Starvation-induced impairment of metabolism in a freshwater catfish

Starvation induced changes in citrate synthase (CS), glucose-6-phosphate dehydrogenase (G6-PDH), lactate dehydrogenase (LDH), DNA, RNA, RNA/DNA ratio and protein were studied in the freshwater catfish Clarias batrachus. Starvation gradually decreased the activity of CS, G6-PDH and LDH in brain, liver and skeletal muscle of the freshwater catfish. The maximum reduction in these enzyme activities upto 35-45% was observed after 35 days of fasting. This shows substantial decline in aerobic and biosynthetic capacity during starvation period. DNA, RNA, RNA/DNA ratio and protein contents were also reduced from 40-67% which reflects reduction in an overall capacity of the protein synthesis. Starvation-induced macromolecular changes indicate impairment of metabolism in fish.     

Differential effects of thyroxine on metabolic enzymes and other macromolecules in a freshwater teleost

The effects of thyroxine (T(4)) on citrate synthase (CS), glucose 6-phosphate dehydrogenase (G6-PDH), lactate dehydrogenase (LDH), DNA, RNA, and protein of various tissues were studied to elucidate the hormonal control of metabolism in a freshwater catfish, Clarias batrachus. T(4) did not produce any significant effect on DNA content of the fish. The CS, RNA, and protein contents of brain, liver, and skeletal muscle of the fish exposed to thiourea for 28 days decreased approximately 50-58% as compared to their levels in control individuals. Injection of T(4) to thiourea-exposed fish produced about three-fold increases in CS, RNA, and protein. These macromolecular inductions by T(4) were blocked by actinomycin D or cycloheximide. This suggests T(4)-induced de novo synthesis of macromolecules and stimulation of aerobic capacity. However, the activities of G6-PDH and LDH of brain, liver, and skeletal muscle of the fish exposed to thiourea increased two times that of the activities in control individuals. Administration of T(4) to thiourea-exposed fish reduced LDH and G6-PDH activities by about 64-74%, which reflects T(4)-dependent inhibition in anaerobic power and selective anabolic activities of the HMP pathway. These differential effects of T(4) on some metabolic enzymes and other important macromolecules may be to meet the other T(4)-induced responses in the freshwater catfish.     

Sex-specific metabolic changes in the annual reproductive cycle of a freshwater catfish

Sex-specific enzymatic and other biochemical changes were studied in the annual reproductive cycle of the freshwater catfish, Clarias batrachus. Citrate synthase (CS) activity of brain, liver and skeletal muscle was maximum in spawning and minimum in postspawning showing a sharp decline in aerobic capacity after spawning. Similar CS activity in remaining phases of the annual reproductive cycle reflects similar energy need during regressed, preparatory and prespawning phases. Glucose 6-phosphate dehydrogenase (G6-PDH) activity declined in spawning and postspawning indicating a possible decrease in lipid and nucleic acid syntheses. The subsequent increase in G6-PDH activity with onset of resting and maintenance of the increased level throughout preparatory and prespawning phases shows restoration of biosynthetic activity. Higher activity of G6-PDH in female than male may be to satisfy a greater biosynthetic need of female reproduction and breeding. The decreased RNA content of tissues showed reduction in protein synthesis capacity during spawning and subsequent increase through postspawning until resting phase. The RNA content of brain and liver was higher in female than male during preparatory and prespawning, which may be associated with higher protein synthesis requirement of female for preparation of reproductive activities. The requirement based sex related changes in metabolism of catfish may be enzyme, tissue or reproductive phase-specific.     

