Biomedical subjects

I A Johnston

Publications and source records attributed to I A Johnston.

At least 19 recordsLinked to original sources

PubMed2005

The molecular regulation of exercised-induced muscle fibre hypertrophy in the common carp: expression of MyoD, PCNA and components of the calcineurin-signalling pathway.

Growth was investigated over 16 d in juvenile common carp (Cyprinus carpio L.) held in either static water (tank rested, TR16) or exercised in a flume at 2.5-3.2 body lengths s-1 for 18 h a day (exercised, E16). Relative to the start of the experiment (TR0), the TR16 group showed a 31% increase in body mass (specific growth rate, 1.57% d-1), whereas there was no net change in the E16 group. There was, however, a significant exercise-induced hypertrophy of slow muscle fibres with average fibre cross-sectional area (FCSA) increasing by 35% in the E16 group, compared with 11% in the TR16 group. In contrast, FCSA of fast muscle fibres increased by 34% in the TR16 group compared to just 18% in the E16 group. The relative concentrations and subcellular localisation of proteins hypothesised to play a role in the regulation of muscle growth were measured. MyoD concentration was similar in the TR0, TR16 and E16 groups in both slow and fast muscle. However, there was a small (5%-10%) but statistically significant increase in nuclear localisation of MyoD in those groups showing a significant increase in FCSA over the time course of the experiment. PCNA concentration was 31% and 12% higher in the TR16 than in either the TR0 or E16 groups for slow and fast muscle, respectively. Exercise resulted in a approximately 10% increase in nuclear factor of T-cells (NFAT2) concentration in slow muscle but no change in NFAT2 localisation. Calcineurin B concentration was similar in tank rested and exercised groups. The results do not support a major role for the calcineurin-signalling pathway in the regulation of muscle hypertrophy in the common carp.

Animals↗

PubMed2005

The role of myostatin and the calcineurin-signalling pathway in regulating muscle mass in response to exercise training in the rainbow trout Oncorhynchus mykiss Walbaum.

Rainbow trout Oncorhynchus mykiss Walbaum were exercised at 0.8 and 1.6 body lengths s(-1) for 18 h a day over a 30 day 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.

Animals↗

PubMed2002

Peripheral oxygen transport in skeletal muscle of Antarctic and sub-Antarctic notothenioid fish.

Transcellular oxygen flux was modelled mathematically in the aerobic skeletal muscles of perciform fish species living at widely different temperatures (Antarctica, sub-Antarctica and the Mediterranean Sea). Using structural data derived from stereological analysis of electron micrographs, mean fibre P(O(2)) was calculated on the basis of temperature-corrected rates of mitochondrial respiration and oxygen diffusion. The mean muscle fibre diameter (MFD) among Antarctic notothenioids was in the range 17-61 microm and mitochondrial volume density, Vv(mit,f), was 0.27-0.53, but capillary-to-fibre ratio varied only between 1.2 and 1.5. For a mean capillary P(O(2)) of 6 kPa, the model predicted a mean tissue P(O(2)) in the range 0.7-5.8 kPa at the estimated maximum aerobic capacity (M(O(2)max)). The lowest levels of tissue oxygenation were found in the pectoral muscle fibres of the icefish Chaenocephalus aceratus, which lacks the respiratory pigments haemoglobin and myoglobin. Red-blooded notothenioids found in the sub-Antarctic had a similar muscle fine structure to those caught south of the Antarctic Convergence, with an MFD of 20-41 microm and Vv(mit,f) of 0.27-0.33, resulting in an estimated mean P(O(2)) of 4-5 kPa at M(O(2)max). Mean tissue P(O(2)) in the sub-Antarctic icefish Champsocephalus esox, with greater MFD and Vv(mit,f), 56 microm and 0.51, respectively, was calculated to exceed 1 kPa at winter temperatures (4 degrees C), although oxidative metabolism was predicted to be impaired at the summer maximum of 10 degrees C. At the high end of the thermal range, related perciform species from the Mediterranean had a negligible drop in intracellular P(O(2)) across their small-diameter fibres, to a minimum of 5.4 kPa, comparable with that predicted for Trematomus newnesi from the Antarctic (5.6 kPa) with a similar MFD. These data suggest that, within a single phylogenetic group, integrative structural adaptations potentially enable a similar degree of tissue oxygenation over a 20 degrees C range of environmental temperature in the red-blooded notothenioids, and that this is compromised by the lack of respiratory pigments in the icefishes. The mean capillary radius was 1.5 times greater in the two icefish than in the other notothenioids, and the model simulations indicate that the evolution of wide-bore capillaries is essential to maintain tissue oxygenation in the absence of respiratory pigments.

