50 years of comparative biochemistry: The legacy of Peter Hochachka.

Peter Hochachka was an early pioneer in the field of comparative biochemistry. He passed away in 2002 after 4 decades of research in the discipline. To celebrate his contributions and to coincide with what would have been his 80th birthday, a group of his former students organized a symposium that ran as a satellite to the 2017 Canadian Society of Zoologists annual meeting in Winnipeg, Manitoba (Canada). This Special Issue of CBP brings together manuscripts from symposium attendees and other authors who recognize the role Peter played in the evolution of the discipline. In this article, the symposium organizers and guest editors look back on his career, celebrating his many contributions to research, acknowledging his role in training of generations of graduate students and post-doctoral fellows in comparative biochemistry and physiology.

Dietary lipid quality and mitochondrial membrane composition in trout: responses of membrane enzymes and oxidative capacities.

To examine whether membrane fatty acid (FA) composition has a greater impact upon specific components of oxidative phosphorylation or on overall properties of muscle mitochondria, rainbow trout (Oncorhynchus mykiss) were fed two diets differing only in FA composition. Diet 1 was enriched in 18:1n-9 and 18:2n-6 while Diet 2 was enriched in 22:6n-3. The FA composition of mitochondrial phospholipids was strongly affected by diet. 22:6n-3 levels were twice as high (49%) in mitochondrial phospholipids of fish fed Diet 2 than in those fed Diet 1. 18:2n-6 content of the phospholipids also followed the diets, whereas 18:1n-9 changed little. All n-6 FA, most notably 22:5n-6, were significantly higher in fish fed Diet 1. Nonetheless, total saturated FA, total monounsaturated FA and total polyunsaturated FA in mitochondrial phospholipids varied little. Despite a marked impact of diet on specific FA levels in mitochondrial phospholipids, only non-phosphorylating (state 4) rates were higher in fish fed Diet 2. Phosphorylating rates (state 3), oxygen consumption due to flux through the electron transport chain complexes as well as the corresponding spectrophotometric activities did not differ with diet. Body mass affected state 4 rates and cytochrome c oxidase and F 0 F 1 ATPase activities while complex I showed a diet-specific effect of body mass. Only the minor FA that were affected by body mass were correlated with functional properties. The regulated incorporation of dietary FA into phospholipids seems to allow fish to maintain critical membrane functions even when the lipid quality of their diets varies considerably, as is likely in their natural environment.

Relationship between metabolism, sex and reproductive tactics in young Atlantic salmon (Salmo salar L.).

Atlantic salmon can differ markedly in their growth and in the timing of reproductive maturation, leading to the dramatic contrast between the large anadromous adults and the diminutive mature male parr. This study examined the growth rates, anatomical and physiological characteristics of parr during the adoption of their discrete life histories to ascertain whether these properties can explain tactic choice. To minimise the impact of habitat differences upon these attributes, salmon were reared in the laboratory until 1.5years of age, when the "decisions" to undergo smoltification or to mature as parr had been taken. At 1.5years, both males and females showed bimodal size-frequency distributions. Neither the population of origin nor the paternal reproductive tactic influenced the "decision" to mature or the growth trajectories. Growth rate (% massday(-1) during their final 10months) and the % male and female offspring in the upper modal group were strongly correlated and varied markedly among families. Mean growth rate per family was negatively correlated with mean metabolic rate per family at emergence. Growth rate decreased as a function of parr size in January and the growth rates of upper modal fish were displaced upwards relative to those of lower modal fish. Most males in the smaller size mode matured, whereas all other fish began smoltification. Mature male parr did not differ from similarly sized female pre-smolt in routine metabolic rate, but these smaller fish had higher metabolic rates than larger male and female pre-smolts. However, mature parr differed markedly from similarly sized females and from larger male and female pre-smolts in possessing higher oxidative and lower glycolytic capacities in muscle. Overall, these data are consistent with the interpretation that growth rates dictate the distribution of parr between upper and lower modal groups. Individuals from faster growing families would be more likely to pass the threshold for smoltification and to accelerate growth, whereas those from slower growing families would remain in the lower mode. The use of metabolic capacities, e.g. metabolic rate, was linked with modal group, whereas muscle oxidative capacity was linked with male maturity. Mean family metabolic rate at emergence was negatively linked with mean growth during the subsequent year, suggesting that metabolic efficiency facilitates growth and eventually smoltification.

