Rat Brain Tryptophan Research Articles

Self-Selected Meal Composition, Circadian Rhythms and Meal Responses in Plasma and Brain Tryptophan and 5-Hydroxytryptamine in Rats

Relationships among food composition, plasma and brain tryptophan and 5-hydroxytryptamine (5-HT) were studied by measuring both circadian feeding and meal responses in self-selecting rats. Under ad libitum feeding conditions in which young rats were given a choice between two isocaloric diets (0 and 60% casein) and groups were killed at 3-hour intervals throughout the day, several circadian rhythms become apparent. Protein and carbohydrate energy, the plasma tryptophan:neutral amino acid ratio (TRP:NAA) and brain TRP, but not 5-HT concentrations showed statistically significant circadian variations. During the dark hours the plasma TRP:NAA ratio and brain TRP concentrations were inversely associated with the protein concentration of meals previously selected. In meal response studies, rats were similarly given a choice of diets, but were killed 20 minutes after their selected meals. Plasma TRP:NAA ratios, brain TRP and 5-hydroxyindole levels were inversely correlated with the protein concentration of the meals selected. Absolute quantities of protein, but not carbohydrate, fat or total energy, were inversely related to the plasma TRP:NAA ratio and brain TRP concentration. It is concluded that the circadian changes in the plasma TRP:NAA ratio and in brain TRP and 5-hydroxyindole concentrations are under the continual influences of food choices made during spontaneous feeding activity.food selection food intake regulation circadian rhythms 5-hydroxytryptamine tryptophan protein concentration

An Improved High Performance Liquid Chromatographic Method for Tryptophan Analysis in Rat Brain Administrated by Seaweed

IBrain functions can be affected by the dietary precursors of neurotransmitters. Tryptophan (TRP) is the precursor of serotonin (5- Hydroxytryptamine; 5-HT) neurotransmitter that plays an important role in sleeping, mood, eating behavior, consciousness and cognitive functions. Present study describes the estimation of tryptophan in rat brain by aqueous-reverse-phase high performance liquid chromatography. Estimation was carried out on a shim pack end-capped C-18 column using water as mobile phase (pH=9 adjusted by addition of 0.1 mM NaOH) at a flow rate of 1 ml min-1. Eluents were monitored at 273 nm by an ultra-violet detector. The method was linear (R2=0.9973) and the limit of detection (LOD) and limit of quantification (LOQ) were 4.3920 and 13.3091 ng ml-1, respectively. The developed HPLC method is simple convenient, reliable and economical for the estimation of tryptophan in rat brain. In the present study, Albino Wistar male rats were randomly divided into two groups control and seaweeds treated rats. Seaweeds (Jolyna Laminiriodes) were orally given to test rats at a dose of 200 mg kg-1 whereas control rats were given vehicle for 28 days. After 28 days of treatment, rats were decapitated and its brain samples were taken out for the estimation of TRP. Current study observed increased in TRP levels in the brain samples of seaweed treated rats as compared to control rats. Therefore present study suggested the use of seaweeds in the treatment of disorders caused by TRP depletion. By this newly developed method for the estimation of TRP, effect of various drugs on TRP levels in the brain could be studied.

Rapid analysis of neurotransmitters in rat brain using ultra‐fast liquid chromatography and tandem mass spectrometry: application to a comparative study in normal and insomnic rats

Neurotransmitters and their metabolites in central nervous system were known to play a significant role in sedation and hypnosis. A rapid and sensitive UFLC-MS/MS method for simultaneous determination of serotonin, 5-hydroxyindole acetic acid (5-HIAA), tryptophan (Try), dopamine (DA), norepinephrine (NE), γ-aminobutyric acid (GABA), glutamic acid (Glu) and acetylcholine (Ach) in rat brain without derivatization, ion-pairing reagent or pre-concentration was developed. Analytes and IS were separated on a Inertsil ODS-EP column (150 mm × 4.6 mm, 5 µm particles) and analyzed in a single chromatographic run in less than 9.0 min, using gradient elution with the mobile phase consisting of methanol and 0.01% acetic acid in water at a flow rate of 1.2 ml min(-1) . The detection of the analytes was performed on 4000Q UFLC-MS/MS system with turbo ion spray source in positive ion and multiple reaction monitoring mode. The developed method provided excellent linear calibration curves for the assay of analytes (R(2) ≥ 0.9915). Limits of quantification were in the range of 1.0 ng ml(-1) to 1.0 µg ml(-1) for the analytes in rat brain. Intra- and inter-day precision and accuracy of analytes were well within acceptance criteria (15%). Mean extraction recoveries of analytes and IS from rat brain were all more than 80.0%. Furthermore, the validated method was successfully applied to comparing profiles of analytes in normal and insomnic rat brains. Results indicated that there were statistically significant differences for serotonin, 5-HIAA, DA, NE, Glu and Ach, but no significant difference for Try and GABA between two groups.

