Trp Pyrrolase Research Articles

Activation of Liver Tryptophan Pyrrolase Mediates the Decrease in Tryptophan Availability to the Brain after Acute Alcohol Consumption by Normal Subjects

We have previously suggested that acute ethanol consumption by normal subjects decreases the availability of circulating tryptophan (Trp) to the brain by activating liver Trp pyrrolase, the first and rate-limiting enzyme of the (major) kynurenine pathway of Trp degradation. The aim of the present study was to examine this hypothesis further by measuring plasma levels of kynurenine metabolites following alcohol consumption. After an overnight fast and a light breakfast, each of 10 healthy subjects received one of five drinks (placebo and doses of ethanol of 0.2, 0.4, 0.6 and 0.8 g/kg body weight in tonic water) on five different occasions. Blood samples were withdrawn 2 h later and plasma was analysed for concentrations Trp, competing amino acids (CAA) and kynurenine metabolites. Along with the depletion of plasma Trp and the decrease in its availability to the brain, as expressed by the ratio of [Trp]/[CAA], plasma kynurenine was elevated by doses of ethanol of 0.2-0.8 g/kg body weight. The ratio% of [kynurenine]/[Trp], an index of the expression of Trp pyrrolase activity, was also increased by all doses of ethanol. We conclude that activation of liver Trp pyrrolase mediates the depletion of plasma Trp and the decrease in its availability to the brain induced by acute ethanol consumption.

Tryptophan metabolism in alcoholism

Acute and chronic alcohol (ethanol) intake and subsequent withdrawal exert major effects on tryptophan (Trp) metabolism and disposition in human subjects and experimental animals. In rats, activity of the rate-limiting enzyme of Trp degradation, liver Trp pyrrolase (TP), is enhanced by acute, but inhibited after chronic, ethanol administration, then enhanced during withdrawal. These changes lead to alterations in brain serotonin synthesis and turnover mediated by corresponding changes in circulating Trp availability to the brain. A low brain-serotonin concentration characterizes the alcohol-preferring C57BL/6J mouse strain and many alcohol-preferring rat lines. In this mouse strain, liver TP enhancement causes the serotonin decrease. In man, acute ethanol intake inhibits brain serotonin synthesis by activating liver TP. This may explain alcohol-induced depression, aggression and loss of control in susceptible individuals. Chronic alcohol intake in dependent subjects may be associated with liver TP inhibition and a consequent enhancement of brain serotonin synthesis, whereas subsequent withdrawal may induce the opposite effects. The excitotoxic Trp metabolite quinolinate may play a role in the behavioural disturbances of the alcohol-withdrawal syndrome. Some abstinent alcoholics may have a central serotonin deficiency, which they correct by liver TP inhibition through drinking. Further studies of the Trp and serotonin metabolic status in long-term abstinence in general and in relation to personality characteristics, alcoholism typology and genetic factors in particular may yield important information which should facilitate the development of more effective screening, and preventative and therapeutic strategies in this area of mental health.

Rat liver tryptophan pyrrolase activity and gene expression during alcohol withdrawal.

Rat liver tryptophan (Trp) pyrrolase activity and gene expression were studied in relation to the alcohol-withdrawal syndrome (AWS). Both activity and gene expression were enhanced after withdrawal of ethanol-containing liquid diets and the time-course of these changes mirrored that of development and intensity of the behavioural disturbances of the AWS. By contrast, no correlation was observed between the AWS-induced behaviour and changes in activity of another hepatic glucocorticoid-inducible enzyme, tyrosine aminotransferase, and a negative correlation was noted between behaviour and the gene expression of this latter enzyme and also of that of the hepatic glucocorticoid receptor. We suggest that the metabolic consequences of activation of liver Trp pyrrolase during alcohol withdrawal may play a role in the behavioural features of the AWS.

Tryptophan metabolism in alcoholism.

Studies of tryptophan (Trp) metabolism in relation to the serotonin status in alcoholism are of 2 types: (1) those related to the pharmacological effects of ethanol; (2) those concerning the serotonin status in the absence of alcohol intake. In experimental animals, acute and chronic ethanol administration and subsequent withdrawal exert a variety of effects on brain serotonin synthesis and turnover mediated by corresponding changes in Trp availability to the brain secondarily mainly to modulation of liver Trp pyrrolase (TP) activity. Alcohol-preferring mice and rats exhibit a central serotonin deficiency caused by, or in some cases associated with, a higher TP activity. Liver TP also appears to be a target of ethanol in man and evidence has recently emerged that alcoholics with positive family history are serotonin-deficient because of a lower availability of circulating Trp to the brain. Acutely, ethanol depletes brain serotonin in normal subjects, which may explain alcohol-induced aggression in susceptible individuals and also the incidence of depression in alcoholism. Trp availability to the brain is increased before the appearance of the alcohol-withdrawal syndrome in man, raising the possibility that the associated behavioural disturbances may involve the excitotoxic Trp metabolite quinolinate. Further studies of the Trp and serotonin status in relation to these important clinical features of alcohol dependence and alcoholism may therefore yield fruitful results.