Alternative developmental pathways and the propensity to migrate: a case study in the Atlantic salmon.

Migratory behaviour with its associated phenotypic changes is generally viewed as an adaptive strategy because it incurs survival or reproductive advantages to migrants. The development of a migrant phenotype is believed to be controlled by threshold mechanisms, where individuals emigrate only after surpassing a particular body size but delay migration if below. For such a strategy to respond to natural selection, part of the phenotypic variance in the propensity to migrate must be explained by variation in additive genetic effects. Here, we use data gathered in the field and from a common rearing experiment to test for a genetic basis associated with seaward migration in Atlantic salmon (Salmo salar L.). We document a high heritability of the liability trait underlying the propensity to emigrate in juvenile salmon, and significant differences between offspring grouped according to their sires in body-size threshold values above which emigration takes place. The presence of additive genetic variance in both the liability and thresholds makes the onset of migration a process sensitive to selection and may therefore constitute an important explanatory mechanism for the interpopulation differences in the size at seaward migration observed in this species.

Do local adaptation and the reproductive tactic of Atlantic salmon (Salmo salar L.) affect offspring metabolic capacities?

Atlantic salmon (Salmo salar L.) is an iteroparous, anadromous species that exhibits some of the greatest within-population variability in size and age at maturity of all vertebrates. In the conditional reproductive strategy of salmonids, the male reproductive tactic expressed is believed to depend on an individual male's status relative to others in the population and therefore depends on his capacity to attain a physiological threshold, the exact nature of which is unknown. Although the threshold is influenced by local biotic and abiotic conditions, it is likely to be under genetic control. Our study examined whether the early growth, muscle metabolic capacities, routine metabolic rate, and spontaneous swimming of salmon alevins reared in laboratory conditions varied with the population of origin, maternal investment, and the paternal reproductive tactic. Our experimental design allowed us to establish that neither the population of origin nor the paternal reproductive tactic influenced the physiological capacities of alevins. The strong influence of the mother on alevin metabolic capacities suggests that the bioenergetic differences in metabolic capacities, realized metabolic rates, and activity levels that could eventually dictate the reproductive tactic of male offspring may originate in maternal effects.

Dietary fatty acid composition changes mitochondrial phospholipids and oxidative capacities in rainbow trout red muscle.

Dietary conditioning of juvenile trout changed the acyl chain composition of mitochondrial phospholipids and the oxidative capacities of muscle mitochondria. Trout were fed three diets differing only in fatty acid (FA) composition. The highly unsaturated 22:6 n-3 (DHA) accounted for 0.4, 14, and 30% of fatty acids in Diets 1, 2 and 3. After 10 weeks of growth, the dietary groups differed markedly in FA composition of mitochondrial phospholipids, with significant dietary effects for virtually all FA. Mean mitochondrial DHA levels were 19, 40 and 33% in trout fed Diets 1, 2 and 3. Mitochondrial oxidative capacities changed with diet, while mitochondrial concentrations of cytochromes and of the adenylate nucleotide translocase (nmol mg(1) protein) did not. Mitochondria from fish fed Diet 1 had higher non-phosphorylating (state 4) rates at 5 degrees C than those fed other diets. When phosphorylating (state 3) rates differed between dietary groups, rates at 5 and 15 degrees C were higher for fish fed the more unsaturated diets. Stepwise multiple regressions indicated that FA composition could explain much (42-70%) of the variability of state 4 rates, particularly at 5 degrees C. At 15 degrees C, FA composition explained 16-42% of the variability of states 3 and 4 rates. Similar conclusions were obtained for the complete data set (trout fed diets 1, 2 and 3) and for the data from trout achieving similar growth rates (e.g. those fed Diets 1 and 2). Neither general characteristics of membrane FA, such as % saturates, unsaturation index, n-3, n-6 or n-3/n-6 nor levels of abundant unsaturated FA such as DHA or 18:1(n-9 + n-7), were systematically correlated with mitochondrial capacities even though they differed considerably between trout fed the different diets. Relatively minor FA (20:5n-3, 20:0, 18:2n-6, 18:3n-3, 18:0 and 15:0) showed better correlations with mitochondrial oxidative capacities. This supports the concept that acyl chain composition modulates mitochondrial capacities via interactions between membrane proteins and specific FA of particular phospholipid classes in their microenvironment.

