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Rainbow trout Oncorhynchus mykiss Walbaum were exercised at 0.8 and 1.6 body lengths s-1 for 18 h a day over a 30day period. Exercise resulted in a 24–30% increase in the average cross-sectional area of fast muscle fibres relative to tank-rested controls. The concentrations of growth factors and transcription factors hypothesised to play a role in regulating exercise-induced muscle fibre hypertrophy were measured. Exercise training resulted in a minor increase in calcineurin localisation in the nucleus. However, nuclear factor of T-cells 2 (NFAT2) nuclear localisation did not follow a pattern that was consistent with NFAT2-mediated transcriptional activity and changes in calcineurin signaling. The active peptide of myostatin, a negative regulator of muscle growth in mammals, was downregulated in exercise groups relative to tank-rested controls, but only by 6–7%. It was concluded that myostatin and calcineurin signaling do not play a major role in regulating exercise-induced muscle hypertrophy in trout. [ABSTRACT FROM AUTHOR]

Temperature influences many aspects of muscle development in herring (Clupea harengus). In Clyde herring, myofibril synthesis occurred later with respect to somite stage in embryos reared at 5 degrees C compared with 12 degrees C. The aim of the present study was to test the hypothesis that the relative timing of expression of myogenic regulatory factors (MRFs) and myosin heavy chain (MyHC) transcripts changes with developmental temperature. Reverse transcriptase/polymerase chain reaction (RT-PCR) was used to clone partial coding regions of MyoD, myogenin and MyHC from juvenile Clyde herring. Embryos were reared at 5, 8 and 12 degrees C, and the spatial and temporal expression patterns of transcripts were investigated using cRNA probes and in situ hybridisation. Antisense probes revealed a rostral-caudal progression of all three transcripts. MyoD transcription initially took place in the adaxial cells of the unsegmented, presomitic mesoderm, whereas myogenin transcription first occurred in newly formed somites. The MyHC gene transcript was not detected until approximately nine somites had formed. Since the somite stage at which the MRFs and MyHC were first expressed was independent of temperature, the hypothesis was rejected. We suggest that the effects of temperature on myofibril synthesis must occur downstream from MyHC transcription either at the level of translation or at the assembly stage.

The consequence of early thermal experience for subsequent growth patterns was investigated in Atlantic salmon (Salmo salar L.). Spring- and autumn-running salmon were caught in upland (Baddoch) and lowland (Sheeoch) tributaries of the River Dee, Aberdeenshire, Scotland, respectively, on the final stages of their spawning migrations. The eggs were incubated at the simulated natural temperature regime of each stream, which was on average 2.8 degrees C lower for the Baddoch. The offspring, representing 11 families per population, were transferred at first feeding to constant environmental conditions (12-14 degrees C; 16h:8h light:dark photoperiod) and reared in replicate tanks. Salmon of both populations were longer and heavier at 6 and 12 weeks in fish initially reared under the cooler Baddoch regime. Length frequency distributions became bimodal after 18 weeks, and only the upper growth mode was studied. Modelling of length distributions at 40 weeks revealed significantly different patterns of muscle growth according to initial temperature regime, but only for the Sheeoch salmon. In fish of Sheeoch origin, significantly more white muscle fibres were recruited per mm(2) increase in myotomal cross-sectional area at Sheeoch than at Baddoch temperatures (P<0.01). After 40 weeks, the density of white fibres was 10.4 % higher in fish initially reared at the Sheeoch (533+/-6 mm(-2)) than at the Baddoch (483+/-5 mm(-2)) thermal regimes (means +/- s.e.m., 16 fish per group; P<0.001). Muscle satellite cells were identified using an antibody to c-met. At 24 weeks, the density of muscle satellite cells was 29 % higher in Sheeoch salmon reared to first feeding at the temperature of their natal stream than at cooler Baddoch temperatures (P<0.01). In contrast, the number and size distributions of white muscle fibres in the myotomes of Baddoch salmon were independent of early thermal experience.

