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1.
Abstract— [2-14C]Propionate injected into rats was metabolized into [14C]glucose and 14C-labelled aspartate, glutamate, glutamine and alanine. The results are consistent with the conversion of propionate into succinate and the oxidation of succinate into oxaloacetate, the precursor of labelled amino acids and the substrate for gluconeogenesis.
The ratio of the specific radioactivity of glutamine to glutamate was greater than 1 during the 30 min period in the brain, indicating that propionate taken up by the brain was metabolized mainly in the 'small glutamate compartment' in the brain. The results, therefore, support the previous conclusion (G aitonde , 1975) that the labelling of amino acids by [14C]propionate formed from [U-14C>]-threonine in thiamin-deficient rats was metabolized in the 'large glutamate compartment' of the brain.
The specific radioactivity ratio of glutamine to glutamate in the liver was less than 1 during the 10 min period but greater than 1 at 30min. These findings which gave evidence against metabolic compartments of glutamate in the liver, were interpreted as indicative of the entry of blood-borne [14C]glutamine synthesized in other tissues, e.g. brain. The labelling of amino acids when compared to that after injection of [U-14C]glucose showed that [2-14C]propionate was quantitatively a better source of amino acids in the liver. The concentration of some amino acids in the brain and liver was less in the adult than in the young rats, except for alanine and glutathione, where the liver content was more than double that in the adult.  相似文献   

2.
Abstract— [U-14C]Ribose was given by subcutaneous injection to young rats aged 2–56 days. During the first week after birth 14C in the brain was found mainly combined in glucose, fructose and sedoheptulose which contained 46–57 per cent of the 14C in the acid soluble metabolites in the rat brain. In contrast, during the critical period (10–15 days after birth) the 14C in the free sugars decreased from 24 to 3 per cent, while the 14C content of amino acids in the brain increased from 11 to 44 per cent of the total perchloric acid-soluble 14C. The increase in labelling of amino acids during the critical period was attributed to increased glycolysis and increased oxidation of pyruvate. The relative specific radioactivity of y -aminobutyrate and aspartate in the rat brain at 28 days after birth was equal to or greater than the relative specific radioactivity of glutamate. Assuming that the increase in amino acid content following the cessation of cell proliferation in the brain is located mainly in cell processes (cytoplasm of axons, dendrites, glial processes and nerve terminals), tentative values were estimated for the pool sizes of glutamate, glutamine, aspartate and y -amino butyrate.  相似文献   

3.
Abstract— The incorporation of 14C into amino acids of the brain was determined at different times after injection of [U-14C]glucose and [U-14C]ribose to rats maintained on thiamine-supplemented and thiamine-deficient diets for 22 days.
The 14C-content of amino acids in the brain of thiamine-deficient rats decreased at times 2–10 min after injection of [U-14C]glucose. but it increased at 2 min and decreased at times 5–10 min after injection of [U-14C]ribose.
The results of labelling of amino acids indicated that the activities in vivo of the thiamine pyrophosphate requiring enzymes, pyruvate oxidase, a-oxoglutarate dehydrogenase and transketolase were similar in the two groups. It was suggested that the observed decrease in the labelling of amino acids was due to one or more of the following factors: (i) a decrease in the activities of glycolytic enzymes catalysing the conversion of glucose into triose phosphate; (ii) a decrease in the transport of substrate to the active site of the enzymes; or (iii) altered neurohistopathology of the brain.
Thiamine deficiency in rats showed a 5% decrease in glutamate ( P < 0–05), 46% decrease in threonine (P < 0001) and 16% increase in glycine ( P < 0–01) content of the brain.  相似文献   

4.
Previous studies have shown that rainbow trout fed on diets containing whole protein have superior growth rates compared to fish fed on diets of similar amino acid composition but containing a high proportion of free amino acids. The influence of several nutritional factors on the uptake of radioactivity from food pellets containing either [U-I4C] protein or [U-14C] amino acids into the systemic blood of trout has been investigated. The time taken for radioactivity in the free amino acid fraction of blood to reach a peak after a meal containing [U-14C] protein had been given was much shorter, and the level of radioactivity in the blood higher, in trout with almost empty stomachs than in fish with almost full stomachs; uptake of radioactivity into blood amino acids was also more rapid and reached much higher concentrations when pellets containing [U-14C] amino acids were fed than when [U-14C] protein was fed. Incorporation of radioactivity into blood protein continued for a much longer period and reached higher levels when a pellet containing [U-14C] protein was fed than when a pellet containing [U-14C] amino acids was fed. Previous dietary history (low or high protein intake) did not appear to affect the rate of absorption of amino acids from either protein or free amino acid pellets. The uptake rates from pellets containing free amino acids could be slowed by mixing the dietary amino acids with albumin. The distribution, postabsorption, of radioactivity in the different fractions of blood and liver suggested that incorporation of carbon residues into glycogen and lipid from an amino acid diet was greater than from a protein diet. The converse was true of incorporation of radioactivity into tissue protein.  相似文献   

