kinematic studies by Sir James Gray in the 1930s. Early work on axial muscle anatomy also included sharks, and the first demonstration of the functional significance of red and white muscle fibre types was made on spinal preparations in sharks. Nevertheless, studies on teleosts dominate the literature on fish swimming. The purpose of this article is to review the current knowledge of muscle function and swimming in sharks, by considering their morphological features related to swimming, the anatomy and physiology of the axial musculature, kinematics and muscle dynamics, and special features of warm-bodied lamnids. In addition, new data are presented on muscle activation in fast-starts. Finally, recent developments in tracking technology that provide insights into shark swimming performance in their natural environment are highlighted.     

Muscle fibre types and metabolism in post-larval and adult stages of notothenioid fish

Summary A histochemical study was carried out on muscle fibre types in the myotomes of post-larval and adult stages of seven species of notothenioid fish. There was little interspecific variation in the distribution of muscle fibre types in post-larvae. Slow fibres (diameter range 15–60 m) which stained darkly for succinic dehydrogenase activity (SDHase) formed a superficial layer 1–2 fibres thick around the entire lateral surface of the trunk. In all species a narrow band of very small diameter fibres (diameter range 5–62 m), with only weak staining activity, occurred between the skin and slow fibre layer. These have the characteristics of tonic fibres found in other teleosts. The remainder of the myotome was composed of fast muscle fibres (diameter range 9–75 m), which stain weakly for SDHase, -glycerophosphate dehydrogenase, glycogen and lipid. Slow muscle fibres were only a minor component of the trunk muscles of adult stages of the pelagic species Champsocephalus gunnari and Pseudochaenichthys georgianus, consistent with a reliance on pectoral fin swimming during sustained activity. Of the other species examined only Psilodraco breviceps and Notothenia gibberifrons had more than a few percent of slow muscle in the trunk (20%–30% in posterior myotomes), suggesting a greater involvement of sub-carangiform swimming at cruising speeds. The ultrastructure of slow fibres from the pectoral fin adductor and myotomal muscles of a haemoglobinless (P. georgianus) and red-blooded species (P. breviceps), both active swimmers, were compared. Fibres contained loosely packed, and regularly shaped myofibrils numerous mitochondria, glycogen granules and occasional lipid droplets. Mitochondria occupied >50% of fibre volume in the haemoglobinless species P. georgianus, each myofibril was surrounded by one or more mitochondria with densely packed cristae. No significant differences, however, were found in mean diameter between fibres from red-blooded and haemoglobinless species. The activities of key enzymes of energy metabolism were determined in the slow (pectoral) and fast (myotomal) muscles of N. gibberifrons. In contrast to other demersal Antarctic fish examined, much higher glycolytic activities were found in fast muscle fibres, probably reflecting greater endurance during burst swimming.     

Scaling effects on the neuromuscular system,twitch kinetics and morphometrics of the cod,Gadus morhua

Size related changes in muscle twitch kinetics, morphometrics and innervation have been examined in cod, Gadus morhua. Fish size ranged from 9 cm to 45 cm in total length (L).

Twitch contraction time (time to 90% relaxation), scaled in proportion to L^0.29. Scaling of morphometric parameters was essentially geometric. Mean cross‐sectional area and weight of the myotomal muscle scaled in proportion to L^2.05 and L^3.08 respectively. These results are discussed in the light of alterations in length specific swimming performance and kinematics as fish grow.

During growth, the number of endplates terminating on each fast fibre increased, from around 10 on fibres 2 mm in length (～10 cm fish) to 20 on 10 mm fibres (—40 cm fish). However, mean spacing between endplates increased from around 0.25 mm to 0.50 mm. The functional significance of polyneuronal innervation in teleost fast muscle fibres is discussed.     

Red and white muscle fibre activity in swimming skipjack tuna, Katsuwonus pelamis (L.)