Food availability and reproduction affects lipid and fatty acid composition of the brown mussel, Perna perna, raised in suspension culture.

We examined the influence of the reproductive cycle and environmental factors on variations of the condition index (CI), tissue dry mass, shell size, total lipid content, and relative percent of fatty acids in the mussel, Perna perna. Spat or juveniles were reared to commercial size (70 mm) in suspension culture in the Golfo de Cariaco, Venezuela between May and October 2004. The dry mass of soft tissues and shell, a visual assessment of gonadal status and the organism lipid profile were established every fortnight. In parallel, we measured the environmental conditions, following chlorophyll a, salinity, temperature and seston levels. After an initial decrease, the CI rose and remained high until August after which it decreased continuously until October. Total lipid values also decreased initially, after which they showed two periods of rapid recuperation and depletion, the first between May and August and the second between August and October. Similar tendencies were noted in the fatty acids, C18:3n-3, C18:4n-3 and C22:6n-3. Correlation analysis found no significant relationships between environmental parameters and the variations in total lipids. However, significant correlations were noted between fatty acids and specific environmental parameters. In particular, temperature was inversely correlated with C14:0, C16:1n-7, C18:0, C18:1n-9 and 20:5n-3. Chlorophyll a was positively correlated with C14:0, C16:1n-7, C18:1n-7, C18:4n-3 and 20:4n-6. On the other hand, gametogenesis had an effect on C14:0, C16:1n-7, C18:1n-9 and C18:1n-7, while spawned and gonadal regression states had an effect on fatty acid 20:4n-6. Temperature and chlorophyll a levels strongly influenced the proportion of mussels spawning, suggesting that their influence upon lipid composition may be secondary to their impact upon reproduction. Despite the thermal stability of this tropical system, the lipid composition of mussels changed markedly during the study, reflecting the central role of diet and reproductive investment upon lipid composition.

Genomewide identification of genes under directional selection: gene transcription Q(ST) scan in diverging Atlantic salmon subpopulations.

Evolutionary genomics has benefited from methods that allow identifying evolutionarily important genomic regions on a genomewide scale, including genome scans and QTL mapping. Recently, genomewide scanning by means of microarrays has permitted assessing gene transcription differences among species or populations. However, the identification of differentially transcribed genes does not in itself suffice to measure the role of selection in driving evolutionary changes in gene transcription. Here, we propose and apply a "transcriptome scan" approach to investigating the role of selection in shaping differential profiles of gene transcription among populations. We compared the genomewide transcription levels between two Atlantic salmon subpopulations that have been diverging for only six generations. Following assessment of normality and unimodality on a gene-per-gene basis, the additive genetic basis of gene transcription was estimated using the animal model. Gene transcription h(2) estimates were significant for 1044 (16%) of all detected cDNA clones. In an approach analogous to that of genome scans, we used the distribution of the Q(ST) values estimated from intra- and intersubpopulation additive genetic components of the transcription profiles to identify 16 outlier genes (average Q(ST) estimate = 0.11) whose transcription levels are likely to have evolved under the influence of directional selection within six generations only. Overall, this study contributes both empirically and methodologically to the quantitative genetic exploration of gene transcription data.

Does condition of Atlantic cod (Gadus morhua) have a greater impact upon swimming performance at Ucrit or sprint speeds?