50 Hz magnetic fields activate mussel immunocyte p38 MAP kinase and induce HSP70 and 90

We studied the effects of a sublethal concentration of pyrethroid insecticide fenvalerate on metabolic enzymes, RNA and protein of brain, liver and skeletal muscle of the freshwater catfish, Clarias batrachus. Exposure to fenvalerate gradually decreased the activity of citrate synthase (CS), glucose 6-phosphate dehydrogenase (G6-PDH) and lactate dehydrogenase (LDH) in brain, liver and skeletal muscle up to 21 days. The maximum decrease in enzyme activity was 23-47%. Withdrawal of fenvalerate from the medium for 21 days restored enzyme activity to their control level in all three tissues. RNA and protein content in brain, liver and skeletal muscle decreased significantly with exposure of fenvalerate up to 21 days. The maximum decrease in RNA and protein was 22-32%. Withdrawal of fenvalerate from the medium for 21 days restored the RNA and protein contents to control levels. The present study suggests that fenvalerate impairs cellular metabolism and its biochemical effects are reversible after withdrawal of fenvalerate.     

Scaling effects on metabolism of a teleost

Scaling effects on citrate synthase (CS), glucose-6-phosphate dehydrogenase (G6-PDH), RNA. RNA/DNA ratio and protein contents of brain, liver and skeletal muscle were studied in a teleost, Clarias batrachus. The activity of white skeletal muscle CS decreased significantly as a function of increasing body mass of the fish. It shows that the fulfilment of energy demand in white skeletal muscle is not dependent on aerobic metabolism. The activity of liver G6-PDH decreased with the increasing body mass showing reduction in NADPH generation for lipogenic activity. However, increase in G6-PDH activity showed enhancement in reductive synthesis in skeletal muscle of the larger-sized individuals. A positive scaling of RNA, RNA/DNA ratio and protein contents reflects changes in macromolecular turnover for ATP-supplying enzymes and proteins.     

砷对兰州鲇组织中代谢酶活性及RNA和蛋白质含量影响

采用毒性试验方法,研究了安全浓度（1.288 mg/L）条件下亚砷酸钠（NaAsO2）对兰州鲇（Silurus lanzhouensis）脑、鳃、肝脏、肌肉4种组织中6-磷酸葡萄糖脱氢酶（G-6-PDH）和乳酸脱氢酶（LDH）活性,以及RNA和蛋白质含量的影响。结果表明,染毒21d时,As（Ⅲ）可显著降低4种组织中G-6-PDH和LDH活性、RNA和蛋白质含量（P0.05）。撤毒后21d,除脑和肝组织中蛋白质含量未恢复到对照组水平（P0.05）,肝脏中G-6-PDH活性超过了对照组水平（P0.05）外,其余各组织中G-6-PDH和LDH活性、RNA和蛋白质含量均可恢复到对照组水平（P0.05）。以上结果表明,As（Ⅲ）对兰州鲇组织中代谢酶活性具有明显的抑制作用,可致组织细胞RNA损伤和可溶性蛋白质减少,但这种影响是可逆的,撤毒后一定时间内可恢复到正常水平。       

Scaling of some metabolic enzymes in liver of a freshwater teleost: an adaptive mechanism

The activities of mitochondrial malate dehydrogenase (mMDH) and the total mitochondrial proteins increase as a function of body mass in the freshwater catfish, Clarias batrachus. It clearly indicates an increase in energy production in larger-sized individuals for various purposes including prey-predator interactions. The higher activity of lactate dehydrogenase (LDH) in larger fish may indicate more production of lactate for gluconeogenesis in the liver to meet emergency requirements of increased energy demand. However, the activity of cytoplasmic malate dehydrogenase (cMDH) decreases with the increasing body mass of the fish which reflects reduction in NADPH production and, in turn, reduced lipogenesis in liver of larger individuals. Thus, the present observations suggest an adaptive mechanism dealing with the higher energy budget, and reduced synthetic activities (lipogenesis) in the liver of larger-sized freshwater catfish. This type of biochemical scaling might be also supporting other metabolic pathways in order to adjust some physiological functions for survival in the aquatic environment.     