To test the hypothesis that white muscle fibre portions of the myotomes are used at sustainable swimming speeds, skipjack tuna, Katsuwonus pelamis , were forced to swim against various current velocities in a water tunnel while electrical activity of the red and white muscle fibres was simultaneously recorded. Eight fish were tested, five fish graded white muscle fibres into activity at swimming speeds above their minimum hydrostatic equilibrium speed, but well below the estimated maximum sustainable swimming speed of skipjack tuna. Three other fish showed white muscle fibre activity at minimum swimming speeds, a possibly abnormal condition.     

Growth dynamics of myotomal muscle fibres in a carangid, Caranx malabaricus (Cuv. et Val.)

The growth pattern of myotomal red, pink and white muscle and its relation to somatic growth in Caranx malabaricus are described. The growth pattern of red muscle was by an increase in fibre number in early size classes (< 22 cm f.l.) and thereafter mainly by increase in fibre diameter and partly by increase in fibre number. The growth of pink muscle was mainly by an increase in fibre diameter, but in smaller fish an increase in fibre number was also evident. White muscle growth was mainly by an increase in fibre diameter and partly by increase in fibre numbers in fish < 22 cm f.l., but only by an increase in fibre diameter from 22 cm f.l. onwards. Caranx malabaricus is a slow-to-moderate growing species and its fibre growth pattern matches with such somatic growth.     

Growth dynamics of white myotomal muscle fibres in the bluntnose minnow, Pimephales notatus Rafinesque, and comparison with rainbow trout, Salmo gairdneri Richardson

The dynamics of increase of the predominant white muscle fibres of the myotomal bulk in bluntnose minnow, Pimephales notatus , ranging from 2.0 to 9.1 cm f.l. have been analysed by examination of modal progression of fibre diameter frequency classes in fish fed to satiation and growing at different rates at 15, 25 or 30°C. Recruitment of new fibres appeared to contribute little to increase in muscle bulk above 4 cm f.l. , and nothing beyond 6 cm. The dominant means of increase was increase in fibre diameter. The limiting fibre diameter seemed to be 120 μ. These dynamics, which result in an approximately 1: 1 ratio between mean fibre diameter and f.l. , are in contrast to those of the myotomal white muscle of rainbow trout, Salmo gairdneri , in which, regardless of differences in somatic growth produced by temperature, ration size or growth hormone administration, mean fibre diameter does not exceed that in bluntnose minnow until trout exceed 30 cm f.l. In trout there is, moreover, input of new fibres up to approximately 50 cm f.l. , when subsequent growth, as in the minnow, is by means of fibre diameter increase. The bluntnose minnow is a small, slow growing species; the rainbow trout is a large, fast growing species. The discussion links these facts with the observed differences in fibre growth dynamics in relation to a hypothesis of interspecific differences in fish growth capability.     

Elevated incubation temperature improves later-life swimming endurance in juvenile Chinook salmon,Oncorhynchus tshawytscha

The effect of incubation and rearing temperature on muscle development and swimming endurance under a high-intensity swimming test was investigated in juvenile Chinook salmon (Oncorhynchus tshawytscha) in a hatchery experiment. After controlling for the effects of fork length (L_F) and parental identity, times to fatigue of fish were higher when fish were incubated or reared at warmer temperatures. Significant differences among combinations of pre- and post-emergence temperatures conformed to 15–15°C > 15–9°C > 9–9°C > 7–9°C > 7–7°C in 2011 when swimming tests were conducted at 300 accumulated temperature units post-emergence and 15–9°C > (7–9°C = 7–7°C) in 2012 when swimming tests were conducted at an L_F of c. 40 mm. The combination of pre- and post-emergence temperatures also affected the number and size of muscle fibres, with differences among temperature treatments in mean fibre cross-sectional area persisting after controlling for L_F and parental effects. Nonetheless, neither fibre number nor fibre size accounted for significant variation in swimming endurance. Thus, thermal carryover effects on swimming endurance were not mediated by thermal imprinting of muscle structure. This is the first study to test how temperature, body size and muscle structure interact to affect swimming endurance during early development in salmon.     

