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.     

Pathway-specific response to cortisol in the metabolism of catfish

Cortisol produced biochemical pathway-specific effects on metabolic enzymes and other macromolecules in the freshwater catfish, Clarias batrachus. Injection of cortisol increased 1.6-fold activity of citrate synthase (CS) in brain, liver and skeletal muscle of the fish over vehicle-injected control, while administration of metyrapone (a cortisol synthesis inhibitor) reduced CS activity by 52%. Cortisol treatment of metyrapone-treated fish induced CS activity by approximately 2.5-fold, which was blocked after administration of actinomycin D or cycloheximide. This shows de novo synthesis of CS to enhance aerobic capacity of fish. In contrast the activities of glucose-6-phosphate dehydrogenase (G6-PDH) and lactate dehydrogenase (LDH) increased in response to metyrapone and decreased after administration of cortisol in all the three tissues. The cortisol-mediated decrease in G6-PDH and LDH activities reflects reduction in biosynthetic and anaerobic capacity of fish. Administration of metyrapone significantly increased RNA/DNA ratio and protein but cortisol decreased these macromolecular contents in brain, liver and skeletal muscle. It shows cortisol-induced decrease in protein synthesis capacity of fish. The present study suggests that cortisol-induces catabolic and aerobic but inhibits anabolic and anaerobic processes in freshwater catfish. The cortisol-dependent metabolic responses may also be associated with the permissive effect of cortisol on other hormone(s) in fish.     

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.     

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.     

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.     

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。       

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.     

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).     

Response of muscle-based biochemical condition indices to short-term variations in food availability in post-flexion reared sea bass Dicentrarchus labrax (L.) larvae

Six condition indices based on RNA, total soluble protein and two metabolic enzymes [lactate dehydrogenase (LDH) and citrate synthase (CS)] were analysed in muscle tissue of individual larvae, post-flexion reared sea bass Dicentrarchus labrax using DNA and total soluble protein as standards for size. In addition, the effect of 2 days of food deprivation on the cell proliferation rates was assessed. The RNA:DNA best reflected short-term changes in feeding conditions. If standardized by DNA content, LDH activity was a better indicator of condition than any other index but RNA:DNA. Further, the analysis of cell proliferation rates in muscle from 26 day-old larvae proved useful in distinguishing continuously fed larvae from individuals subjected to 2 days of fast.     

Histochemical observations on the occurrence of glycolytic and pentose phosphate cycle enzymes in the hepatopancreas and their possible relation to eyestalk factor(s) in the crab Scylla serrata (Forskal).

Histochemical studies were carried out on some of the glycolytic enzymes viz. phosphorylase, aldose, alpha-glycerophosphate dehydrogenase (alpha-GPDH) and lactic dehydrogenase (LDH) and a key enzyme of the pentose phosphatase cycle, glucose-6-phosphate dehydrogenase (G-6-PDH), in the hepatopancreas of Scylla serrata (Forskal). 1. Weak activities of phosphorylase and aldolase and strong-activities of alpha-GPDH and LDH were noticed mainly in the brush border of the tubules and R-cell cytoplasm. A trace activity of G-6-PDH was noticed in the brush border. 2. Bilateral eyestalk removal results in inhibition of both phosphorylase and aldolase. However, enhanced activities of alpha-GPDH and LDH were noticeable 4 h after the operation. The G-6-PDH activity remained unaltered till 24 h. 3. Injection of eyestalk extract into both intact and destalked crabs activated all the enzymes.     

Die Histotopik einiger Oxydoreduktasen im Hoden von Hund und Katze

Zusammenfassung Im Hoden von Hund und Katze werden folgende Enzyme histochemisch nachgewiesen: NADH-Tetrazoliumreduktase (NADH-T-Red), NADPH-Tetrazoliumreduktase (NADPH-T-Red), Cytochromoxydase (Cyt-Ox), Lactat-Dehydrogenase (LDH), Aldolase (ALD), Alkohol-Dehydrogenase (ADH), Glycerin-1-phosphat-Dehydrogenase (GDH), Glucose-6-phosphat-Dehydrogenase (G-6-PDH), Succinat-Dehydrogenase (SDH), NAD-spezifische Isocitrat-Dehydrogenase (NAD-ICDH). Die starke Fermentaktivität der G-6-PDH und der LDH in den Leydig-Zellen beider Spezies, der relativ hohe Gehalt an histochemisch nachweisbarer ADH in den Zwischenzellen der Katze sowie eine deutliche Reaktion auf GDH in den Sertoli-Zellen der Katze werden diskutiert.

