Hormonal control of glycerolphosphate dehydrogenase in the rat brain

—Following hypophysectomy or adrenalectomy, glycerolphosphate dehydrogenase (GPDH) (EC 1.1.1.8) activity decreased exponentially in the cerebral hemispheres and brain stem of adult male rats. The latter region was more affected than the former. Malate dehydrogenase (EC 1.1.1.40), isocitrate dehydrogenase (EC 1.1.1.42), lactate dehydrogenase (EC 1.1.1.27) and mitochondrial glycerolphosphate dehydrogenase (EC 1.1.95.5) activities remained unchanged. Injection of adrenocorticotrophic hormone or cortisol in hypophysectomized rats or cortisol in adrenalectomized rats restored GPDH activity. Thyroidectomy and gonadectomy had no effect on GPDH activity. Liver GPDH was not decreased by hypophysectomy or adrenalectomy. Muscle GPDH was diminished slightly by adrenalectomy and as much as brain GPDH by hypophysectomy. In young rats GPDH developmental increase in activity was inhibited by hypophysectomy. These results clearly show that brain GPDH activity is specifically regulated by cortisol (and probably closely related corticosteroids).     

Postnatal changes of some enzymatic activities of energy supplying metabolism in the cortex, inner and outer medulla of the rat kidney

1. In rat kidney cortex, outer and inner medulla the development of activities of seven enzymes was investigated during postnatal ontogeny (10, 20, 30, 60 and 90 days of age). The enzymes were selected in such a manner, as to characterize most of the main metabolic pathways of energy supplying metabolism: hexokinase (glucose phosphorylation, HK), glycerol-3-phosphate dehydrogenase (glycerolphosphate metabolism or shunt, GPDH), triose phosphate dehydrogenase (glycolytic carbohydrate breakdown, TPDH), lactate dehydrogenase (lactate metabolism, LDH), citrate synthase (tricarboxylic acid cycle, aerobic metabolism, CS), malate NAD dehydrogenase (tricarboxylic acid cycle, intra-extra mitochondrial hydrogen transport, MDH) and 3-hydroxyacyl-CoA-dehydrogenase (fatty acid catabolism, HOADH). 2. The renal cortex already differs metabolically from the medullar structures on the 10th day of life. It displays a high activity of aerobic breakdown of both fatty acids and carbohydrates. Its metabolic capacity further increases up to the 30th day of life. 3. The outer medullar structure is not grossly different from the inner medulla on the 10th day of life. Further it differentiates into a highly aerobic tissue mainly able to utilize carbohydrates. It can, however, to some extent, also utilize fatty acids aerobically and produce lactate from carbohydrates anaerobically. 4. The inner medullar structure is best equipped to utilize carbohydrates by anaerobic glycolysis, forming lactate. This feature is already pronounced on the 10th day of life, its capacity increases to some extent during postnatal development, being highest between the 10th and the 60th day of life.     

Regional enzyme development in rat brain. Enzymes of energy metabolism.

The development of several key enzymes of pyruvate and 3-hydroxybutyrate metabolism and of the tricarboxylic acid cycle was studied in six regions (cerebellum, medulla oblongata and pons, hypothalamus, striatum, mid-brain and cortex) of the neonatal, suckling and adult rat brain (2 days before birth to 60 days after birth). The enzymes whose developmental patterns were studied were: pyruvate dehydrogenase (EC 1.2.4.1), 3-hydroxybutyrate dehydrogenase (EC 1.1.1.30), citrate synthase (EC 4.1.3.7), NAD-linked isocitrate dehydrogenase (EC 1.1.1.41) and fumarase (EC 4.2.1.2). Citrate synthase, isocitrate dehydrogenase and pyruvate dehydrogenase develop as a cluster in each region, although the pyruvate dehydrogenase appears to lag slightly behind the others. As with the glycolytic-enzyme cluster [Leong & Clark (1984) Biochem. J. 218, 131-138] the timing of the development of the activity of this group of enzymes varies from region to region; 50% of the adult activity developed first in the medulla oblongata, followed by the hypothalamus, striatum and mid-brain, and then in the cortex and cerebellum respectively. The 3-hydroxybutyrate dehydrogenase activity also develops earlier in the medulla oblongata than in the other regions. The results are discussed with respect to the neurophylogenetic development of the brain regions studied and the importance of the development of the enzymes of aerobic glycolysis in relationship to the development of neurological maturation.     