Animals↗

PubMed2001

Plasticity of myosin heavy chain expression with temperature acclimation is gradually acquired during ontogeny in the common carp (Cyprinus carpio L.).

Common carp (Cyprinus carpio L.) were reared from hatching until 61 mm total length (TL) at 21 degrees C. At 14 weeks and 20 weeks post-hatch, corresponding to initial lengths of 30 mm and 44 mm respectively, fish were acclimated to 10 degrees C using a rate of cooling of 1 degrees C per day. A statistical model was used to compare the time course in the change of white muscle myofibrillar ATPase activity with temperature acclimation. The myosin heavy chain (MHC) composition of white muscle myofibrils was investigated using peptide mapping. A significant increase in myofibrillar ATPase activity was observed after 2-3 weeks in the 44 mm group, but not until 4-5 weeks in the 30 mm group. when they had reached 37 mm TL. The MHC banding pattern of 120 mm TL fish acclimated to 10 degrees C or 21 degrees C for a minimum of 6 weeks were distinct from each other. The MHC peptide map characteristic of 10-degrees C-acclimated fish was not observed in individuals less than 37 mm length. We therefore conclude that the capacity to alter the composition and properties of myofibrils with cold acclimation is acquired in juvenile carp at around 37 mm TL.

Acclimatization↗

PubMed2001

Embryonic temperature and the relative timing of muscle-specific genes during development in herring (Clupea harengus L.).

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.

Amino Acid Sequence↗

PubMed2000

Thermal acclimation of locomotor performance in tadpoles and adults of the aquatic frog Xenopus laevis.

Among amphibians, the ability to compensate for the effects of temperature on the locomotor system by thermal acclimation has only been reported in larvae of a single species of anuran. All other analyses have examined predominantly terrestrial adult life stages of amphibians and found no evidence of thermal acclimatory capacity. We examined the ability of both tadpoles and adults of the fully aquatic amphibian Xenopus laevis to acclimate their locomotor system to different temperatures. Tadpoles were acclimated to either 12 degrees C or 30 degrees C for 4 weeks and their burst swimming performance was assessed at four temperatures between 5 degrees C and 30 degrees C. Adult X. laevis were acclimated to either 10 degrees C or 25 degrees C for 6 weeks and their burst swimming performance and isolated muscle performance was determined at six temperatures between 5 degrees C and 30 degrees C. Maximum swimming performance of cold-acclimated X. laevis tadpoles was greater at cool temperatures and lower at the highest temperature in comparison with the warm-acclimated animals. At the test temperature of 12 degrees C, maximum swimming velocity of tadpoles acclimated to 12 degrees C was 38% higher than the 30 degrees C-acclimation group, while at 30 degrees C, maximum swimming velocity of the 30 degrees C-acclimation group was 41% faster than the 12 degrees C-acclimation group. Maximum swimming performance of adult X. laevis acclimated to 10 degrees C was also higher at the lower temperatures than the 25 degrees C acclimated animals, but there was no difference between the treatment groups at higher temperatures. When tested at 10 degrees C, maximum swimming velocity of the 10 degrees C-acclimation group was 67% faster than the 25 degrees C group. Isolated gastrocnemius muscle fibres from adult X. laevis acclimated to 10 degrees C produced higher relative tetanic tensions and decreased relaxation times at 10 degrees C in comparison with animals acclimated to 25 degrees C. This is only the second species of amphibian, and the first adult life stage, reported to have the capacity to thermally acclimate locomotor performance.

Acclimatization↗

PubMed2000

The biomechanics and evolutionary significance of thermal acclimation in the common carp Cyprinus carpio.