Early myogenesis was studied in the offspring of Atlantic salmon (Salmo salar L.) spawning in a lowland (Sheeoch) and an upland (Baddoch) tributary of the River Dee System, Aberdeenshire, Scotland. Eggs from each population were incubated at the simulated natural thermal regimes of each stream, which was on average 2.8 degrees C cooler for the Baddoch than for the Sheeoch. Relationships between muscle cellularity variables, the density of myonuclei and responses to temperature were investigated using multivariate statistical techniques. These revealed highly significant temperature effects (P<0.001) at hatch (H) and first feeding (FF) and significant interactions between population and temperature (P<0.001), indicating that Baddoch and Sheeoch salmon responded differently to the two temperature regimes. The total cross-sectional area of white muscle (WF.ta) at the adipose fin was relatively independent of temperature at hatch and first feeding in the Sheeoch population. In contrast, for alevins of Baddoch origin, WF.ta was 18.9% (H) and 30.5% (FF) higher in fish incubated at Baddoch than at Sheeoch temperatures. At hatch, there were 15.6% more white muscle fibres (WF.no) at the cooler incubation temperature in fish of Sheeoch origin and 6.0% more in fish of Baddoch origin. However, by first feeding, the difference in WF.no between temperatures had narrowed to 7.2% in the Sheeoch fish and increased to 17.4% in the Baddoch population. In contrast, at hatch, the density of myonuclei was 59.8% higher at the warmer incubation temperature in the Sheeoch population and 23.5% higher in the Baddoch population, but differences were less evident at first feeding. In Baddoch fish, 22.5% of the total muscle nuclei were actively dividing at first feeding, as assessed by staining for proliferating cell nuclear antigen (PCNA). Of the PCNA-positive nuclei, 78% were present in cells that stained for the c-met tyrosine kinase receptor, a marker of satellite cells and their division products. The proportion of c-met-positive cells staining for individual myogenic regulatory factors was 72.4% for the myogenic transcription factor MyoD, 76.3% for the myogenic transcription factor Myf-5, 62.1% for myogenin and 48.7% for the myogenic transcription factor Myf-6. For the Sheeoch population, there were 26.5% more c-met-expressing (P<0.01) and 23.2% more myogenic-regulatory-factor-expressing (P<0.05) cells at Sheeoch than at Baddoch temperatures. In contrast, incubation temperature had no significant effects on satellite cell density in the Baddoch population.

Muscle development and growth were investigated in diploid populations of normal-sex-ratio and all-female Atlantic salmon (Salmo salar L.) and their triploid counterparts produced by high-pressure treatment. Somites were formed at the rate of 6 h-1 in both diploids and triploids at 6 degrees C. The rostral-to-caudal development of myotubes, myofibrils and acetylcholinesterase staining at the myosepta was slightly more advanced in triploid than in diploid fish, although the differences were smaller than among individual families. The c-met receptor tyrosine kinase was used as a molecular marker for the satellite cells involved in postembryonic muscle growth. Satellite cell nuclei comprised 17.5 % of total myonuclei in smolts and they were 24 % more abundant in diploid than in triploid fish. Cells expressing the myogenic regulatory factor myf-6, a marker of satellite cells committed to differentiation, represented 14.8 % of total myonuclei in diploids and 12.5 % in triploids. At ambient temperatures, the number of white muscle fibres in normal-sex-ratio fish increased more than 30-fold between the alevin and smolt stages, and approximately 3.5-fold further during the first year of seawater growth. The rate of muscle fibre recruitment in seawater stages was significantly greater in diploid than in triploid fish, reaching 1162 fibres day-1 and 608 fibres day-1, respectively, in all-female groups 800 days post-hatching. For 42 cm fork-length fish, there were approximately one-third more muscle fibres per myotome in diploid than in triploid groups, 649 878 and 413 619, respectively, for all-female fish. The probability density function of muscle fibre diameters in each fish was estimated using non-parametric smoothing techniques, and the mean densities for diploids (fD) and triploids (fT) were calculated. The peak fibre diameter was approximately 20 (micro)m in all age classes, irrespective of ploidy. Distinct bimodal distributions of muscle fibre diameter were evident in all groups 775 days and 839 days post-hatching, reflecting seasonal cycles of fibre recruitment. fD and fT were compared using a non-parametric bootstrap technique and the reference band representing the null-hypothesis indicated that there was no difference with ploidy. Reference bands for normal-sex-ratio fish at 315 days and 470 days indicated that diploids had a higher percentage of smaller-diameter fibres and that triploid distributions had a thicker right-hand tail. Similar differences in fD and fT of muscle fibre diameters were found for all-female fish, although the statistical evidence was less strong. Reference bands indicated differences in the middle range of the distributions of muscle fibre diameter in fish 620-775 days post-hatch, with triploids having a thicker right-hand tail. Thus, a lower density of satellite cells was associated with reduced rates of fibre recruitment but a compensatory increase in muscle fibre hypertrophy in triploid compared with diploid fish.