Early locomotor behaviour and the structure of the nervous system in embryos and larvae of the Australian lungfish, Neoceratodusforsteri‡

This paper describes the development of early locomotor responses to mechanical stimulation in the Australian lungfish Neoceralodus and compares them with structural changes in the spinal cord. Initial movements occur spontaneously prior to innervation of myotomal muscles, and are therefore myogenic. After muscle innervation, embryos only move when stimulated; the first type of response to sharp touch is a unilateral flexion away from the stimulus, then at a later stage the contralateral response is followed by a homolateral flexure which, later still, passes into bursts of swimming. The initial contralateral response occurs when decussating interneurons are detectable but before spinal sensory innervation of the trunk; however, the trigeminal sensory pathway has been established by this time and probably mediates the first mechanoreceptive signals from trunk epidermis. Later, Rohon-Beard cells innervate the trunk skin, and then dorsal root neurons take over the major sensory role. The secondary homolateral response and bursts of swimming are paralleled by the development of several types of spinal interneurons and the ingrowth of Mauthner cell axons.     

The effect of starvation on the ultrastructure of the red and white myotomal muscles of the crucian carp (Carassius carassius)

Summary The effect of 16 weeks total starvation on the ultrastructure of the red and white myotomal muscles of the crucian carp (Carassius Carassius) has been investigated. In the white fibres the amount of myofibrillar material fell from 89.6% to 70.7% of the total fibre volume whilst in the red fibres the fall was from 72.2% to 70.3%. The sarcoplasmic reticulum appeared to have become swollen during starvation in both fibre types. In the white fibres the terminal cisternae of some triads seem to have fused. The volume of the red fibres occupied by mitochondria was reduced from 16.2 % to 5.9 %. The concentration of mitochondria in the white fibres was too low to detect any quantitative changes. A marked reduction in the amount of euchromatin material was observed in most white fibre nuclei and many red fibre nuclei. Many of the ultrastructural changes noted in the present study can be correlated with biochemical changes known to occur in the red and white myotomal muscles of fish during starvation. This work was supported by a grant from the Natural Environmental Research Council.     

Resolving Shifting Patterns of Muscle Energy Use in Swimming Fish

Muscle metabolism dominates the energy costs of locomotion. Although in vivo measures of muscle strain, activity and force can indicate mechanical function, similar muscle-level measures of energy use are challenging to obtain. Without this information locomotor systems are essentially a black box in terms of the distribution of metabolic energy. Although in situ measurements of muscle metabolism are not practical in multiple muscles, the rate of blood flow to skeletal muscle tissue can be used as a proxy for aerobic metabolism, allowing the cost of particular muscle functions to be estimated. Axial, undulatory swimming is one of the most common modes of vertebrate locomotion. In fish, segmented myotomal muscles are the primary power source, driving undulations of the body axis that transfer momentum to the water. Multiple fins and the associated fin muscles also contribute to thrust production, and stabilization and control of the swimming trajectory. We have used blood flow tracers in swimming rainbow trout (Oncorhynchus mykiss) to estimate the regional distribution of energy use across the myotomal and fin muscle groups to reveal the functional distribution of metabolic energy use within a swimming animal for the first time. Energy use by the myotomal muscle increased with speed to meet thrust requirements, particularly in posterior myotomes where muscle power outputs are greatest. At low speeds, there was high fin muscle energy use, consistent with active stability control. As speed increased, and fins were adducted, overall fin muscle energy use declined, except in the caudal fin muscles where active fin stiffening is required to maintain power transfer to the wake. The present data were obtained under steady-state conditions which rarely apply in natural, physical environments. This approach also has potential to reveal the mechanical factors that underlie changes in locomotor cost associated with movement through unsteady flow regimes.