The effects of thermal acclimation were investigated in the common carp Cyprinus carpio L. Acclimation and acute temperature effects were tested during ontogeny from larval [9.5 mm total length (L)] to juvenile (69.0 mm L) stages and between 8 and 21 degrees C. The myosin heavy chain (MHC) composition, myofibrillar Mg(2+)-Ca(2+)-ATPase activity, and muscle strains showed significant thermal acclimation effects. MHCs were only expressed in an acclimation temperature-dependent fashion in fish longer than 37 mm. During fast starts, the temperature had a significant effect on the white muscle strain (33% increase and 50% decrease with increasing acclimation and acute temperature, respectively) and contraction duration (25% decrease with increasing acute temperature). Increases in hydrodynamic efficiency (0.19 to 0.38) and hydrodynamic power requirements (Q(10) = 3.2) occurred with increasing acute temperature (10 to 20 degrees C). Competing hypotheses about the evolutionary significance of the temperature acclimation response were tested. Acclimation extended the temperature range for fast-start behavior, but no improvements in performance at the whole animal level were found between 8 and 21 degrees C.

Acclimatization↗

PubMed2000

Phenotypic plasticity of early myogenesis and satellite cell numbers in atlantic salmon spawning in upland and lowland tributaries of a river system.

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.

Adaptation, Physiological↗

PubMed2000

Early thermal experience has different effects on growth and muscle fibre recruitment in spring- and autumn-running Atlantic salmon populations.

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.

Animals↗

PubMed1999

An integrative study of the temperature dependence of whole animal and muscle performance during jumping and swimming in the frog Rana temporaria.

The aims of this study were: (1) to analyze individual variation in frog locomotor performance, (2) to compare the thermal sensitivity of jumping and swimming, and (3) to contrast whole animal versus muscle fiber performance at different temperatures. The jumping and swimming performance of Rana temporaria was analyzed at 5, 10, 15 and 20 degrees C. Muscle fiber bundles were isolated from lateral gastrocnemius and subjected to the length and activation patterns thought to occur in vivo. As temperature increased, locomotor performance in R. temporaria improved with a Q10 of 1.2 for both jump take-off velocity and mean swimming velocity. The slope of the relationship between performance and temperature (TE) was similar for both locomotor parameters and was described by the equation z-scores of locomotor performance = 0.127 x TE - 1.585. Although some frogs performed better than others relative performance was affected by locomotor type and temperature. Locomotor performance improved with temperature as the power required during take-off and the mean muscle power output increased with Q10 values of 1.7 and 1.6 respectively. The mean muscle power output during take-off was only 34% of the calculated requirements for the whole animal, suggesting the involvement of elastic strain energy storage mechanisms.

Animals↗

PubMed1999

Temperature and neuromuscular development in embryos of the trout (Salmo trutta L.).

Myogenesis and neural development were examined in the myotomes of trout (Salmo trutta L.) embryos reared at 2, 6 and 10 degrees C. The relative timings of myotube and muscle fibre formation were similar, with respect to somite stage, at all three temperatures. Myogenesis was seen to begin medially, adjacent to the notochord, and also in separate zones located near the outer surface of the myotomes, believed to be the sites of formation of future slow muscle fibres. Temperature did not affect the relative timings of most aspects of neural development, including HNK-1-immunoreactivity of myosepta, primary motor neuron axonogenesis, Rohon-Beard dendrite outgrowth, and expression of acetylcholinesterase in the spinal chord and at the myosepta. The posterior progression of the lateral line primordium was slightly but significantly delayed relative to somite stage in embryos reared at 10 degrees C compared to 6 and 2 degrees C, while formation of vacuoles in the notochord occurred relatively earlier at higher temperatures. No significant differences in neuromuscular development were observed between offspring of migratory and of non-migratory females.

Acetylcholinesterase↗

PubMed1999

Muscle growth and development in normal-sex-ratio and all-female diploid and triploid Atlantic salmon.

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.

Animals↗

PubMed1998

Muscle power output limits fast-start performance in fish.

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.

Animals↗

PubMed1998

Molecular cloning and developmental expression patterns of the MyoD and MEF2 families of muscle transcription factors in the carp.

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.

Amino Acid Sequence↗

PubMed1997

Temperature and neural development of the Atlantic herring (Clupea harengus L.).

Embryos of Atlantic herring (Clupea harengus L.) from the Buchan (Northern North Sea) stock were incubated from fertilisation until hatching at temperatures of 5, 8, 12, and 15 degrees C. The relative timing of development of the Kolmer-Agduhr (KA) neurons, the posterior lateral line nerve, the motor neurons, and myotubes were determined with respect to somite stage of the embryo. Development of the KA neurons, the lateral line nerve, and the myotubes was similar at all temperatures. In contrast, timing of outgrowth of the motor neuron axons with respect to somite stage was earlier at higher (> or = 12 degrees C) than at lower temperatures (< or = 8 degrees C) although it reached a similar point at all temperatures by the 58-somite stage. Our hypothesis to explain these observations is that delayed motor axon outgrowth in the lower temperature groups is probably due to a delay in a signalling interaction between motor neurons and the somite.