cDNA clones encoding the myogenic regulatory factors (MRFs) myogenin, MyoD and myf-5 were isolated by reverse-transcription polymerase chain reaction from larvae and embryos of the common carp (Cyprinus carpio L.). Myocyte-specific enhancer factor 2 (MEF2) cDNAs were identified from a cDNA library from adult carp. Northern blot analysis showed that MyoD, myf-5 and MEF2C transcripts were present in three-somite embryos, whereas myogenin and MEF2A transcripts were not detected until the 15-somite stage. Intense signals of myogenin and MyoD transcripts were observed even in 1-month-old juveniles. Levels of MyoD, myogenin and MEF2A transcripts declined between 1 and 7 months after hatching, and myf-5 gave only a weak signal in the oldest fish. In contrast, levels of MEF2C transcripts were considerably higher in 7-month-old juveniles than in 1-month-old larvae. mRNAs encoding carp myosin heavy chain and alpha-actin were first detected at approximately the time of the first heartbeat, and levels were maximal in juveniles 1 month post-hatching. The relatively high levels of MRF mRNA in juvenile fish probably reflect the recruitment of new muscle fibres from the satellite cell population. It was concluded that the relative importance of the different members of the MyoD and MEF2 families of transcription factors for muscle differentiation changes during ontogeny in the carp.

Fast-starts associated with escape responses were filmed at the median habitat temperatures of six teleost fish: Notothenia coriiceps and Notothenia rossii (Antarctica), Myoxocephalus scorpius (North Sea), Scorpaena notata and Serranus cabrilla (Mediterranean) and Paracirrhites forsteri (Indo-West-Pacific Ocean). Methods are presented for estimating the spine positions for silhouettes of swimming fish. These methods were used to validate techniques for calculating kinematics and muscle dynamics during fast-starts. The starts from all species show common patterns, with waves of body curvature travelling from head to tail and increasing in amplitude. Cross-validation with sonomicrometry studies allowed gearing ratios between the red and white muscle to be calculated. Gearing ratios must decrease towards the tail with a corresponding change in muscle geometry, resulting in similar white muscle fibre strains in all the myotomes during the start. A work-loop technique was used to measure mean muscle power output at similar strain and shortening durations to those found in vivo. The fast Sc. notata myotomal fibres produced a mean muscle-mass-specific power of 142.7 W kg-1 at 20 degrees C. Velocity, acceleration and hydrodynamic power output increased both with the travelling rate of the wave of body curvature and with the habitat temperature. At all temperatures, the predicted mean muscle-mass-specific power outputs, as calculated from swimming sequences, were similar to the muscle power outputs measured from work-loop experiments.

Escape responses (C-shaped fast-starts) were filmed at 500 frames s-1 in the Antarctic rock cod (Notothenia coriiceps) at 0 &deg;C. The activation and strain patterns of the superficial fast myotomal muscle were measured simultaneously using electromyography and sonomicrometry respectively. In order to bend the body into the initial C-shape, the muscle fibres in the rostral myotomes (at 0.35L, where L is total length) shortened by up to 13 % of their resting length at a maximum velocity of 1.68 fibre lengths s-1. During the contralateral contraction, muscle fibres were stretched (by 5 % and 7 % at 0.35L and 0.65L, respectively) and were activated prior to the end of lengthening, before shortening by up to 12 % of resting fibre length (peak-to-peak strain). Representative strain records were digitised to create cyclical events corresponding to the C-bend and contralateral contraction. Isolated fibres were subjected to the abstracted strain cycles and stimulated at the same point and for the same duration as occurs in vivo. During the early phase of shortening, muscle shortening velocity (V) increased dramatically whilst the load was relatively constant and represented a substantial fraction of the maximum isometric stress. Pre-stretch of active muscle was associated with significant force enhancement. For the contralateral contraction, V exceeded that predicted by the steady-state force&shy;velocity relationship for considerable periods during each tailbeat, contributing to relatively high maximum instantaneous power outputs of up to 290 W kg-1 wet muscle mass. In vitro experiments, involving adjusting strain, cycle duration and stimulation parameters, indicated that in vivo muscle fibres produce close to their maximum power. During escape responses, the maximum velocity and acceleration recorded from the centre of gravity of the fish were 0.71&plusmn;0.03 m s-1 and 17.1&plusmn;1.4 m s-2, respectively (mean &plusmn; s.e.m., N=7 fish). Muscle performance was sufficient to produce maximum velocities and accelerations that were within the lower end of the range reported for temperate-zone fish.