Tryptophan metabolism in male Sardinian alcohol-preferring (sP) and -non-preferring (sNP) rats.

Parameters of tryptophan (Trp) and related metabolism were compared in male Sardinian alcohol-preferring (sP) and -non-preferring (sNP) rats. Liver Trp pyrrolase activity was 38-58% higher in sP than in sNP rats, and this was associated with a greater expression of the enzyme mRNA as measured by multiprobe oligonucleotide solution hybridization. Moderately (about 10-19%), but significantly, lower concentrations of free serum, total serum, and brain Trp were also observed in sP compared with sNP rats. Concentrations of whole brain 5-hydroxytryptamine (5-HT) and its major metabolite 5-hydroxyindol-3-yl-acetic acid (5-HIAA) were, however, 14-21% higher in sP rats. Serum corticosterone concentration was 18% higher in sP rats. We conclude that alcohol preference in Sardinian rats is associated with increased liver Trp pyrrolase activity and mRNA expression leading to a decrease in Trp availability to the brain. Although a simple serotonin deficiency could not be demonstrated in the whole brain, the possibility could not be ruled out that a deficiency may be present in discrete areas of the brain of the sP rat.

Effects of chronic administration and subsequent withdrawal of ethanol-containing liquid diet on rat liver tryptophan pyrrolase and tryptophan metabolism.

An investigation of the effects of chronic administration of ethanol by the liquid diet procedure and its subsequent withdrawal on tryptophan (Trp) metabolism and disposition was performed in rats. Treatment with the control liquid diet caused an enhancement of liver Trp pyrrolase activity and mRNA abundance. These effects are not due to the starvation associated with this feeding procedure, because they occur in rats maintained on the liquid diet ad libitum. Chronic ethanol administration in the liquid diet did not further influence the above increased expression of Trp pyrrolase mRNA but caused inhibition of pyrrolase activity in competition with the effects of the diet. The control liquid diet decreased liver Trp concentration, but exerted no significant effects on other aspects of Trp disposition. The most striking and robust finding was a highly significant elevation in both Trp pyrrolase activity and mRNA expression at 7 h following discontinuation of ethanol availability, at which time there were demonstrable behavioural signs of ethanol withdrawal. The increase in Trp pyrrolase mRNA during alcohol withdrawal may be caused by corticosterone, whose circulating concentration was also increased. The changes in Trp pyrrolase activity during ethanol withdrawal were associated with significant alterations in Trp disposition including decreased brain Trp concentration and 5-hydroxytryptamine synthesis and turnover. These alterations may play a pivotal role in the behavioural manifestations of ethanol withdrawal including the hyperexcitement underlying audiogenic seizures. We suggest that rat Trp pyrrolase gene regulation may be an important biological determinant of the ethanol withdrawal syndrome and requires further study, and that the use of the liquid diet procedure in Trp metabolic studies requires inclusion of adequate controls and special attention to the effects of the liquid diet itself.

Mechanism of Enhancement of Rat Brain Serotonin Synthesis by Acute Fluoxetine Administration

Tsuiki et a!. (1995) reported in this Journal that acute intraperitoneal administration of a 30 mg/kg of body weight dose of the specific serotonin [5-hydroxytryptamine (5-HT)11 reuptake inhibitory (SSRI) antidepressant fluoxetine enhances rat brain 5-HT synthesis, as measured by the a-methyl-L-tryptophan autoradiographic method, and suggested that this is in part due to an increase in tryptophan (Trp) availability to the brain. This process is determined by at least three peripheral factors: binding to albumin, competition between Trp and five other amino acids (valine, leucine, isoleucine, phenylalanine, and tyrosine) for the same cerebral uptake mechanism, and activity of the major Trp-degrading enzyme, hepatic Trp pyrrolase (tryptophan 2,3-dioxygenase; EC 1.13.11.11) (for references, see Badawy and Morgan, 1991). Tsuiki et al. (1995) showed that fluoxetine does not influence the concentrations of the Trp competitors, but increases that of Trp itself (by 18% for the total content and 30% for the free fraction). Whether Trp binding to albumin was significantly altered by fluoxetine could not be ascertained from the data given.

Tryptophan metabolism and disposition in relation to alcohol and alcoholism.