Animals↗

PubMed1995

Isolation of unstable myosins and the analysis of light chains by capillary electrophoresis.

A rapid method for the isolation of unstable fish myosins by Sepharose Q ion-exchange chromatography is described which yields a pure protein essentially free of contamination or breakdown products in less than 6 h. A protocol was developed for determining the molar ratios of myosin light chains (LC) by capillary electrophoresis. The method is quantitative, rapid (<20 min), highly reproducible (<2.1% variation in relative peak migration time), and only uses Femtomole quantities of protein. It was able to separate myosin light chains which could previously only be resolved by 2-D sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Capillary electrophoresis in the presence of SDS gave similar apparent relative molecular masses (M(r)) for most proteins, but an anomalously high M(r) for myosin light chain 3(LC3), as has been reported previously for SDS-PAGE methods.

Animals↗

PubMed1991

Temperature and the energy cost of oscillatory work in teleost fast muscle fibres.

Bundles of 20-30 fast muscle fibres were isolated from the abdominal myotomes of the short-horned sculpin (Myoxocephalus scorpius L.). The energy cost of contraction was measured during oscillatory work at 4 degrees C and 15 degrees C following treatment with iodoacetate and nitrogen gas to block glycolysis and aerobic metabolism. Isolated fibres were subjected to sinusoidal length changes about in situ resting length and stimulated at a selected phase in the strain cycle. Preliminary experiments with untreated preparations established the strain amplitude and stimulation parameters required to maximize work output over a range of cycle frequencies at 4 degrees C and 15 degrees C. Following oscillatory work, treated preparations were rapidly frozen, freeze-dried and the concentrations of phosphocreatine (PCr), creatine, adenosine 5'-triphosphate (ATP), adenosine 5'-di- and mono-phosphate and inosine 5-monophosphate measured by high performance liquid chromatography. The concentration of PCr declined in proportion to the total work done for up to 64 cycles without a significant change in ATP. Maximum power output was produced at a cycle frequency of 5 Hz at 4 degrees C (14-18 W/kg) and 17 Hz at 15 degrees C (23-27 W/kg). The rate of utilization of PCr per cycle was independent of temperature. However, since work per cycle was higher at 4 degrees C (2.7-3.7 mJ/g wet weight) than 15 degrees C (1.2-1.6 mJ/g wet weight), the energetic cost of contraction decreased with increasing temperature.

Adenosine Triphosphate↗

PubMed1991

Inter- and intra-specific variation in myosin light chain and troponin I composition in fast muscle fibres from two species of fish (genus Oreochromis) which have different temperature-dependent contractile properties.

The contractile properties and myofibrillar protein composition of fast muscle have been characterized in pure strains of two tropical fish Oreochromis niloticus and O. andersoni. Single fast muscle fibres were isolated from the abdominal myotomes and chemically skinned. The maximum tension-temperature relationships of fibres were similar at 25-30 degrees C, but diverged below 17 degrees C. At 10 degrees C, maximum tension was around 60% higher in O. andersoni (160 +/- 15 kN m-2) than O. niloticus (105 +/- 13 kN m-2) (mean +/- SD). The myofibrillar protein composition of fast fibres was investigated using one-dimensional and two-dimensional gel electrophoresis and peptide mapping. The two Oreochromis species differed with respect to the composition of myosin light chains, troponin I and myosin heavy chains (V8 protease and chymotrypsin peptide maps). An unexpected finding was the presence of two isoforms of myosin light chain 1 in O. andersoni, with apparent molecular masses of 27.5 kDa (LC1f1) and 26.9 kDa (LC1f2). Individuals with LC1f1 (n = 20) and LC1f1 + LC1f2 (n = 12) were represented in the population studied. The myosin light chain 3 (LC3f) content of fibres was similar in both cases. Breeding experiments established that these intra-specific variations in isoform composition were heritable. Fast muscle from O. niloticus and O. andersoni contain two isoforms of troponin I (TNIfl + TNIf2) which were both expressed in single fibres. The identity of TNI was confirmed using a stationary phase troponin-C affinity column. Of the 20 O. niloticus studied seven contained only TNIf1.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