ELECTROCONVULSIVE SEIZURE: AN INVESTIGATION INTO THE VALIDITY OF CALCULATING THE CYTOPLASMIC FREE [NAD+]/[NADH] [H+] RATIO FROM SUBSTRATE CONCENTRATIONS OF BRAIN

Abstract— This study is an investigation into the applicability of redox calculations to brain. At six intervals following electroconvulsive scizure, multiple metabolites were measured in freeze-blown brain from unanesthetized rats. From substrate ratios, the time course of the rapid changes in the cytoplasmic free [NAD⁺]/ [NADH] [H⁺] ratio was calculated from the reactions of lactate dehydrogenase [EC 1.1.1.27], malate dehydrogenase [EC 1.1.1.37] and glycerolphosphate dehydrogenase [EC 1.1.1.8], The pattern of the redox ratios in the control animals was also compared with the same ratios determined in freeze-clamped liver, a relatively homogeneous tissue. Though some evidence for effects of compartmentation are present in the results from brain, these effects are relatively minor. There was found to be very good agreement in the direction and magnitude of change of the redox ratios calculated from lactate dehydrogenase and malate dehydrogenase, and even from glycerolphosphate dehydrogenase at points of low flux. In spite of rapid changes of metabolites, the reaction catalyzed by glutamateoxaloacetate transaminase remained very near its equilibrium position at all time periods. From the results it has been concluded that in spite of the obvious structural heterogeneity of brain, meaningful calculations of the cytoplamic redox state in brain are possible.     

Development of glial cells in primary cultures: Energy metabolism and lactate dehydrogenase isoenzymes

Primary cultures of glial cells prepared from brains of newborn rats were grown for periods of 1–5 weeks. After a proliferative phase of between 2 and 3 weeks, the cultures were maintained in stationary phase, during which a significant increase of oxygen consumption and of the activities of lactate dehydrogenase, succinate dehydrogenase, and mitochondrial glycerolphosphate dehydrogenase could be observed. Furthermore, qualitative changes in the lactate dehydrogenase isoenzyme pattern were found with time, characterized by a shift toward an enhanced synthesis of H subunits. A similar development was found in comparing the LDH isoenzyme pattern in the brain of 15-day-old rat embryo with those of newborn and adult rat brains. It is suggested that some aspects of maturation of glial cells in culture are comparable to those occurring in whole brain in vivo, namely a shift towards an enhanced aerobic metabolism.     

Lactate dehydrogenase isoenzyme spectrum in various areas of the reticular formation of the brain in rats: the effect of emotional stress

The changes in lactate dehydrogenase (LDG) isoenzyme content in the various brain areas were studied in intact Wistar rats and upon immobilization stress. LDG fraction levels were compared to BP changes during immobilization. The proportion of "anaerobic" LDG fractions was higher and the proportion of "aerobic" fractions lower in the dorsal area of midbrain substantia reticularis than in medulla oblongata reticular formation. The changes in LDG fraction content related to BP alterations during immobilization were observed in dorsal and ventral areas of midbrain (but not medulla oblongata) substantia reticularis. The proportion of anaerobic LDG4 fraction in the dorsal area of midbrain substantia reticularis was higher in rats with hypertensive responses, than in hypotensive animals. The changes in LDG5 fraction content were opposite. In the ventral area of midbrain reticular formation BP reduction was accompanied by a significant rise in "anaerobic" and a decrease in "aerobic" LDG fraction levels. The data obtained indicate certain differences in the intensity of aerobic and anaerobic processes of carbohydrate degeneration in various areas of substantia reticularis in control rats, as well as the correlation of changes in energy metabolism in the brain with BP alterations during emotional stress.     