Tryptophan (Trp) metabolism and disposition in relation to alcohol and alcoholism are briefly reviewed. The changes observed could generally be classified into those: (1) exerted by acute or chronic alcohol administration and/or subsequent withdrawal; (2) already present in the absence of alcohol consumption, such as in naive alcohol-preferring animals or in abstinent alcoholics. In normal rats, acute ethanol administration activates liver Trp pyrrolase and exerts a biphasic effect on brain 5-HT (5-hydroxytryptamine or serotonin) synthesis, whereas chronic ethanol administration and subsequent withdrawal exert opposite effects on 5-HT synthesis mediated by corresponding changes in liver Trp pyrrolase activity. A cerebral 5-HT deficiency has been demonstrated in the alcohol-preferring C57BL mouse strain and in a number of alcohol-preferring rat lines, the mechanism of which is understood only in two models; the C57B1 mouse strain has a higher liver Trp pyrrolase activity and the P rat line from Indiana has a lower density of serotonergic fibres in cerebral cortex. In man, acute ethanol intake lowers circulating [Trp] and its availability to the brain, almost certainly by activating liver Trp pyrrolase. Some evidence exists for possible inhibition of pyrrolase activity in non-abstinent chronic alcoholics. Evidence in recently abstinent alcoholics suggests that Trp availability to the brain may be impaired and that this may be particularly so in patients with positive family history. Exploration of this latter possibility may be important in understanding the biological basis of predisposition to alcoholism.

Decrease in Circulating Tryptophan Availability to the Brain After Acute Ethanol Consumption by Normal Volunteers: Implications for Alcohol-Induced Aggressive Behaviour and Depression

Acute ethanol consumption by fasting male volunteers decreases circulating trytophan (Trp) concentration and availability to the brain as determined by the ratio of (Trp) to the sum of its five competitors ([Trp]/[CAA]ratio). These effects of alcohol are specific to Trp, because levels of the 5 competitors are not increased. The decrease in circulating (Trp) is not associated with altered binding to albumin and may therefore be due to enhancement of hepatic Trp pyrrolase activity. It is suggested that, under these conditions brain serotonin synthesis is likely to be impaired and that, as a consequence, a possible strong depletion of brain serotonin in susceptible individuals may induce aggressive behaviour after alcohol consumption. The possible implications of these findings in the relationship between alcohol and depression are also briefly discussed.

Dietary requirements of rainbow trout for tryptophan, lysine and arginine determined by growth and biochemical measurements

Three separate studies were performed to determine the dietary requirements of rainbow troutSalmo gairdneri for tryptophan (Trp), lysine (Lys) and arginine (Arg) from both growth and biochemical data. The growth studies were carried out over a 12 week period. From graphical plots of % mean weight gain against % amino acid in diet the following requirement values were obtained, Trp 0.25% diet (0.4% dietary crude protein); Lys 1.9% diet (4.3% dietary protein); and Arg 1.6-1.8% diet (3.6-4% dietary protein). Plasma and liver amino acid concentrations measured 20h after feeding did not prove useful for determination of requirement values. Hepatic activities of Trp pyrrolase (TP), Lys α ketoglutarate reductase (LKGR) and arginase were not significantly affected by varying levels of Trp, Lys and Arg respectively in the diet. TP has a cytosolic location and a Km of 0.2 mM for Trp; LKGR is mitochondrial and the Km for Lys is 7.3 mM; arginase is also mitochondrial and has a Km of 4.9 mM for arginine. Measurements of expired(14)CO2, after injection of a tracer dose of(14)C amino acid, did allow estimates of requirement levels to be made. The values obtained from the oxidation studies reinforced the values obtained from the growth data but were not precise enough to justify using this method on its own.

Effects of combined administration of DL-3-pyridylalanine and 5-hydroxy-L-tryptophan on brain and peripheral tissue serotonin contents in rats

1) Repeated administration of 5-hydroxy-L-tryptophan (5-HTP, 5, 10 or 50 mg/kg/d, 7d) did not produce any further increase in brain serotonin (5-HT) content in Wistar male rats as compared with that found in animals given a single dose of 5-HTP. On the other hand, 5-HT content in peripheral tissues (liver, kidney, spleen and small intestine) increased more after the repeated administration of 5-HTP than after a single dose of this compound. The accumulation of 5-HT in these tissues increased with an increase in the dosage and the number of administration times of 5-HTP. 2) Brain 5-HT content increased more and maintained a high level by the combined administration of DL-3-pyridylalanine (3-PA, 100 mg/kg) with 5-HTP (5 or 10 mg/kg) than by the single administration of these compounds ; the effect was additive. Liver, kidney, spleen and small intestine 5-HT contents after the combined administration of 3-PA and 5-HTP were not different from those after the single administration of 5-HTP. 3) The efficacy of combined administration of 3-PA and 5-HTP was further confirmed by the results of the determination of tryptophan (Trp)-5-HT metabolizing enzyme activities (Trp pyrrolase, Trp 5-hydroxylase, 5-HTP decarboxylase and monoamine oxidase) and pharmacological experiment (conditioned avoidance response) in rats given these compounds. 4) Data are discussed in relation to the side effect of 5-HTP for the treatment of depression.