To compare the sensitivity of sprint and critical (Ucrit) swimming speeds to the condition of Atlantic cod (Gadus morhua) and to identify the best anatomic, behavioural and biochemical correlates of these types of swimming, we established two groups of cod that were fed or starved for 12 weeks. We evaluated sprint swimming and Ucrit performance as well as the speed at which repeated burst-coast movements began in the Ucrit test before measuring the metabolic capacities of red and white muscle sampled caudally, centrally and rostrally and the anatomic characteristics of the cod. White muscle lactate was measured directly after the Ucrit test. As expected, the twofold difference in Fulton's condition factor (0.5+/-0.04 for starved and 1.0+/-0.1 for fed cod) was accompanied by large differences in the anatomic and biochemical parameters measured. Despite the relative sparing of muscle aerobic capacity during starvation and despite the greater use of oxidative fibres during Ucrit compared with sprint swimming, these types of swimming differed by much the same extent between starved and fed cod. In the Ucrit tests, white muscle lactate levels and lactate accumulation per burst-coast movement were considerably higher in fed than starved cod, suggesting more intensive use of fast muscle fibres in cod in good condition. Multiple regression analysis indicated strong correlations between Ucrit, the speed at which regular burst-coasting began and the activity of pyruvate dehydrogenase (PDH) in red muscle (both caudal and central positions). PDH activity may limit the rate of oxidative ATP production by red muscle. The activity of cytochrome c oxidase in rostral white muscle was the strongest correlate of sprint swimming, suggesting that aerobic preparation of white muscle facilitates rapid contraction. The correlation between Ucrit and sprint swimming was weak, perhaps due to inter-individual differences in sensitivity during sprint tests.

Condition, prolonged swimming performance and muscle metabolic capacities of cod Gadus morhua.

This study evaluated the link between swimming endurance and condition of Atlantic cod Gadus morhua that had been fed or starved during the 16 weeks preceding the tests, and assessed whether muscle metabolic capacities explain such links. The condition factor [(somatic mass x fork length(-3))x100] of starved cod was 0.54+/-0.1 whereas that of fed cod was 0.81+/-0.1. In white and red muscle, we measured four glycolytic enzymes: phosphofructokinase (PFK), pyruvate kinase (PK), creatine kinase (CK) and lactate dehydrogenase (LDH), two mitochondrial enzymes: cytochrome c oxidase (CCO) and citrate synthase (CS), a biosynthetic enzyme, nucleoside diphosphate kinase (NDPK), glycogen and protein levels and water content. Muscle samples were taken at three positions along the length of the fish; starvation affected the metabolic capacities of white muscle more than those of red muscle. The levels of glycolytic enzymes and glycogen changed more in white than red muscle during starvation. Both in fed and starved cod, muscle metabolic capacities varied with position along the fish; starvation reduced this longitudinal variation more in white than red muscle. In white muscle of fed cod, the glycolytic enzyme levels increased from head to tail, while in starved cod this longitudinal variation disappeared. In red muscle mitochondrial enzyme levels were highest in the caudal sample, but fewer differences were found for glycolytic enzymes. Swimming endurance was markedly affected by fish condition, with starved fish swimming only 30% of the time (and distance) of fed fish. This endurance was closely linked with the number of burst-coast movements during the test and the activity of CCO and LDH in white muscle. The number of burst-coast movements was significantly linked with condition factor and PFK activity in caudal red muscle and gill arch mass. Our data indicated that cod use both glycolytic and oxidative capacities to support endurance swimming. Furthermore, swimming endurance is linked with the metabolic capacities of red and white muscle.

Once a fast cod, always a fast cod: maintenance of performance hierarchies despite changing food availability in cod (Gadus morhua).