Physiological correlates of seasonal growth patterns in lake trout Salvelinus namaycush

Physiological correlates of seasonal growth patterns were measured in lake trout Salvelinus namaycush from two populations with contrasting diets (zooplankton-dominated diet in Louisa Lake; fish-dominated diet in Opeongo Lake). Fish in Opeongo Lake grew faster and were in better condition than fish in Louisa Lake. The most prominent biochemical difference between populations was higher citrate synthase (CS) and cytochrome c oxidase activity in the white muscle of fish from Opeongo Lake, indicating greater sustained swimming activity in this lake. In contrast, lactate dehydrogenase (LDH) activity in white muscle, an indicator of capacity for burst swimming, was similar between lakes. Nucleoside diphosphate kinase (NDPK) activity in white muscle, an indicator of protein synthesis, was higher in Opeongo Lake than in Louisa Lake but only in the autumn. In both lakes, protein concentration and therefore nutritional status increased as the growing season progressed from spring to summer to autumn. Biochemical indicators of growth and activity showed similar seasonal patterns in the two lakes with the spring characterized by high NDPK, high CS and high LDH activities (i.e. high levels of protein synthesis in association with high aerobic and anaerobic activities). These results suggest high foraging effort and allocation to growth early in the growing season in both lakes.     

High swimming and metabolic activity in the deep-sea eel Synaphobranchus kaupii revealed by integrated in situ and in vitro measurements

Several complementary studies were undertaken on a single species of deep-sea fish (the eel Synaphobranchus kaupii) within a small temporal and spatial range. In situ experiments on swimming and foraging behaviour, muscle performance, and metabolic rate were performed in the Porcupine Seabight, northeast Atlantic, alongside measurements of temperature and current regime. Deep-water trawling was used to collect eels for studies of animal distribution and for anatomical and biochemical analyses, including white muscle citrate synthase (CS), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), and pyruvate kinase (PK) activities. Synaphobranchus kaupii demonstrated whole-animal swimming speeds similar to those of other active deep-sea fish such as Antimora rostrata. Metabolic rates were an order of magnitude higher (31.6 mL kg(-1) h(-1)) than those recorded in other deep-sea scavenging fish. Activities of CS, LDH, MDH, and PK were higher than expected, and all scaled negatively with body mass, indicating a general decrease in muscle energy supply with fish growth. Despite this apparent constraint, observed in situ burst or routine swimming performances scaled in a similar fashion to other studied species. The higher-than-expected metabolic rates and activity levels, and the unusual scaling relationships of both aerobic and anaerobic metabolism enzymes in white muscle, probably reflect the changes in habitat and feeding ecology experienced during ontogeny in this bathyal species.     

Changes in the metabolic profile of the equine gluteus medius as a function of sampling depth

1. Cross sections from the middle of the gluteus medius were removed from 10 adult horses and used to evaluate changes in histochemically determined muscle fiber type and biochemically determined metabolic enzyme activities as a function of sample depth. 2. Muscle fiber types determined using histochemical methods for myosin ATPase (pH 9.4) and succinic dehydrogenase (SDH) activity indicated percent fast-twitch glycolytic (FG) muscle fibers decreased and slow-twitch oxidative (SO) fibers increased as a function of increasing sampling depth. 3. Percent histochemically determined fast-twitch oxidative glycolytic (FOG) fibers decreased slightly only in the deepest region of the gluteus medius. 4. Citrate synthase (CS) enzymatic activity, used as a marker for mitochondrial oxidative potential, increased 2.5-fold in activity per g of muscle protein from 1 to 8 cm sampling depth. 5. 3-hydroxyacyl-CoA dehydrogenase (HAD) enzymatic activity, used as a marker for lipid oxidation potential, increased 3-fold in activity per g of muscle protein when the depth increased from 1 to 8 cm. 6. Phosphorylase (PS) enzymatic activity, used as a marker for potential glycogen utilization, decreased 50% in activity per g of muscle protein when going from 1 to 8 cm. 7. Lactate dehydrogenase (LDH) enzymatic activity, used as a marker for anaerobic glycolytic potential, decreased about 50% in activity as the sampling depth increased from 1 to 8 cm. 8. In summary, the superficial portion of the equine gluteus medius was found to be more glycolytic and less aerobic in its metabolic profile than deeper regions. The muscle became progressively more aerobic and less glycolytic with increasing sampling depth.     