5.
IN VIVO INHIBITION OF RAT BRAIN PROTEIN SYNTHESIS BY l-DOPA   总被引:3,自引:2,他引:1  
Abstract— A study has been made of the effect of a single intraperitoneal dose of l -DOPA on the in vivo metabolism of [14C]leucine and [14C]lysine by the brain, and on their uptake into brain protein. Administration of 500 mg DOPA/kg to 40-g rats raised the concentrations of several free amino acids; the only amino acid which underwent a statistically significant increment was alanine. Intracisternally-injected [U-14C]leucine was rapidly metabolized to other labelled compounds; DOPA administration did not influence significantly the rate of its metabolism. No similar metabolic change was observed after administering [U-14C]lysine intracisternally.
Incorporation of [14C]leucine and [14C]lysine into total brain protein was significantly reduced 45 min after DOPA administration. There was also depression of the uptake of labelled amino acid into a supernatant fraction, obtained by high speed centrifugation of the brain homogenate, and into brain microtubular protein (tubulin). Reduced amino-acid incorporation into brain proteins observed 45 min after l -DOPA injection coincided with extensive disaggregation of brain polyribosomes. At 120 min after DOPA treatment, disaggregation was no longer significant and there was a smaller depression in labelled amino aicd incorporation, which disappeared completely 240 min after l -DOPA injection. It is concluded that disaggregation of brain polysomes following DOPA treatment is an accurate reflection of a change in the intensity of brain protein synthesis in vivo.  相似文献   

6.
Abstract— The characteristics of the uptake of l -[U-14C] glutamate into rat dorsal sensory ganglia were investigated. The uptake was mediated by two distinct kinetic systems, with apparent Km values of the order of 10−3 M (low affinity) and 10−5 m (high affinity). The high affinity uptake system was strongly dependent upon temperature and sodium ion concn, and was depressed by a number of metabolic inhibitors. Following uptake, [14C] glutamate was extensively metabolized, primarily to glutamine, although this was not so with cultured ganglia, where in addition to an increased uptake of [14C] glutamate, the specific radioactivity of glutamate was increased and that of glutamine decreased. The labelled substrates [U-14C]pyruvate and [U-14C] acetate were used to investigate this phenomenon and the results are discussed in relation to current knowledge of metabolic compartmentation in nervous tissue.  相似文献   

7.
Abstract. The fate of radioactively labelled amino acids injected into the haemolymph of the aphid Aphis fabae was investigated. Radioactivity from each of L-[U-14C]-glutamic acid, L-[U-14C]-serine and L-[U-14C]-threonine in the aphid tissues declined exponentially, at rates of 32, 9.3 and 1.0 pmol/aphid/min, respectively. For 14C-glutamic acid, radioactivity lost from the aphids was recovered quantitatively as carbon dioxide, and radioactivity in aphid saliva and honeydew was undetectable. When expressed on a per unit aphid biomass basis, the rate of respiratory loss of glutamic acid from aphids reared on chemically-defined diets was more than double that of aphids reared on the host plant, Vicia faba . It is concluded that respiration is a quantitatively important component to the aphid metabolism of glutamic acid and other amino acids.  相似文献   

8.
Abstract— Changes in morphology and in transformations of [U-14C]glucose and [1-14C]acetate into amino acids of the brain cortex were followed on the Sth, 10th and 21st days after production of mechanical lesions and compared with control tissue. In the experimental tissue, proliferation of astroglia and reduction of the number of neurons had taken place. On the 10th day, accumulation of mitochondria and of some gliofilaments in the cytoplasm of astroglia was observed. On the 21st day, the gliofilaments occupied a substantial portion of the astroglial cytoplasm and the mitochondria were reduced in number and compressed to the cell membrane. Incorporation of 14C from acetate into amino acids was substantially increased on the 10th day (up to 240% with respect to controls) and normalized again on the 21st day. Incorporation of [14C]glucose into amino acids decreased somewhat during the experimental period. It has been proposed that the proliferation of astrocytes and their ultrastructural changes may account for the increased transformation of [14C]acetate into amino acids, in particular into glutamine which is formed from the small glutamate pool.  相似文献   