Activity of pyruvate- and ketoglutarate dehydrogenase complexes on various regions of the rat brain

The activity of pyruvate dehydrogenase and ketoglutarate dehydrogenase complexes (PDC; EC 1.2.4.1 and KDC; EC 1.2.4.2, respectively) was studied in extracts and lysates of mitochondria isolated from the cortex, cerebellum and stem of the rat brain. In all the mentioned cerebral areas the PDC activity calculating per 1 mg protein noticeably increased that for KDC. Under conditions of solubilization the activity of KDC lowered to a greater extent than that of PDC. The studied brain areas are arranged according to the activity of PDC and KDC in lysates in the following order: stem much much greater than cortex greater than cerebellum, however, the highest stock of the PDC activity manifesting with activation of endogenous phosphatase and dephosphorylation of the complex is observed in cerebellum. When calculating per mitochondria isolated from 1 g of tissue, the value of the PDC activity/KDC activity ratio in all areas of the brain is more than 1, and in cerebellum allowing for the complete PDC activity it exceeds 3. The data obtained emphasize the biochemical originality of the cerebellum and the presence of specific peculiarities in regulation of the pyruvate dehydrogenase complex activity in this cerebral area.     

Time-dependence of the effect of X-irradiation on the formation of glycerol phosphate dehydrogenase and other dehydrogenases in the developing rat brain

X-irradiation (100-1500 r) administered to the heads of rats 8-30 days of age inhibited the development of glycerol phosphate dehydrogenase (l-glycerol 3-phosphate-NAD oxidoreductase, EC 1.1.1.8) in the brain stem and cerebral hemispheres. At 40 days of age and older no effect was observed. This inhibition was a delayed phenomenon, dose-dependent and with no recovery. It is proposed that the inhibition of enzyme formation is related to radiation damage caused to DNA. Actinomycin D inhibited the development of glycerol phosphate dehydrogenase in a manner similar to ionizing radiation. Four other dehydrogenases also showed age-dependent radiosensitivities. ;Malic enzyme' (EC 1.1.1.40), lactate dehydrogenase (EC 1.1.1.27) and malate dehydrogenase (EC 1.1.1.37) ceased to be radiosensitive at about 8 days of age and isocitrate dehydrogenase (NADP) (EC 1.1.1.42) at 16 days. The correlation between developmental increase in enzyme activity and radiosensitivity held closely for glycerol phosphate dehydrogenase and isocitrate dehydrogenase and to a smaller extent for the others.     

Some properties of lactate dehydrogenase found in human urine

1. Urinary lactate dehydrogenase was concentrated and subjected to starchblock electrophoresis. The isoenzyme pattern obtained was shown to be similar to that of the kidney cortex and medulla but different from that of the bladder and kidney pelvis. 2. The values for the relative activity and Michaelis constant of urinary lactate dehydrogenase were similar to those for kidney cortex and medulla but significantly different from those obtained for bladder and kidney pelvis. 3. The molecular weight of urinary lactate dehydrogenase was estimated by thin-layer chromatogtaphy on Sephadex G-200. The values obtained for several samples of urine ranged from 129 000 to 155 000 and were very close to that of the crystalline rabbit-muscle enzyme (140 000). 4. The question of the possible origin of urinary lactate dehydrogenase is discussed and the conclusion drawn that the kidney and not the plasma is the most likely source.     

Control of aerobic glycolysis in the brain in vitro

Protoveratrine-(5 M) stimulated aerobic glycolysis of incubated rat brain cortex slices that accompanies the enhanced neuronal influx of Na⁺ is blocked by tetrodotoxin (3 M) and the local anesthetics, cocaine (0.1 mM) and lidocaine (0.5 mM). On the other hand, high [K⁺]-stimulated aerobic glycolysis that accompanies the acetylcholine-sensitive enhanced glial uptakes of Na⁺ and water is unaffected by acetylcholine (2 mM). Experiments done under a variety of metabolic conditions show that there exists a better correlation between diminished ATP content of the tissue and enhanced aerobic glycolysis than between tissue ATP and the ATP-dependent synthesis of glutamine. Whereas malonate (2 mM) and amino oxyacetate (5 mM) suppress ATP content and O₂ uptake, stimulate lactate formation, but have little effect on glutamine levels, fluoroacetate (3 mM) suppresses glutamine synthesis in glia, presumably by suppressing the operation of the citric acid cycle, with little effect on ATP content, O₂ uptake, and lactate formation. Exogenous citrate (5 mM), which may be transported and metabolized in glia but not in neurons, inhibits lactate formation by cell free acetone-dried powder extracts of brain cortex but not by brain cortex slices. These results suggest that the neuron is the major site of stimulated aerobic glycolysis in the brain, and that under our experimental conditions glycolysis in glia is under lesser stringent metabolic control than that in the neuron. Stimulation of aerobic glycolysis by protoveratrine occurs due to diminution of the energy charge of the neuron as a result of stimulation of the sodium pump following tetrodotoxin-sensitive influx of Na⁺; stimulation by high [K⁺, NH₄ ⁺, or Ca²⁺ deprivation occurs partly by direct stimulation of key enzymes of glycolysis and partly by a fall in the tissue ATP concentration.     