Single fibres were isolated from the fast myotomal muscle of the short-horned sculpin (Myoxocephalus scorpius L.). Fish were acclimated to either 5 &deg;C (10 h:14 h light:dark) or 15 &deg;C (12 h:12 h light:dark) for 1&shy;2 months. Isometric tension (Fmax) and unloaded contraction velocity (Vslack) were determined in maximally activated skinned fibres over the range 0 to 20 &deg;C. Fibres isolated from 5 &deg;C-acclimated and 15 &deg;C-acclimated fish failed to relax completely following activations at 15 &deg;C and 20 &deg;C respectively. In 5 &deg;C-acclimated fish, Fmax increased from 75 kN m-2 at 0 &deg;C to 123 kN m-2 at 10 &deg;C and was not significantly higher at 15 or 20 &deg;C. The relationship between Fmax and temperature was not significantly different for cold- and warm-acclimated fish. Vslack was around 2.8 fibre lengths s-1 in both acclimation groups at 0 &deg;C, but it increased at a significantly faster rate with temperature in 15 &deg;C- than in 5 &deg;C-acclimated fish. At 20 &deg;C, Vmax was significantly higher in 15 &deg;C-acclimated (8.7 fibre lengths s-1) than in 5 &deg;C-acclimated fish (5.3 fibre lengths s-1). In order to investigate the molecular mechanism(s) underlying changes in Vmax, myosin was purified by ion-exchange chromatography. No difference in myosin heavy chain composition could be detected on the basis of peptide mapping with four different proteolytic enzymes. Two-dimensional polyacrylamide gel electrophoresis revealed no myofibrillar protein isoforms unique to either acclimation temperature. However, the ratio of myosin alkali light chain contents (LC3f:LC1f), as determined by capillary electrophoresis, was significantly lower in muscle from 15 &deg;C-acclimated (0.73) than from 5 &deg;C-acclimated fish (1.66). The results suggest that changes in Vmax are achieved via altered expression of myosin light chains independently of myosin heavy chain composition. In support of this hypothesis, the myofibrillar ATPase activity of fast muscle was not altered by temperature acclimation.

The swimming kinematics of prey capture was studied in short-horned sculpin (Myoxocephalus scorpius L.) acclimated for 6&shy;8 weeks to either 5 &deg;C or 15 &deg;C (12 h:12 h light:dark) using 15 &deg;C-acclimated shrimps as prey. Fish acclimated to 5 &deg;C remained interested in feeding following an acute rise in temperature to 15 &deg;C over 12 h. Prey capture was a stereotyped behaviour consisting of stalking and stationary phases, followed by an S-shaped fast-start (stage 1), a propulsive stroke (stage 2) and a glide of variable duration during which the mouth was expanded and protruded to suck in the prey (stage 3). The duration of the preparatory stroke (half tail-beat, stage 1) was significantly shorter at 15 &deg;C (48.8 ms) than at 5 &deg;C (108.3 ms) in the 5 &deg;C-acclimated sculpin (Q10=2.2). For 5 &deg;C-acclimated fish, at 5 &deg;C, the maximum values for acceleration and velocity along the path travelled by the fish were 16.2 m s-2 and 0.8 m s-1 respectively. Both variables were relatively independent of acute temperature change (Q10=1.1&shy;1.2; P>0.1). At 15 &deg;C, the maximum velocity was 33 % higher and the tail-beat duration of the propulsive stroke was 37 % shorter in 15 &deg;C-acclimated than in 5 &deg;C-acclimated fish. Both stride length and tail-beat amplitude were significantly higher (28 and 23 % respectively) in 15 &deg;C- compared with 5 &deg;C-acclimated sculpin at 15 &deg;C. The results demonstrate plasticity in the major kinematic variables of fast-starts following warm acclimation sufficient to increase the percentage of successful attacks during prey capture from 23.2 to 73.4 %.