9.
Abstract: Chains of lumbar sympathetic ganglia from 15-day-old chicken embryos were incubated for 4 h at 36°C in a bicarbonate-buffered salt solution equilibrated with 5% CO2-95% O2. Glucose (1–10 m M ), lactate (1–10 m M ), [U-14C]glucose, [1-14C]glucose, [6-14C]glucose, and [U-14C]lactate were added as needed. 14CO2 output was measured continuously by counting the radioactivity in gas that had passed through the incubation chamber. Lactate reduced the output of CO2 from [U-14C]glucose, and glucose reduced that from [U-14C]lactate. When using uniformly labeled substrates in the presence of 5.5 m M glucose, the output of CO2 from lactate exceeded that from glucose when the lactate concentration was >2 m M . The combined outputs at each concentration tested were greater than those from either substrate alone. The 14CO2 output from [1-14C]glucose always exceeded that from [6-14C]glucose, indicating activity of the hexose monophosphate shunt. Lactate reduced both of these outputs, with the maximum difference between them during incubation remaining constant as the lactate concentration was increased, suggesting that lactate may not affect the shunt. Modeling revealed many details of lactate metabolism as a function of its concentration. Addition of a blood-brain barrier to the model suggested that lactate can be a significant metabolite for brain during hyperlactemia, especially at the high levels reached physiologically during exercise.  相似文献   

10.
Abstract: The incorporation of amino acids into brain proteins following brachial plexus stimulation (BPS) was studied in anaesthetised Sprague-Dawley rats following injection of radioactive precursors of both neuronal and glial compartments. Following intraperitoneal injection of [14C]glucose, which is the major neuronal pool precursor, BPS resulted in a significant increase of 379% ( P ± 0.001) in the incorporation of carbon from [14C]glucose into TCA-insoluble proteins in the contralateral sensorimotor cortex as compared with the ipsilateral area of the same animal. This increase was abolished totally when tetrodotoxin (10 μg ml-1) was applied topically to the surface of the stimulated area. Following intraperitoneal injection of [14C]acetate, which is considered to be mainly a glial cell precursor, the same afferent electrical stimuli caused a significant decrease of 21% in the incorporation of amino acids into proteins in the stimulated versus unstimulated sensorimotor cortex. With [4-3H]phenyl-alanine or [l-14C]leucine as precursors a significant decrease (12%) or no change was recorded, respectively. A similar decrease in protein synthesis in the stimulated sensorimotor cortex was achieved using different routes of injection. No significant changes were observed in the ratio of the specific radioactivities of the total amino acids of the two hemispheres using either precursor. In vitro , synaptosomes showed a large increase in incorporation into proteins after treatment with electrical pulses, both with [14C]glucose and with [U-14C]acetate as precursors.  相似文献   

11.
Abstract— Mammalian cortical synaptosomes incubated in the presence of glucose (2.5 MM) plus glutamine (0.5 mM) showed a 30% increase in transmitter amino acid content over controls with glucose alone and a doubling of glutamate release induced by Veratrine or high K+. Double-label experiments, i.e. [U-14C]glucose with [3H]glutamine, and single-label experiments, i.e. [U-14C]glucose or [U-14C]-glutamine showed that stimulus-released glutamate was derived principally (80%) from glutamine. Released glutamine-derived glutamate was of higher (x 2) specific radioactivity than its tissue equivalent. Glutamine alone (0.5–0.75 mM) was much less effective than equivalent amounts of glucose alone, in stimulating respiration and maintaining tissue K+ levels.  相似文献   

12.
Abstract— The effect of 15 h continuous exposure to CS2 on the metaboliam of glucose and free amino acids in the brain of rats was studied. CS2 caused a moderate hypoglycaemia. There were also changes in the amounts of some amino acids in the brain. Glutamate and γ-aminobutyrate were lower whereas glutamine was markedly increased. Comparative studies in vivo of the metabolism of [2-14C]glucose and [1-14C]butyrate indicated that CS2 did not affect glycolysis or the incorporation of 14C from glucose into amino acids except into γ-aminobutyrate which was reduced. Contrary to the findings with [14C]glucose, CS2 provoked distinct changes in the labelling of amino acids when [14C]butyrate was the precursor. The most notable change was a markedly increased incorporation of 14C into glutamine. Based on the two-compartment model of brain glutamate the experimental findings indicated that CS2 affected metabolism associated with the 'small' pool of glutamate but had a minimal effect on metabolism associated with the 'large' glutamate pool. The possibility is suggested that the changes observed involved an increased rate of ammonia removal. The low incorporation of 14C into γ-aminobutyrate from either precursor is consistent with other evidence showing that CS2 interferes with pyridoxal phosphate-dependent enzymes.  相似文献   