Schistosomatium douthitti: carbohydrate metabolism in the adults.

Pathways of carbohydrate metabolism in the adults of Schistosomatium douthitti: were investigated. Histochemical reactions for adenosinetriphosphatase (EC 3.6.1.3) glucose 6-phosphate dehydrogenase (EC 1.1.1.49), phosphogluconate dehydrogenase (EC 1.1.1.43), glycerol-3-phosphate dehydrogenase (EC 1.1.1.8), lactate dehydrogenase (EC 1.1.1.27, 1.1.2.3) isocitrate dehydrogenase (EC 1.1.1.41), succinate dehydrogenase (EC 1.3.99.1), malate dehydrogenase (EC 1.1.1.37), cytochrome oxidase (EC 1.9.3.1), and adenosine triphosphatase (EC 3.6.1.3) were found in the adult worms. Glycogen deposits occurred in the parenchyma.Low oxygen tension immobilized the worms. Tartar emetic, sodium cyanide reduced adult motility in vitro. Manometric experiments demonstrated a respiratory quotient of approximately one. Oxygen uptake was completely inhibited by tartar emetic and partially inhibited by sodium fluoracetate and sodium cyanide. Inhibition by sodium fluoroacetate was partially counteracted by citric acid in the medium.Adults demonstrated an oxygen debt following anaerobic incubation. A maximum of 52% of the glucose consumed under aerobic conditions was excreted as lactic acid. Under anaerobic conditions the amount of lactic acid excreted increased. Acids other than lactic acid were also released. Results indicate that although glycolysis is the major pathway, two additional aerobic pathways also exist, one which is cyanide sensitive and the other cyanide insensitive.     

The Activities of Some Energy-Metabolising Enzymes in Nonsynaptic (Free) and Synaptic Mitochondria Derived from Selected Brain Regions

Abstract: The enzyme complement of two different mitochondrial preparations from adult rat brain has been studied. One population of mitochondria (synaptic) is prepared by the lysis of synaptosomes, the other (nonsynaptic or free) by separation from homogenates. These populations have been prepared from distinct regions of the brain: cortex, striatum, and pons and medulla oblongata. The following enzymes have been measured: pyruvate dehydrogenase (EC 1.2.4.1), citrate synthase (EC 4.1.3.7), NAD-linked isocitrate dehydrogenase (EC 1.1.1.41), NADP-linked isocitrate dehydrogenase (EC 1.1.1.42), fumarase (EC 4.2.1.2), NAD-linked malate dehydrogenase (EC 1.1.1.37), D-3-hydroxybutyrate dehydrogenase (EC 1.1.1.30), and mitochondrially bound hexokinase (EC 2.7.1.1) and creatine kinase (EC 2.7.3.2). The nonsynaptic (free) mitochondria show higher enzyme specific activities in the regions studied than the corresponding values recorded for the synaptic mitochondria. The significance of these observations is discussed in the light of the different metabolic activities of the two populations of mitochondria and the compartmentation of the metabolic activities of the brain.     