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Summary In the testes of dog and cat the distribution pattern of NADH-tetrazolium reductase, NADPH-tetrazolium reductase, cytochrome oxydase, lactate dehydrogenase, aldolase, alcohol dehydrogenase, -glycerophosphate dehydrogenase, glucose-6-phosphate dehydrogenase, succinate dehydrogenase and NAD specific isocitrate dehydrogenase was studied by histochemical means. The strong reaction of G-6-PDH and LDH in the Leydig cells of both species, the relatively high amount of ADH in the interstitial cells of the cat testis and the principal site of -GPDH in the Sertoli cells of the cat are discussed.

     

Glycolytic enzymes in the premolt field crab Paratelphusa hydrodromus (Milne-Edwards) (Crustacea)

The quantitative assay of hexokinase (HK), phosphorylase, phosphofructokinase (PFK), glucose 6-phosphate dehydrogenase (G-6-PDH), glycerol 3-phosphate dehydrogenase (G-3 PDH) and lactate dehydrogenase (LDH) revealed that coxal muscles compared to hepatopancreas contained higher activities of all the enzymes investigated. It appears that the coxal muscles of the premolt field crab has carbohydrate-based fuel economy. The hepatopancreas is a rich source of lipid and very poor source of glycogen. The activity of G-6-PDH is moderately high in the hepatopancreas. It seems that in this lipogenic tissue conversion of G-6-P to triose phosphate occurs predominately via pentose-phosphate pathway thus generating NADPH for lipogenesis. The relative G-3PDH ad LDH activities in hepatopancreas and coxal muscles led us to believe that the reconversion of NAD from NADH in hepatopancreas nd muscle flexor is effected by glycerol 3-phosphate shuttle, whereas in muscle extensor it is achieved by both G-3PDH and LDH activities.     

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.     

Antitumor activities of iodoacetate and dimethylsulphoxide against solid Ehrlich carcinoma growth in mice

Treatment of tumor-bearing mice with LD12.5 values of iodoacetate; IAA (1.84 mg/100 g b.w.) and/or dimethylsulphoxide; DMSO (350 mg/100 g b.w.) significantly increased the cumulative mean survival time and percentage of survivors and reduced the mean tumor weight, compared to tumor-bearing controls, however, a more pronounced effect is recorded in the combined treatment. Also, an increase in the life span (ILS%) and tumor growth inhibition ratio (T/C%) are reported and amounted to 145.78 and 43.80%, 195.54 and 61.30% and 220.77 and 78.40% in IAA, DMSO and combined-treated groups, respectively. Results obtained from biochemical studies reveal that a single IAA treatment of tumor-bearing mice significantly increased the levels of plasma lactate dehydrogenase (LDH) activity, while it also significantly decreased the levels of plasma glucose and liver total protein, RNA and DNA, compared to normal controls. On the other hand, a single DMSO treatment significantly elevated the activities of blood antioxidant enzymes, i.e. glutathione peroxidase (GPx) and glucose-6-phosphate dehydrogenase (G6PDH) and decreased the liver RNA and DNA levels. Combined treatment increased significantly the levels of plasma LDH and erythrocytes G6PDH activities, as well as liver glycogen, and in contrast it decreased the levels of liver total protein, RNA and DNA, compared to normal controls.     

Na+--K+-adenosine triphosphatase and some oxidoreductases in the kidney of rats with spontaneous hypertension]

The activities of the Na+--K+-ATPase, succinic dehydrogenase (SDH/, lactic dehydrogenase (LDH/ and glucose-6-phosphat dehydrogenase (G-6-PDH/ were studied in the cortex outer and inner medulla of the kidneys of rats with spontaneous hypertension (SHR) and were compared with those of control normotensive Wistar rats. The SHR aged 6--8 weeks had durint the prehypertensive and the early hypertensive stage the same enzymatic activities as control rats. Rats with a steady SH aged 16-22 weeks had low specific activity of the, Na+--K+-ATPase, SDH and LDH in the outer medulla. The latter can be associated with decreased intensity of the energy metabolism and a reduction of the active sodium transport in the ascending limb of the loop of Henle in the SHR rats and cold cause the phenomenon of exaggerated natriuresis characteristic of hypertension.     

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.     