To examine whether Atlantic cod maintain constant hierarchies of sprint speeds and muscle metabolic capacities under different feeding regimes, the physiological capacities of individual cod were followed through a starvation-feeding-starvation cycle. We examined sprint speeds and maximal enzyme activities in white-muscle biopsies at each period. We measured the glycolytic enzymes, phosphofructokinase (PFK) and lactate dehydrogenase (LDH), the mitochondrial enzyme, cytochrome C oxidase (CCO), and the biosynthetic enzyme, nucleotide diphosphate kinase (NDPK). Sprint speeds were measured in a laser diode/photocell-timed raceway. As expected, the feeding regime had a marked impact on the physiological capacities of cod, but the responses differed for sprint-swimming and muscle metabolic capacities. The different enzyme activities as well the condition index generally decreased during the first starvation, improved with feeding, and fell again during the second starvation. In contrast, sprint performance improved after feeding but did not fall with the second starvation. Although both the enzyme activities and the sprint speeds showed considerable interindividual variation, sprint speeds were not significantly correlated with the enzyme activities. The hierarchy of sprint performance of the cod was maintained, regardless of the preceding feeding regime, whereas those of muscle metabolic capacities were not.

Family origin and the response of threespine stickleback, Gasterosteus aculeatus, to thermal acclimation.

To establish whether family origin affects the response of the threespine stickleback (Gasterosteus aculeatus) to thermal acclimation, we examined the rates of feeding, growth, and food conversion, relative tissue and organ masses and activities of a mitochondrial and a glycolytic enzyme in pectoral and axial muscle of individually housed fish from six families during acclimation to 8 degrees C and 23 degrees C. Feeding rates differed among families but were consistently higher in warm-acclimated than cold-acclimated fish. Growth rates differed among families. In four families growth was greater at 8 degrees C; these families generally had higher conversion efficiencies at 8 degrees C than 23 degrees C. For two families, growth was greater at 23 degrees C than 8 degrees C and conversion efficiencies did not differ between 8 degrees C and 23 degrees C. Relative tissue and organ masses (percent axial muscle, hepatosomatic, gut and kidney indices) differed with gender and among families (hepatosomatic, gut and kidney indices) but little with acclimation status. In all families and in both muscles, activities of the mitochondrial enzyme, citrate synthase (CS), were increased by cold acclimation. Axial muscle levels of the glycolytic enzyme, lactate dehydrogenase (LDH), were not affected by thermal acclimation or family origin, but were strongly correlated with the hepatosomatic index and axial muscle protein content. Pectoral muscle levels of LDH were affected by family origin which also influenced the response to thermal acclimation. Similar patterns were observed for specific activities and total muscle contents of these enzymes. Stickleback family origin influenced rates of feeding and growth and the thermal sensitivity of growth rates but not the compensatory increase in muscle CS levels with cold acclimation. The differing thermal sensitivities of growth could reflect distinct strategies for the timing of juvenile growth.

Thermal acclimation, growth, and burst swimming of threespine stickleback: enzymatic correlates and influence of photoperiod.

Threespine sticklebacks (Gasterosteus aculeatus) that had been reared in the laboratory under natural photoperiods were acclimated to 23 degrees and 8 degrees C in late spring under increasing day lengths and again in late fall under decreasing day lengths. The parents of these fish were from the anadromous Isle Verte population. In the spring, cold- and warm-acclimated fish grew at the same rates and attained similar condition factors (mass L(-3)), although food intake was considerably higher at 23 degrees C. As both groups had similar increases in mass and condition, the higher axial muscle activities of citrate synthase and phosphofructokinase (measured at 20 degrees C) after cold acclimation were likely a direct response to temperature. Multiple regression analysis showed that axial muscle levels of cytochrome C oxidase and citrate synthase were correlated with the burst swimming speeds of the spring sticklebacks, while growth rates were positively correlated with lactate dehydrogenase levels in pectoral and axial muscles and creatine kinase levels in the axial muscle. In the fall, the fish in both acclimation groups grew little, although they fed at similar rates as in the spring experiment. Overall, the sticklebacks showed lower burst swimming speeds in the fall. In both spring and fall, the burst speeds of cold- and warm-acclimated sticklebacks only differed at warm temperatures. In the spring experiment, the cold-acclimated fish swam faster, whereas in the fall experiment the warm-acclimated fish swam faster despite their lower percentage of axial muscle. Swimming speeds were measured both at a fish's acclimation temperature and after 12 h at the other temperature. Cold-acclimated sticklebacks seem to have more facility in rapidly adjusting to warm temperatures when they have experienced increasing rather than decreasing day lengths, perhaps as a result of the requirements of the spring migration to the intertidal breeding grounds.