Protein S100B release from rat brain slices during and after ischemia: comparison with lactate dehydrogenase leakage

One hour of ischemia significantly increased protein S100B release from rat brain slices without altering lactate dehydrogenase leakage. Reoxygenation of the ischemic slices, however, increased the levels of these biochemical markers in the medium. Although removal of extracellular Ca⁺² ions from the medium did not alter the basal lactate dehydrogenase leakage from cortical slices, an excessive increase in basal protein S100B release was seen under this condition. Ischemia and/or reoxygenation induced enhancements in these markers were attenuated by removal of Ca⁺² ions from the medium. Ischemia significantly increased glutamate release, but neither ischemia nor reoxygenation induced rises in protein S100B and lactate dehydrogenase levels were altered by glutamate receptor antagonists. Rising the glutamate levels in the medium by each ouabain or exogenous glutamate, moreover, failed in exerting an ischemia like effect on protein S100B and LDH outputs. In contrast, exogenous glutamate added into the medium protected the slices against reoxygenation induced increments in protein S100B and lactate dehydrogenase levels.

These results indicate that protein S100B has a greater sensitivity against ischemia than lactate dehydrogenase in in vitro brain slice preparations. Since neither exogenous glutamate nor enhancements of the extracellular glutamate levels by ouabain had an ischemia like effect, and since glutamate receptor antagonists were also unsuccessful, it seems unlikely that ischemia-induced increase in glutamate release is directly involved in protein S100B release or lactate dehydrogenase leakage determined in the present study.     

AMINO ACID METABOLISM AND AMMONIA FORMATION IN BRAIN SLICES

The formation of ammonia and changes in the contents of free amino acids have been investigated in slices of guinea pig cerebral cortex incubated under the following conditions: (1) aerobically in glucose-free saline; (2) aerobically in glucose-free saline containing 10 mM-bromofuroic acid, an inhibitor of glutamate dehydrogenase (EC 1.4.1.2); (3) aerobically in saline containing 11-1 mM-glucose and (4) anaerobically in glucose-free saline. Ammonia was formed at a steady rate aerobically in glucose-free medium. The formation of ammonia was largely suppressed in the absence of oxygen or in the presence of glucose whereas the inhibitor of glutamate dehydrogenase produced about 50 per cent inhibition. Other inhibitors of glutamate dehydrogenase exerted a similar effect. Ammonia formation was also inhibited by some inhibitors of aminotransferases but not by others. Inhibition was generally more pronounced during the second and third hour of incubation. With the exception of glutamine which decreased slightly, the contents of all amino acids increased markedly during the anaerobic incubation. During aerobic incubation in a glucose-free medium, there was an almost complete disappearance of glutamic acid and GABA. Glutamine also decreased, but to a relatively smaller extent. The content of all other amino acids increased during aerobic incubation in glucose-free medium, although to a lesser extent than under anaerobic conditions. The greater increase of amino acids appearing anaerobically in comparison to the increase or decrease occurring under aerobic conditions corresponded closely to the greater amount of ammonia formed aerobically over that formed anaerobically. This finding is interpreted as indicating a similar degree of proteolysis under anaerobic and aerobic conditions; aerobically, the amino acids are partly metabolized with the concomitant liberation of ammonia. In glucose-supplemented medium, the content of glutamine was markedly increased. The content of glutamate and aspartate remained unchanged, whereas that of some other amino acids increased but to a lesser extent than in the absence of glucose. Proteolysis in the presence of glucose was estimated at about 65 per cent of that in its absence. In the presence of bromofuroate the rate of disappearance of glutamate was unchanged, but there was a larger increase in the content of aspartate and a smaller decrease of GABA and glutamine. Other changes did not differ significantly from those observed in the absence of bromofuroate. We conclude that the metabolism of amino acids in general and of glutamic acid in particular differs according to whether they are already present within the brain slice or are added to the incubation medium. Only the endogenous amino acids appear to be able to serve as precursors of ammonia and as substrates for energy production.     

Evaluation of the rate-limiting steps in the pathway of glucose metabolism in kidney cortex of normal, diabetic, cortisone-treated and growth hormone-treated rats