Metabolic differentiation of homologous leg muscles of two aquatic birds at the level of enzymatic organization

The quantitative determination of succinic dehydrogenase (SDH), hexokinase (HK), phosphorylase, phosphofructokinase (PFK), glycerol-3-phosphate dehydrogenase (G-3-PDH) and lactate dehydrogenase (LDH) was carried out in the homologous leg muscles of two aquatic Birds. It appears that the leg muscle fibres of the coot, a surface swimmer are more oxidative in nature and appear to utilize glucose as source of energy. The leg muscles of the dabchick, a diving Bird, on the other hand, seem to depend on glycogen as source of energy. The relative activity levels of HK, phosphorylase and PFK support the accepted r?le of glycogen as primary substrate of carbohydrate catabolism in the leg muscles. The ratio of G-3-PDH/LDH in the leg muscles revealed that glycerol 3-phosphate cycle appears to be insufficient to account for the major part of NADH oxidation. However, the LDH activity is quite high in all the muscles. These results led us to believe that glycerol 3-phosphate cycle may function during rest, when the rate of glycolysis will be low.     

Effect of tuftsin on the activity of energy metabolism enzymes in peritoneal macrophages

Cytochemistry was used to measure the activity of succinate dehydrogenase (SOD), lactate dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) in rat peritoneal macrophages under the action of the endogenous immunostimulant tuftcin (tre-lys-pro-arg) during phagocytosis of latex particles and at rest. Tuftcin did not affect the activity of the study enzymes in non-phagocytic cells. Elevation of the peptide concentration to 0.25 micrograms/ml and higher in phagocytic macrophages activated G-6-PDH and lowered the activity of LDH. Tuftcin did not alter the activity of SOD in phagocytic macrophages.     

The correlation between fish growth and several biochemical characteristics with reference to the steelhead <Emphasis Type="Italic">Parasalmo mykiss</Emphasis> Walb.

Correlation analysis demonstrated a statistically significant correlation of linear-weight characteristics of the steelhead rainbow trout (cultivated steelhead form) with an RNA/DNA ratio and the expression level of the gene encoding cytochrome c oxidase (CCO) in two-year-old individuals (1+) as well as the expression level of the gene encoding the myosin heavy chain (MyHC) and activities of the enzymes CCO and lactate dehydrogenase (LDH) in muscles and 1-glycerophosphate dehydrogenase (1-GPDH) and glucose-6-phosphate dehydrogenase (G6PDH) in the liver of two- and three-year-old individuals (1+ and 2+). With age, the correlation of 1-GPDH and G6PDH activities in the rainbow trout liver with the fish body weight increased, whereas their correlations with the body length reduced. The age- and sex-related distinctions in the MyHC gene expression and activities of the white muscle enzyme LDH and the liver enzymes 2-GPDH and G6PDH were detected in rainbow trout of both age cohorts.     

Glucose-6-phosphate dehydrogenase plays a pivotal role in nitric oxide-involved defense against oxidative stress under salt stress in red kidney bean roots

The pivotal role of glucose-6-phosphate dehydrogenase (G-6-PDH)-mediated nitric oxide (NO) production in the tolerance to oxidative stress induced by 100 mM NaCl in red kidney bean (Phaseolus vulgaris) roots was investigated. The results show that the G-6-PDH activity was enhanced rapidly in the presence of NaCl and reached a maximum at 100 mM. Western blot analysis indicated that the increase of G-6-PDH activity in the red kidney bean roots under 100 mM NaCl was mainly due to the increased content of the G-6-PDH protein. NO production and nitrate reductase (NR) activity were also induced by 100 mM NaCl. The NO production was reduced by NaN(3) (an NR inhibitor), but not affected by N(omega)-nitro-L-arginine (L-NNA) (an NOS inhibitor). Application of 2.5 mM Na(3)PO(4), an inhibitor of G-6-PDH, blocked the increase of G-6-PDH and NR activity, as well as NO production in red kidney bean roots under 100 mM NaCl. The activities of antioxidant enzymes in red kidney bean roots increased in the presence of 100 mM NaCl or sodium nitroprusside (SNP), an NO donor. The increased activities of all antioxidant enzymes tested at 100 mM NaCl were completely inhibited by 2.5 mM Na(3)PO(4). Based on these results, we conclude that G-6-PDH plays a pivotal role in NR-dependent NO production, and in establishing tolerance of red kidney bean roots to salt stress.