Effect of tuftsin on enzyme activity in the energy metabolism of lymphocytes

The cytochemical technique was used to measure the activity of succinate dehydrogenase (SDH), lactate dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) of peripheral blood lymphocytes of mice and rats given intraperitoneal injections of an endogenous immunostimulant tuftcin (Tre-Lys-Pro-Arg) in a dose of 0.3 mg/kg. A significant decrease of SDH activity was observed both in mice and rats 4 and 6 hours following injection, respectively. In mice, that activity returned to normal in 12, while in rats in 24 hours. An opposite action was produced by tuftcin on G-6-PDH, causing the maximum elevation of the enzyme activity in rat lymphocytes 6 hours after peptide administration. The decrease to the initial level was observed in 24 hours. Tuftcin did not affect the activity of LDH. The data obtained indicate that the immunological effect of tuftcin is coupled with the changes in the activity of Krebs cycle enzymes (SDH) and pentose phosphate cycle enzymes (G-6-PDH).     

Adaptations in metabolic capacity of rat soleus after paralysis.

To determine whether long-term reductions in neuromuscular activity result in alterations in metabolic capacity, the activities of oxidative, i.e., succinate dehydrogenase (SDH) and citrate synthase (CS), and glycolytic, i.e., alpha-glycerophosphate dehydrogenase (GPD), enzyme markers were quantified in rat soleus muscles 1, 3, and 6 mo after a complete spinal cord transection (ST). In addition, the proportional content of lactate dehydrogenase (LDH) isozymes was used as a marker for oxidative and glycolytic capacities. The myosin heavy chain (MHC) isoform content of a fiber served as a marker of phenotype. In general, MHC isoforms shifted from MHC1 toward MHC2, particularly MHC2x, after ST. Mean SDH and CS activities were higher in ST than control at all time points. The elevated SDH and CS activities were indicative of an enhanced oxidative capacity. GPD activities were higher in ST than control rats at all time points. The increase in activity of SDH was larger than GPD. Thus the GPD-to-SDH (glycolytic-to-oxidative) ratio was decreased after ST. Compared with controls, total LDH activity increased transiently, and the LDH isozyme profile shifted from LDH-1 toward LDH-5, indicative of an enhanced glycolytic capacity. Combined, these results indicate that 1) the metabolic capacities of soleus fibers were not compromised, but the interrelationships among oxidative and glycolytic capacity and MHC content were apparently dissociated after ST; 2) enhancements in oxidative and glycolytic enzyme activities are not mutually exclusive; and 3) chronic reductions in skeletal muscle activity do not necessarily result in a reduced oxidative capacity.     

The effect of torbafylline on enzyme activities in fast and slow muscles with limited blood supply.

1. Activities of a glycolytic enzyme--lactate dehydrogenase, LDH, and two oxidative enzymes--citrate synthase (CS), a marker for TCA cycle entry, and 3-hydroxyacyl-CoA dehydrogenase (HAD), which indicates the capacity for beta-oxidation of endogenous lipids, were measured in fast (tibialis anterior, TA, and extensor digitorum longus, EDL) and slow (soleus, SOL) muscles of Sprague-Dawley rats with intact and limited blood supply, and following treatment with the xanthine derivative torbafylline (Hoechst, Werk Albert, Wiesbaden). 2. Limitation of blood supply by unilateral ligation of the common iliac artery increased activity of LDH in fast muscles, and activity of CS and HAD in soleus. 3. Torbafylline treatment caused an increased LDH activity in intact fast muscles and decreased it in soleus, although the relative capacity for anaerobic and aerobic metabolism (indicated by the ratio of LDH and CS activities) remained unchanged in all cases. 4. Whilst having little effect on oxidative enzyme activity of fast muscles, torbafylline decreased the activity of CS but increased activity of HAD in soleus, suggesting a greater reliance on lipid metabolism. 5. The effect of arterial ligation on enzyme activity was ameliorated by treatment with torbafylline, possibly due to its effect on the microcirculation.     