13.
Abstract— The metabolism of γ-hydroxybutyrate (GHB) was studied by following the fate of [1-14C]GHB in mouse brain after an intravenous injection. Cerebral uptake of GHB was rapid and this substance disappeared from brain tissue with a half-life of approx 5 min. Degradation of [1-14C]GHB took place in the brain since 14C was incorporated in amino acids associated with the tricarboxylic acid cycle: the labelling pattern was consistent with the oxidation of GHB via succinate through the cycle, rather than with β-oxidation of GHB. Conversion of [14C]GHB into [14C]GABA prior to oxidation was negligible, thus it is unlikely that the pharmacological action of GHB would be mediated through GABA formation. [14C]GHB oxidation also elicited the signs of metabolic compartmentation of the tricarboxylic acid cycle in the brain (glutamine/glutamate specific radioactivity ratio was about 4).  相似文献   

14.
Abstract: The metabolic fate of glutamate in astrocytes has been controversial since several studies reported >80% of glutamate was metabolized to glutamine; however, other studies have shown that half of the glutamate was metabolized via the tricarboxylic acid (TCA) cycle and half converted to glutamine. Studies were initiated to determine the metabolic fate of increasing concentrations of [U-13C]glutamate in primary cultures of cerebral cortical astrocytes from rat brain. When astrocytes from rat brain were incubated with 0.1 m M [U-13C]glutamate 85% of the 13C metabolized was converted to glutamine. The formation of [1,2,3-13C3]glutamate demonstrated metabolism of the labeled glutamate via the TCA cycle. When astrocytes were incubated with 0.2–0.5 m M glutamate, 13C from glutamate was also incorporated into intracellular aspartate and into lactate that was released into the media. The amount of [13C]lactate was essentially unchanged within the range of 0.2–0.5 m M glutamate, whereas the amount of [13C]aspartate continued to increase in parallel with the increase in glutamate concentration. The amount of glutamate metabolized via the TCA cycle progressively increased from 15.3 to 42.7% as the extracellular glutamate concentration increased from 0.1 to 0.5 m M , suggesting that the concentration of glutamate is a major factor determining the metabolic fate of glutamate in astrocytes. Previous studies using glutamate concentrations from 0.01 to 0.5 m M and astrocytes from both rat and mouse brain are consistent with these findings.  相似文献   

15.
Abstract: Radiolabelled glutamine and glucose were infused into lateral ventricles of rats in order to label transmitter amino acid pools in vivo . Brain regions close to the lateral ventricle (hippocampus, corpus striatum, hypothalamus) were labelled more effectively than more distant structures such as cerebral cortex or cerebellum. All regions were labelled to much the same extent over 30-150 min by [U-14C]glucose, [U-14C]glutamine, or [3H]glutamine administered alone or together in doublelabel experiments when allowance was made for any differences in precursor specific radioactivities. Slices of cerebral cortex or hippocampus from brains labelled in vivo were incubated and stimulated in vitro with veratrine (75 μ M ); tetrodotoxin (1 μ M ) was present in the control medium. Single-label experiments showed that [U-14C]- glutamine was more effective than [U-14C]glucose for labelling releasable glutamate and GABA. Double-label experiments showed that [3H]glutamine and [U-14C]- glucose given together in vivo labelled glutamate and GABA releasable in vitro to a similar extent. Both types of experiment empbasise the large contribution made by glutamine in vivo to pools of transmitter glutamate and GABA.  相似文献   

16.
Abstract: Oligodendroglia prepared from minced calf cerebral white matter by trypsinization at pH 7.4, screening, and isosmotic Percoll (polyvinylpyr-rolidone-coated silica gel) density gradient centrifugation survived in culture on polylysine-coated glass, extending processes and maintaining phenotypic characteristics of oligodendroglia. In the present study, ethanolamine glycerophospholipid (EGP) metabolism of the freshly isolated cells was examined during short-term suspension culture by dual label time course and substrate concentration dependence experiments with [2-3H]glycerol and either [1,2-14C]ethanolamine or L-[U-14C]serine. Rates of incorporation of 3H from the glycerol and of 14C from the ethanolamine into EGP were constant for 14 h. In medium containing 3 mM-[1,2-14C]ethanolamine and 4.8 mM-[2-3H]glycerol, rates of incorporation of 14C and 3H into diacyl glycerophosphoethanolamine (diacyl GPE) were similar. Under the same conditions, 3H specific activities of alkylacyl GPE and alkenylacyl GPE were much lower than 14C specific activities, likely as a result of the loss of tritium during synthesis of these forms of EGP via dihydroxyacetone phosphate. L-[U-14C]serine was incorporated into serine glycerophospholipid (SGP) by base exchange rather than de novo synthesis. 14C from L-[U-14C]serine also appeared in EGP after an initial lag period of several hours. Methylation of oligodendroglial EGP to choline glycerophospholipid (CGP) was not detected.  相似文献   