Biochemical indicators of muscle growth in the snow crab Chionoecetes opilio (O. Fabricius).

This study examined the relationships between muscle growth rate, the activity of metabolic enzymes and the RNA:DNA ratio, in adult snow crabs Chionoecetes opilio. After moulting, crabs were assigned to three feeding rations to attain a range of tissue growth rates. Muscle growth rate, estimated by the variation in dry tissue content per ml of merus of the first walking leg, was positively correlated with changes in muscle cell number, as evaluated by the DNA content per ml of merus. However, no significant correlation was detected between growth rate and the variation in muscle cell size, the latter being estimated by the change in the protein:DNA ratio. This is due to the fact that, in starved crabs, a reduction in the number of cells is partly compensated by a size increment of the remaining ones. This phenomenon also weakened the overall relationship between muscle growth rate and the phosphofructokinase (PFK) capacity per ml of merus. The simple correlation between those two variables was significantly positive for animals which increased their mass of muscle but insignificant for those which were loosing muscle mass. The lactate dehydrogenase (LDH), citrate synthase (CS) and cytochrome c oxidase (CCO) capacity per ml of merus did not match growth rate. The significant simple correlations that were detected between growth rate and the various enzyme activity expressed per g of protein, per µg of DNA and per g of dry mass did not hold when partial correlations were computed. Variations in muscle cell size were related to adjustments in the quantity of RNA per cell, as depicted by the RNA:DNA ratio. Since muscle growth was not correlated with the variation in muscle cell size, it was not correlated with the RNA:DNA ratio either.

Effects of voluntary activity and genetic selection on muscle metabolic capacities in house mice Mus domesticus.

Selective breeding is an important tool in behavioral genetics and evolutionary physiology, but it has rarely been applied to the study of exercise physiology. We are using artificial selection for increased wheel-running behavior to study the correlated evolution of locomotor activity and physiological determinants of exercise capacity in house mice. We studied enzyme activities and their response to voluntary wheel running in mixed hindlimb muscles of mice from generation 14, at which time individuals from selected lines ran more than twice as many revolutions per day as those from control (unselected) lines. Beginning at weaning and for 8 wk, we housed mice from each of four replicate selected lines and four replicate control lines with access to wheels that were free to rotate (wheel-access group) or locked (sedentary group). Among sedentary animals, mice from selected lines did not exhibit a general increase in aerobic capacities: no mitochondrial [except pyruvate dehydrogenase (PDH)] or glycolytic enzyme activity was significantly (P < 0.05) higher than in control mice. Sedentary mice from the selected lines exhibited a trend for higher muscle aerobic capacities, as indicated by higher levels of mitochondrial (cytochrome-c oxidase, carnitine palmitoyltransferase, citrate synthase, and PDH) and glycolytic (hexokinase and phosphofructokinase) enzymes, with concomitant lower anaerobic capacities, as indicated by lactate dehydrogenase (especially in male mice). Consistent with previous studies of endurance training in rats via voluntary wheel running or forced treadmill exercise, cytochrome-c oxidase, citrate synthase, and carnitine palmitoyltransferase activity increased in the wheel-access groups for both genders; hexokinase also increased in both genders. Some enzymes showed gender-specific responses: PDH and lactate dehydrogenase increased in wheel-access male but not female mice, and glycogen phosphorylase decreased in female but not in male mice. Two-way analysis of covariance revealed significant interactions between line type and activity group; for several enzymes, activities showed greater changes in mice from selected lines, presumably because such mice ran more revolutions per day and at greater velocities. Thus genetic selection for increased voluntary wheel running did not reduce the capability of muscle aerobic capacity to respond to training.

Longitudinal and allometric variation in indicators of muscle metabolic capacities in atlantic cod (Gadus morrhua).