Live fibre bundles were isolated from the fast myotomal muscle of short-horned sculpin (Myoxocephalus scorpius L.) and isometric contractile properties and the force&shy;velocity (P&shy;V) relationship determined at 5, 10 and 15 &deg;C. Experiments were carried out on winter- and summer-caught sculpins and on individuals acclimated for 6&shy;8 weeks to either 5 or 15 &deg;C (12 h:12 h light:dark). Maximum tetanic tension (P0) in fibres from 15 &deg;C-acclimated fish increased from 125 kN m-2 at 5 &deg;C to 282 kN m-2 at 15 &deg;C (R10=2.3). For 5 &deg;C-acclimated fish, P0 was 139 kN m-2 at 5 &deg;C, but fell to 78 kN m-2 at 15 &deg;C, consistent with a partial failure of excitation&shy;contraction coupling at high temperatures. Peak force at 15 &deg;C was increased 2.2 times following depolarisation with a high-K+ solution, but was unaffected by the addition of caffeine and/or eserine to the Ringer's solution. The results from winter- and summer-caught fish were similar to those from 5 &deg;C- and 15 &deg;C-acclimated sculpins respectively. In 15 &deg;C-acclimated fish, the power output of muscle fibres calculated from the P&shy;V relationship was 55 W kg-1 at 5 &deg;C and 206 W kg-1 at 15 &deg;C. The P&shy;V relationship at 5 &deg;C was significantly less curved in muscle fibres from 5 &deg;C- than from 15 &deg;C-acclimated fish. After normalizing the curves for P0 and Vmax, it was found that the change in curvature was sufficient to produce a 40 % increase in relative power output at 5 &deg;C in cold-acclimated fish. The maximum contraction speed of muscle fibres at 15 &deg;C was 2.4 times higher in 15 &deg;C- than in 5 &deg;C-acclimated fish. It was concluded that acclimation modifies the contractile properties of fast muscle fibres at both low and high temperatures.

Short-horned sculpin (Myoxocephalus scorpius L.) were acclimated for 6&shy;8 weeks to either 5 &deg;C or 15 &deg;C (12 h dark: 12 h light). Fast-starts elicited by prey capture were filmed from above in silhouette using a high-speed video camera (200 frames s-1). Outlines of the body in successive frames were digitised and changes in strain for the dorsal fast muscle calculated from a knowledge of backbone curvature and the geometrical arrangement of fibres. For 15 &deg;C-acclimated fish at 15 &deg;C, muscle strain amplitude (peak-to-peak) during the first tail-beat was approximately 0.16 at 0.32L, 0.19 at 0.52L and 0.15 at 0.77L, where L is the total length of the fish. Fast muscle fibres were isolated and subjected to the strains calculated for the first tail-beat of the fast-start (abstracted cycle). Preparations were electrically stimulated at various times after the initiation of the fast-start using an in vivo value of duty cycle (27 %). Prior to shortening, muscle fibres at 0.52L and 0.77L were subjected to a pre-stretch of 0.055l0 and 0.085l0 respectively (where l0 is resting muscle length). The net work per cycle was calculated from plots of fibre length and tensile stress. For realistic values of stimulus onset, the average power output per abstracted cycle was similar at different points along the body and was in the range 24&shy;31 W kg-1 wet muscle mass. During shortening, the instantaneous power output reached 175&shy;265 W kg-1 wet muscle mass in middle and caudal myotomes. At the most posterior position examined, the muscle fibres produced significant tensile stresses whilst being stretched, resulting in an initially negative power output. The fibres half-way down the trunk produced their maximum power at around the same time that caudal muscle fibres generated significant tensile stress. Fast muscle fibres at 0.37&shy;0.66L produced 76 % of the total work done during the first tail-beat compared with only 14 % for fibres at 0.67&shy;0.86L, largely reflecting differences in muscle mass. The effect of temperature acclimation on muscle power was determined using the strain fluctuations calculated for 0.52L. For 5 &deg;C-acclimated fish, the average power per cycle (&plusmn; s.e.m.; W kg-1 wet muscle mass) was 21.8&plusmn;3.4 at 5 &deg;C, falling to 6.3&plusmn;1.8 at 15 &deg;C. Following acclimation to 15 &deg;C, average power per cycle increased to 23.8&plusmn;2.8 W kg-1 wet muscle mass at 15 &deg;C. The results indicate near-perfect compensation of muscle performance with temperature acclimation.