Ontogenetic changes in citrate synthase and lactate dehydrogenase activity in the jumping muscle of the American locust (Schistocerca americana)

Intraspecific studies have repeatedly shown that muscle-specific oxidative enzyme activities scale negatively with body mass while muscle-specific glycolytic enzyme activities scale positively. However, most of these studies have not included juveniles. In this study, we examined how citrate synthase (CS, EC 2.3.3.1) and lactate dehydrogenase (LDH; EC 1.1.1.27) activity in the jumping muscle of Schistocerca americana grasshoppers varied with ontogeny across a 40-fold increase in body size. In contrast to the pattern observed when adult conspecifics are compared, we show that jumping muscle CS activity increased more than 2-fold from 2nd instars to adults, while jumping muscle LDH activity increased more than 5-fold. The increased LDH activity in older grasshoppers supports previous data that older grasshoppers have a reduced jumping endurance. The increased CS activity with age may help older grasshoppers efficiently produce aerobic ATP to bend cuticular springs for energy storage before a jump or alternatively recover from anaerobic metabolism after jumping. Metabolic changes in S. americana jumping muscle are similar to other developing taxa and highlight the importance of including juveniles within intraspecific studies. When compared to adults, juvenile locomotion may have increased selection pressure because of both greater energetic demands during growth and higher predation rates.     

Enzymatic activity in crude oil contaminated rats

The activity of enzymes found in the plasma, malate dehydrogenase (MDH) and lactate dehydrogenase (LDH), and enzymes from erythrocytes, glucose-6-phosphate dehydrogenase (G-6-PDH) and catalase, was studied in rats contaminated by crude oil. Crude oil (tube fed) contamination caused a significant increase in MDH and LDH activity 96 hr after contamination while a decrease in activity was noted in 6-6-PDH and catalase. An additional contamination (1 week after the first contamination), measured 96 hr after contamination, caused a relative decrease in MDH and LDH activity while there was a contrasting relative increase in G-6-PDH and catalase activity. After a recovery period of 3 weeks the only significant change was an increase in catalase activity.     

Aerobic capacity of frog skeletal muscle during hibernation

Frogs submerged at 3 degrees C in hypoxic water (Po2=60 mmHg) depress their metabolic rate to 25% of that seen in control animals with access to air. The hypometabolic state of the skeletal muscle in such cold-submerged frogs is thought to be the most important contributor to the overall metabolic depression. The aim of this study was to determine whether the aerobic capacity of frog skeletal muscle became altered during 1-4 mo of hibernation to match the reduction in adenosine triphosphate (ATP) demand. To this end, the activities of key mitochondrial enzymes were measured in the skeletal muscle and in isolated mitochondria of frogs at different stages during hibernation. We also measured the activity of lactate dehydrogenase (LDH) as an indicator of glycolytic capacity. The activities of cytochrome c oxidase, citrate synthase, and LDH were significantly lower in frog skeletal muscle after 4 mo of hibernation compared with control conditions. The reduction in skeletal muscle aerobic capacity is apparently due to changes in the intrinsic properties of the mitochondria. Overall, these results indicate an important reorganisation of ATP-producing pathways during long-term metabolic depression to match the lowered ATP demand.     

Exhaustive exercise training enhances aerobic capacity in American alligator (<Emphasis Type="Italic">Alligator mississippiensis</Emphasis>)