This study evaluated whether indicators of metabolic capacity of cod white muscle differ along the length of the body, whether this variation persists over a large range of body sizes, and whether the allometry of metabolic capacities is similar along the length of the body. We examined the maximal activities of two glycolytic enzymes, phosphofructokinase (PFK) and lactate dehydrogenase (LDH), a mitochondrial enzyme, cytochrome C oxidase (CCO), and the biosynthetic enzyme nucleotide diphosphate kinase (NDPK). All enzymes examined showed significant size dependence, which was generally apparent in all regions. The activity of glycolytic enzymes increased with size, whereas that of CCO and NDPK decreased with size. For PFK and LDH, the size dependence decreased caudally, whereas for CCO and NDPK it was strongest in the caudal sample. For each size range, the activities of PFK, LDH, and CCO were higher in the last third of the body than in the middle or just behind the head. In contrast, NDPK activity was higher just behind the head than at the middle or in the last third of the body, suggesting that nuclear proliferation is more rapid in this zone. The high capacity for adenosine triphosphate (ATP) generation in the caudal region suggests that increases in mass-specific ATP output are advantageous in this relatively thin section of the body.

Effect of diet and temperature upon muscle metabolic capacities and biochemical composition of gonad and muscle in Argopecten purpuratus Lamarck 1819.

Recently spawned Argopecten purpuratus broodstock were conditioned at two temperatures and fed three different diets (microalgae, microalgae mixed with lipids and microalgae mixed with carbohydrates) to examine changes in the biochemical composition of gonad and muscle as well as muscle metabolic capacities. During one experiment, scallops were fed at 3% of their dry mass per day whereas during a second experiment, they were fed at 6% of their dry mass per day. During both experiments, total gonadal levels of lipids and protein increased markedly during conditioning with the two mixed diets at 16 degrees C. These increases were less pronounced at 20 degrees C. Carbohydrate gonadal levels only increased during the second experiment at both temperatures and with the three diets. Of the major biochemical components of the adductor muscle, carbohydrate levels changed most during conditioning. Whereas muscle protein levels increased slightly with gonadal maturation, carbohydrate levels dropped considerably. Despite the marked drop in the levels of glycolytic substrates, only the activities of octopine dehydrogenase in the adductor muscle of the scallops conditioned at 16 degrees C consistently decreased. Muscle levels of glycogen phosphorylase were higher in mature than in recently spawned (control) scallops, suggesting a role in the transfer of glucose equivalents from the adductor muscle to other tissues.

Seasonal cycles of mitochondrial ADP sensitivity and oxidative capacities in trout oxidative muscle.

Mitochondria from red myotomal muscle of rainbow trout, Oncorhynchus mykiss, showed seasonal cycles of their maximal rates of substrate oxidation (nmol.min-1 mg-1 mitochondrial protein) and their apparent ADP affinity (Kmapp), as well as in the thermal sensitivity of these properties. Increases in the maximal capacity of pyruvate oxidation were sufficient to compensate for seasonal changes in temperature, except during the winter months when rates at habitat temperature were depressed relative to other periods. The ADP affinity of isolated mitochondria was highest during cold months. Thus, the Kmapp for ADP at habitat temperature showed less seasonal variation than the ADP Kmapp at a given temperature. A loss in ADP affinity with decreasing temperature occurred through much of the year, and only was definitively suppressed in December and July. Both the ADP affinity and the maximal oxidative capacities of muscle mitochondria seem to be regulated parameters.

Changes in qualitative composition of white muscle with nutritional status of Atlantic cod, Gadus morhua.

Proteins from white muscle are mobilized to cover energy requirements during long-term starvation in fish. Using SDS-polyacrylamide gel electrophoresis, we compared the soluble and insoluble fractions of white muscle proteins from fed and starved Atlantic cod, Gadus morhua, to establish whether preferential preservation or degradation of specific proteins occurred during starvation. While starvation induced no qualitative changes in the electrophoretic pattern of the myofibrillar fraction, our results document differential decreases in the levels of soluble proteins during fasting. Moreover, immunoblot analysis using a monoclonal antibody directed against actin, showed a marked accumulation of this protein in the sarcoplasmic fraction of starved individuals, most likely due to myofibrillar degradation.