17.
Abstract— Of seven amino acids studied, glutamic acid and phenylalanine were incorporated in highest amounts into the hot-TCA-insoluble material of the 100,000 g supernatant fraction of rat brain homogenate. The system for incorporation of phenylalanine was RNase-insensitive and required ATP (apparent Km = 0.64 m m ), KC1 (apparent Km = 14 m m ) and MgCl2 (optimal concentration range 4-15 m m ). The apparent Km for phenylalanine was 2.9 m m . [14C]Phenylalanine did not undergo modification before incorporation. Tyrosine and phenylalanine inhibited the incorporation, respectively, of [14C]phenylalanine and [14C]tyrosine when incubated simultaneously or successively. The Km and Kt (3.3 m m ) values for phenylalanine in the incorporation reaction and as inhibitor of the incorporation of [14C]tyrosine were similar. We suggest that both the enzyme and the acceptor for the incorporation of these two amino acids are the same. [14C]Phenylalanine and [14C]tyrosine entered into COOH-terminal positions in the reactions described. Brain exhibited a 25- to 100-fold higher capacity to incorporate phenylalanine than that of liver, kidney or thyroid. The acceptor capacity in rat brain rapidly decreased from day 5 to day 15 of postnatal age and then slowly until age 150 days.  相似文献   

18.
Abstract— Following intracranial and intraperitoneal injection of acetyl- l -[U-14C]aspartate into mice about 5% and 0.7% of the radioactivity, respectively, was recovered from the brain after 30 min.
On chromatographic separation of the cationic and anionic compounds on a Dowex 50 column, the former fraction contained about 60% of the radioactivity, predominantly as labelled aspartate and glutamate. The anionic compounds, containing 20% of the labelled compounds, were fractionated in several chromatographic systems and resolved into a great variety of labelled peptidic compounds of which five acetyl-[U14-C]aspartyl peptides, containing two to four amino acids, were purified. One of these, acetyl-aspartyl glutamine, has not previously been found in brain.  相似文献   

19.
Abstract: The K+-induced release of amino acids and dopamine from synaptosomes of basal ganglia and substantia nigra of sheep was studied. K+ (56 mM) caused an increase in the release of GABA from caudate, putamen, globus pallidus, and substantia nigra, the increased release being 227, 171, 198, and 366%, respectively, compared with samples incubated without stimulation. The release of glutamate was also increased by 56 mM-K+ (136–183%) from all regions except the globus pallidus, and a significant release of aspartate was only seen in response to K+ stimulation of synaptosomes from putamen (50%). Veratrine (75 μM) also stimulated a similar pattern of amino acid release from these regions. Regional correlation was shown between the presence of an uptake system for an amino acid and its evoked release. [14C]Dopamine formed from L-[U-14C]tyrosine was released only from caudate and putamen synaptosomes by K+ stimulation, the increases being 105% and 74%, respectively. Synthesis of [14C]dopamine from L-[U-14C]tyrosine occurred only in synaptosomes prepared from these two regions and was not detected in synaptosomes from substantia nigra or globus pallidus although whole-tissue homogenates of substantia nigra were able to synthesise dopamine.  相似文献   

20.
Abstract: [1-3H, 1-14C]Palmitaldehyde(3H:14C= 15) was injected intracerebrally to 18-day-old rats and incorporation of radioactivity into brain lipids was followed over a 24-h period. The substrate was metabolized primarily by oxidation to palmitic acid with loss of tritium and, to a lesser extent, by reduction to hexadecanol. The alkyl moieties of the ethanolamine phospholipids showed considerably lower 3H:14C ratios than the substrate, indicating a substantial participation in ether lipid synthesis by tritium-free alcohols derived from 14C-labeled fatty acids. Virtually no 3H radioactivity was found in alkenyl moieties, indicating stereospecificity of both reduction of aldehyde and dehydrogenation of alkyl to alkenyl glycerolipid. The data are consistent with the general concept that plasmalogen biosynthesis proceeds exclusively through fatty alcohols and alkyl glycerolipids and that fatty aldehydes cannot be utilized directly.  相似文献   

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