Thermal tolerance and the respiratory properties of isolated red muscle mitochondria were investigated in Oreochromis alcalicus grahami from the alkaline hot-springs, Lake Magadi, Kenya. Populations of O. a. grahami were resident in pools at 42.8 &deg;C and migrated into water reaching temperatures of 44.8 &deg;C for short periods. The maximum respiration rates of mitochondria with pyruvate as substrate were 217 and 284 natom O mg-1 mitochondrial protein min-1 at 37 &deg;C and 42 &deg;C, respectively (Q10=1.71). Fatty acyl carnitines (chain lengths C8, C12 and C16), malate and glutamate were oxidised at 70&shy;80 % of the rate for pyruvate. In order to assess evolutionary temperature adaptation of maximum mitochondrial oxidative capacities, the rates of pyruvate and palmitoyl carnitine utilisation in red muscle mitochondria were measured from species living at other temperatures: Notothenia coriiceps from Antarctica (-1.5 to +1 &deg;C); summer-caught Myoxocephalus scorpius from the North Sea (10&shy;15 &deg;C); and Oreochromis andersoni from African lakes and rivers (22&shy;30 &deg;C). State 3 respiration rates had Q10 values in the range 1.8&shy;2.7. At the lower lethal temperature of O. andersoni (12.5 &deg;C), isolated mitochondria utilised pyruvate at a similar rate to mitochondria from N. coriiceps at 2.5 &deg;C (30 natom O mg-1 mitochondrial protein min-1). Rates of pyruvate oxidation by mitochondria from M. scorpius and N. coriiceps were similar and were higher at a given temperature than for O. andersoni. At their normal body temperature (-1.2 &deg;C), mitochondria from the Antarctic fish oxidised pyruvate at 5.5 % and palmitoyl-dl-carnitine at 8.8 % of the rates of mitochondria from the hot-spring species at 42 &deg;C. The results indicate only modest evolutionary adjustments in the maximal rates of mitochondrial respiration in fish living at different temperatures.

This essay explores how the properties of striated muscles are matched to the tasks they perform during running, swimming and flying. During exercise the major locomotory muscles undergo alternate cycles of lengthening and shortening. Force development is greatly influenced by the timing of stimulation in relation to the length-change cycle and by the nature of elastic structures connecting the muscle fibres to the skeleton. The storage and recovery of elastic strain energy by the tendons (apodema in insects) results in a considerable saving of metabolic energy. Strain is independent of locomotory frequency, body size and muscle temperature. In contrast, the frequency of cycles, and hence strain rate, generally increases with speed and is inversely proportional to body size. The maximum isometric stress (P0) striated muscles can exert is rather similar. During steady running or hopping in mammals the peak muscle stress is around one-third of P0. Behaviours such as vertical jumping impose higher stresses requiring disproportionately larger muscles and tendons, which may limit the storage of elastic strain energy. Muscles of small animals consume significantly more energy per gram than do those of large ones. This may be because they need to activate and deactivate their muscles at a higher rate to move at an equivalent speed. When differences in force production are normalised, by multiplying the energy consumed per stride by stride frequency, similar values for the mass-specific cost of locomotion are found in animals with different leg architectures, numbers of legs, skeletal type, body sizes and muscle temperatures.(ABSTRACT TRUNCATED AT 250 WORDS)

Common carp (Cyprinus carpio L.), 1 kg body weight, were acclimated for 1-2 months to water temperatures of either 7-8 degrees C (cold-acclimated group) or 23-24 degrees C (warm-acclimated group). Single fast fibres and small bundles of slow fibres were isolated from the myotomal muscles and chemically skinned. Force-velocity (P-V) characteristics were determined at 7 degrees C and 23 degrees C. The contractile properties of carp muscle fibres are dependent on acclimation temperature. In the warm-acclimated group maximum isometric tensions (P0, kN m-2) are 47 +/- 6 and 64 +/- 5 for slow muscle fibres and 76 +/- 10 and 209 +/- 21 for fast muscle fibres at 7 degrees C and 23 degrees C, respectively. Maximum contraction velocities (Vmax, muscle lengths-1), are 0.4 +/- 0.05 and 1.5 +/- 0.1 at 7 degrees C (slow fibres) and 0.6 +/- 0.04 and 1.9 +/- 0.4 at 23 degrees C (fast fibres). All values represent mean +/- S.E. P0 and Vmax at 7 degrees C are around 1.5-2.0 times higher for slow and fast muscle fibres isolated from the cold-acclimated group. Fibres from 7 degrees C-acclimated carp fail to relax completely following maximal activations at 23 degrees C. The resulting Ca-insensitive force component (50-70% P0) is associated with the development of abnormal crossbridge linkages and very slow contraction velocities. Activities of enzymes associated with energy metabolism were determined at a common temperature of 15 degrees C. Marker enzymes of the electron transport system (cytochrome oxidase), citric acid cycle (citrate synthase), fatty acid metabolism (carnitine palmitoyl transferase, beta-hydroxyacyl CoA dehydrogenase) and aerobic glucose utilization (hexokinase) have 30-60% higher activities in slow muscle from cold-acclimated than from warm-acclimated fish. Activities of cytochrome oxidase and citrate synthase in fast muscle are also elevated following acclimation to low temperature. It is concluded that thermal compensation of mechanical power output by carp skeletal muscle is matched by a concomitant increase in the potential to supply aerobically-generated ATP at low temperatures.