1. The activities of gluconeogenic and glycolytic enzymes and the concentrations of citrate, ammonia, amino acids, glycogen, glucose 6-phosphate, acetyl-CoA, lactate and pyruvate were measured in kidney cortex of normal, diabetic, cortisone-treated and growth hormone-treated rats. 2. In kidney cortex of diabetic, cortisone-treated and growth hormone-treated rats the activities of glucose 6-phosphatase (EC 3.1.3.9), fructose 1,6-diphosphatase (EC 3.1.3.11) and phosphopyruvate carboxylase (EC 4.1.1.32) were increased. 3. The activities of glutamate dehydrogenase (EC 1.4.1.3), alanine aminotransferase (EC 2.6.1.2), aspartate aminotransferase (EC 2.6.1.10) and pyruvate carboxylase (EC 6.4.1.1) were increased in diabetic and cortisone-treated rats. In growth hormone-treated rats the activity of aspartate aminotransferase was depressed but those of the other three enzymes were unchanged. 4. The activity of hexokinase (EC 2.7.1.1) was not altered in any of these conditions. Phosphofructokinase (EC 2.7.1.11) activity was depressed only in growth hormone-treated rats. Pyruvate kinase (EC 2.7.1.40) activity was depressed in cortisone-treated and growth hormone-treated rats but unchanged in diabetic rats. 5. Amino acids, acetyl-CoA and glucose 6-phosphate contents were increased in rat kidneys in all these three conditions. Ammonia content was increased in diabetic and cortisone-treated rats but was markedly diminished in growth hormone-treated rats. 6. The [lactate]/[pyruvate] ratio was elevated in diabetic and cortisone-treated rats but unchanged in growth hormone-treated rats. Citrate content was increased in the kidney cortex of diabetic and growth hormone-treated rats but was unchanged in cortisone-treated rats. The activity of ATP citrate lyase (EC 4.1.3.8) was depressed in diabetic and growth hormone-treated rats but was increased in cortisone-treated rats. 7. Glycogen content was moderately elevated in growth hormone-treated rats and markedly elevated in diabetic rats, whereas no change in glycogen content was observed in cortisone-treated rats. Glycogen synthetase (EC 2.4.1.11) activity was unchanged in all these three conditions. Phosphorylase (EC 2.4.1.1) activity was not affected in cortisone-treated rats but was depressed in diabetic and growth hormone-treated rats.     

Cerebral carbohydrate metabolism during acute hypoxia and recovery

Abstract— The levels of ATP, ADP, AMP and phosphocreatine, of four amino acids, and of 11 intermediates of carbohydrate metabolism in mouse brain were determined after: (1) various degrees of hypoxia; (2) hypoxia combined with anaesthesia; and (3) recovery from severe hypoxia. Glycogen decreased and lactate rose markedly in hypoxia, but levels of ATP and phosphocreatine were normal or near normal even when convulsions and respiratory collapse appeared imminent. During 30 s of complete ischaemia (decapitation) the decline in cerebral ATP and phosphocreatine and the increase in AMP was less in mice previously rendered hypoxic than in control mice. From the changes we calculated that the metabolic rate had decreased by 15 per cent or more during 30 min of hypoxia. Hypoxia was also associated with decreases of cerebral 6-phosphogluconate and aspartate, and increases in alanine, γ-aminobutyrate, α-ketoglutarate, malate, pyruvate, and the lactate :pyruvate ratio. Following recovery in air (10 min), increases were observed in glucose (200 per cent), glucose-6-phosphate, phosphocreatine and citrate, and there was a fall in fructose-1, 6-diphosphale. Similar measurements were made in samples from cerebral cortex, cerebellum, midbrain and medulla. Severe hypoxia produced significant increases in lactate and decreases in glycogen in all areas; γ-aminobutyrate levels increased in cerebral cortex and brain stem, but not in cerebellum. No significant changes occurred in ATP and only in cerebral cortex was there a significant fall in phosphocreatine. Phosphocreatine, ATP and glycogen were determined by quantitative histochemical methods in four areas of medulla oblongata, including the physiological respiratory centre of the ventromedial portion. After hypoxia, ATP was unchanged throughout and the changes (decreases) in phosphocreatine and glycogen were principally confined to dorsal medulla, notably the lateral zone. Thus there is no evidence that respiratory failure is caused by a ‘power’ failure in the respiratory centre. It is suggested that in extremis a protective mechanism may cause neurons to cease firing before high-energy phosphate stores have been exhausted.     