The oxygen transport system in mammals is extensively remodelled in response to repeated bouts of activity, but many reptiles appear to be ‘metabolically inflexible’ in response to exercise training. A recent report showed that estuarine crocodiles (Crocodylus porosus) increase their maximum metabolic rate in response to exhaustive treadmill training, and in the present study, we confirm this response in another crocodilian, American alligator (Alligator mississippiensis). We further specify the nature of the crocodilian training response by analysing effects of training on aerobic [citrate synthase (CS)] and anaerobic [lactate dehydrogenase (LDH)] enzyme activities in selected skeletal muscles, ventricular and skeletal muscle masses and haematocrit. Compared to sedentary control animals, alligators regularly trained for 15 months on a treadmill (run group) or in a flume (swim group) exhibited peak oxygen consumption rates higher by 27 and 16%, respectively. Run and swim exercise training significantly increased ventricular mass (~11%) and haematocrit (~11%), but not the mass of skeletal muscles. However, exercise training did not alter CS or LDH activities of skeletal muscles. Similar to mammals, alligators respond to exercise training by increasing convective oxygen transport mechanisms, specifically heart size (potentially greater stroke volume) and haematocrit (increased oxygen carrying-capacity of the blood). Unlike mammals, but similar to squamate reptiles, alligators do not also increase citrate synthase activity of the skeletal muscles in response to exercise.     

Growth, body composition and hepatic tyrosine aminotransferase activity in cortisol-fed channel catfish, Ictalurus punctatus Rafinesque

Metabolic consequences of chronic elevation of cortisol in the diet of yearling channel catfish, Ictalurus punctatus , were studied. Cortisol was incorporated into the diet in concentrations of 1, 10, 50 and 100 μg/g of food. This diet was offered at 3% of the body weight per day for 10 weeks. Fish were individually weighed and measured at 2-week intervals and feeding rates were adjusted. Body weight, liposomatic index and condition factor were significantly lower and food conversion was significantly higher in fish fed 50 or 100 μng cortisol/g of food when compared with controls. The hepatosomatic index of fish fed cortisol at the rate of 100 μg/g of food decreased significantly. Specific activity of hepatic tyrosine aminotransferase was significantly higher at the two highest cortisol doses. Long-term cortisol administration can reduce growth and condition factor by activating gluconeogenic mechanisms in which lipids and amino acids, rather than carbohydrates, are used for energy production. The metabolic effects of exogenous cortisol in this study offer an explanation for the decreased growth of fish under conditions that activate the secretion of endogenous cortisol.     

Critical thermal maxima of early life stages of three tropical fishes: Effects of rearing temperature and experimental heating rate

Marine ectotherms are often sensitive to thermal stress, and certain life stages can be particularly vulnerable (e.g., larvae or spawners). In this study, we investigated the critical thermal maxima (CT_max) of larval and early juvenile life stages of three tropical marine fishes (Acanthochromis polyacanthus, Amphiprion melanopus, and Lates calcarifer). We tested for potential effects of developmental acclimation, life stage, and experimental heating rates, and we measured metabolic enzyme activities from aerobic (citrate synthase, CS) and anaerobic pathways (lactate dehydrogenase, LDH). A slightly elevated rearing temperature neither influenced CT_max nor CS activity, which otherwise could have indicated thermal acclimation. However, we found CT_max to either remain stable (Acanthrochromis polyacanthus) or increase with body mass during early ontogeny (Amphiprion melanopus and Lates calcarifer). In all three species, faster heating rates lead to higher CT_max. Acute temperature stress did not change CS or LDH activities, suggesting that overall aerobic and anaerobic metabolism remained stable. Lates calcarifer, a catadromous species that migrates from oceanic to riverine habitats upon metamorphosis, had higher CT_max than the two coral reef fish species. We highlight that, for obtaining conservative estimates of a fish species’ upper thermal limits, several developmental stages and body mass ranges should be examined. Moreover, upper thermal limits should be assessed using standardized heating rates. This will not only benefit comparative approaches but also aid in assessing geographic (re-) distributions and climate change sensitivity of marine fishes.     

Activity of cytochrome c oxidase and lactate dehydrogenase in muscle tissue of slow growing (lower modal group) and fast growing (upper modal group) Atlantic salmon

Fast growing Atlantic salmon (upper modal group) exhibit a higher activity of muscle cytochrome c oxidase (CCO) and lactate dehydrogenase (LDH) than the slow growing salmon (lower modal group). The ratios of CCO/LDH activity, indicate a higher aerobic/anaerobic metabolic potential of the axial muscle in the upper modal group.