1. Oxygen consumption, mitochondrial content and enzyme activities were determined in identified muscle fibre types of the 'haemoglobin-less' icefish Chaenocephalus aceratus Lönnberg. 2. Small bundles (2-12) of fast and slow fibres were isolated from the myotomal and superficial pectoral fin abductor muscles, respectively. At 0 degrees C the time to 50% peak force and the half-relaxation time of isometric twitches were, respectively, 18 +/- 1 and 38 +/- 4 ms for fast and 43 +/- 3 and 119 +/- 21 ms for slow muscle fibres (mean +/- S.E.). 3. Measurements of enzyme activities in homogenates suggest that phosphocreatine hydrolysis and oxidative phosphorylation are the main energy-supplying pathways in fast and slow muscles, respectively. Activities of glycolytic enzymes were relatively modest and showed no consistent differences between fibre types. 4. The relationship between oxygen consumption and mitochondria in slow muscle was also determined for a 'red-blooded' antarctic (Notothenia gibberifrons), a cold-temperate (Myoxocephalus scorpius) and a warm-temperate (Oreochromis niloticus) fish. Volume densities of mitochondria were as follows (mean +/- S.D.): C. aceratus, 0.50 +/- 0.08; N. gibberifrons, 0.30 +/- 0.10; M. scorpius, 0.23 +/- 0.05; and O. niloticus, 0.20 +/- 0.05. ADP-stimulated respiration rates were measured in isolated fibre segments. In spite of their different mitochondrial contents, slow fibres from the two antarctic fish utilized pyruvate and palmitoyl-1-carnitine at similar rates (1.0-1.2 mumol O2g-1 wet mass min-1 at 0 degrees C). This suggests that the high density of mitochondria in icefish muscle is related, in part, to diffusion limitations.

1. The differentiation of myotomal muscles in the brook trout (Salvelinus fontinalis Mitchill) has been investigated using p-phenylene diamine stained semi-thin sections and cytochemical and quantitative determinations of enzyme activities. 2. Evidence is presented that the range of fibre size in white muscle represents stages in growth rather than distinct fibre types. 3. Electromyography shows that both red and white muscles are recruited for sustained swimming. The threshold swimming speed for recruitment of white fibres is around 1.8 body length/s (L/s). 4. White muscle citrate synthetase and cytochorome oxidase activities are 25-35% that of red muscle. Hexokinase, phosphorylase and phosphofructokinase activities are 2, 4 and 2 times higher in white than red muscles. It is considered that the aerobic capacity of white muscle is sufficient to support sustained swimming, and that blood glucose could be an important fuel source. 5. Endurance exercise training has been investigated in fish swimming, continuously, for 21 days at 3 L/s. This training regime restricts spontaneous high-speed swimming activity and resulted in a general decline of white muscle glycolytic enzyme activities. Red fibres underwent hypertrophy relative to non-exercised controls (530 +/- 64 microns2 non-exercised, 901 +/- 63 microns2 trained). Aerobic enzyme activities in red muscle and the fraction of fibre volume occupied by mitochondria (30.2 +/- 0.8%) did not change in response to the training programme, but glycolytic enzyme activities were elevated. 3-OH Acyl CoA dehydrogenase activities increased in both red and white muscles indicating an enhanced capacity for fatty acid catabolism with training. 6. Plasma and muscle lactate levels were not statistically different between tank-rested and trained fish swimming a 3 L/s. 7. Adaptations of fish muscle to endurance training are discussed and compared with results for other vertebrates.