Properties and Regional Distribution of Pyruvate Dehydrogenase Kinase in Rat Brain

A method is described to measure directly in rat brain the activity of pyruvate dehydrogenase kinase (PDHa kinase; EC 2.7.1.99), which catalyzes the inactivation of pyruvate dehydrogenase complex (PDHC, EC 1.2.4.1, EC 2.3.1.12, and EC 1.6.4.3). The activity showed the expected dependence on added ATP and divalent cation, and the expected inhibition by dichloroacetate, pyruvate, and thiamin pyrophosphate. These results, and the properties of pyruvate dehydrogenase phosphate phosphatase (EC 3.1.3.43), indicate that the mechanisms of control of phosphorylation of PDHC seem qualitatively similar in brain to those in other tissues. Regionally, PDHa kinase is more active in cerebral cortex and hippocampus, and less active in hypothalamus, pons and medulla, and olfactory bulbs. Indeed, the PDHa kinase activity in olfactory bulbs is uniquely low, and is more sensitive to inhibition by pyruvate and dichloroacetate than that in the cerebral cortex. Thus, there are significant quantitative differences in the enzymatic apparatus for controlling PDHC activity in different parts of the brain.     

BIOCHEMICAL CHANGES IN THE BRAIN IN RATS POISONED WITH AN ALKYLMERCURY COMPOUND, WITH SPECIAL REFERENCE TO THE INHIBITION OF PROTEIN SYNTHESIS IN BRAIN CORTEX SLICES

The incorporation of [U-¹⁴C]leucine into the protein of brain cortex slices from rats poisoned with methylmercury thioacetamide was markedly inhibited before the development of neurological symptoms and when the oxygen consumption, aerobic and anaerobic glycolysis and sulphydryl enzyme activities were unchanged. After the appearance of neurological symptoms, the oxygen consumption decreased significantly, while lactic acid formation did not change under anaerobic conditions, but slightly decreased under aerobic conditions. The activities of the three sulphydryl enzymes (Mg-activated ATPase, fructose- diphosphate aldolase and succinate dehydrogenase) were almost the same in visual cortex, motor cortex, cerebellum and caudate nucleus, while the activities of Mg-activated ATPase and succinate dehydrogenase in the white matter were lower than that in the grey matter. There was no difference in the activity of fructosediphosphate aldolase in grey and white matter. The activities of all three enzymes did not show any change in the earlier stage of poisoning when the animal remained free from neurological symptoms. At the more advanced stage, when neurological symptoms were present, only the activity of the succinate dehydrogenase decreased significantly, while the activities of the other two enzymes remained unchanged. The selective inhibition of protein synthesis may have a direct bearing on the poisoning by the alkylmercury compound.     

The effect of food deprivation on enzyme activity in developing brain

Brain and body weights, contents of DNA and protein and activities of 1,6-diphosphofructoaldolase (aldolase, EC 4.1.2.13), creatine phosphokinase (CPK, EC 2.7.3.2), and isocitric dehydrogenase (ICD, EC 1.1.1.42) in brain (minus cerebellum and brain stem) were studied in control and food-deprived rats at 7, 14 and 21 days of postnatal age. Activities of all three enzymes per brain were less in the food-deprived animals. In both groups of rats the ratios of aldolase/DNA and CPK/DNA increased with maturation, indicating that increasing activity per brain during maturation was the result of both increased activity per cell and increased numbers of cells. The ratio of ICD/DNA decreased with maturation but was essentially the same in both the food-deprived and control groups. Increase of ICD activity per brain with maturation was attributable to increased numbers of cells. Food deprivation in immature animals resulted in lowered activities per brain for aldolase, CPK and ICD because of diminished cell multiplication.     

Some neurochemical consequences of repeated ethanol loading in rat brain

Neurochemical consequences of repeated ethanol treatment on energy and ammonia metabolism were studied in different regions of rat brain. Energy production was decreased as indicated by lowered lactate dehydrogenase and succinate dehydrogenase activities with possible lacticacidimia. Transamination of alanine and aspartate increased while the deamination of glutamate decreased in all the regions of brain. The deamination of AMP was slightly elevated in cerebral cortex and brain stem while it was inhibited in cerebellum. Ammonia levels were persistently high, despite stepped up glutamine synthesis and ureogenesis. The synergistic action of ammonia during ethanol intoxication is envisaged.