Ключові показники шляху біосинтезу серотоніну в мозку щурів за умов хронічної дії алкоголю

Key indicators of serotonin synthesis in the brain of rats with experimental chronic alcohol intoxication were investigated. Content of tryptophan in rat brain during the period of chronic alcohol intoxication was increased. During withdrawal symptoms, a reduction in tryptophan content in comparison with its value during the administration of alcohol solution was shown. An increased activity, a key and rate limiting enzyme of serotonin biosynthesis path – tryptophan-hydroxylase in rat brain at later stages of experimental chronic alcohol intoxication was shown. Serotonin content in rat brain decreased during the period of chronic alcohol intoxication. At the 11th day of administration of 30% solution of ethanol and during the withdrawal syndrome, growth of monoaminoxygenase activity – enzyme that controls the degradation of 5-hydroxytryptamine – was shown. These results suggest the involvement of serotonergic neurotransmitter systems in pathogenesis of brain development at chronic alcohol intoxication in rats. Keywords: serotonin, tryptophan, chronic alcohol intoxication.

Determination of Tryptophan in Raw Materials, Rat Brain and Human Plasma by RP-HPLC Technique

This paper describes tryptophan (TRP) estimation in raw human plasma and rat brain by reversed-phase high-performance liquid chromatography (RP-HPLC). Estimation was carried out on a Purospher STAR C18 column using water-acetonitrile (90:10 v/v, at pH 2.7) mixture at a rate of 1.5 mL/min as mobile phase. Eluents were monitored at 273 nm by an ultraviolet detector. The method was linear (R(2) > 0.999), precise (intra-day and inter-day precision <2%) in the range of 0.25-20 µg/mL. The detection and quantification limits were 0.0144 µg/mL and 0.0437 µg/mL, respectively. In human plasma, Day 1 and Day 2 precision were 0.054-2.29% and 1.66-3.7%; whereas precisions in rat brain were 1.23-2.3% and 0.677-4.2%, respectively. The method was applied to study TRP level in human smokers and in arthritic rat brain. An efficient RP-HPLC method was developed for TRP determination that worked for clinical and research purposes.

Effect of ethanol on the hydroxylation of tyrosine and tryptophan in rat brain in vivo.

Abstract Various doses of ethanol were injected intraperitoneally into rats. After 20 min, 3-hydroxybenzylhydrazine (NSD 1015), an inhibitor of aromatic amino-acid decarboxylase was injected and 30 min later the animals were killed. The amount of dopa accumulating in brain as a consequence of decarboxylase inhibition was significantly increased by moderate doses of ethanol (up to 4 g kg−1). There was no corresponding effect on 5-hydroxytryptophan. The effect on dopa was found both in a dopamine- and a noradrenaline-dominated area of the brain, suggesting that catecholamine synthesis is stimulated by ethanol in both types of neuron. The data lend support to the hypothesis of Carlsson, Engel &amp; Svensson (1972) that cerebral catecholamines are involved in the central stimulation induced by ethanol.

Amitriptyline and clomipramine increase the concentration of administered L-tryptophan in the rat brain.

The tricyclic antidepressant amitriptyline has been shown to reduce concentrations of large neutral amino acids (LNAA) in rat plasma. Compounds with that property might interact with such amino acids used as therapeutic agents with a central site of action by causing a change in the relationship between the administered LNAA and its endogenous LNAA competitors for carrier-mediated transport through the blood-brain barrier into the brain. This study was performed to investigate if the antidepressant agents amitriptyline and clomipramine could, by such a mechanism, increase brain concentrations of administered tryptophan. Intraperitoneal administration of L-tryptophan alone (100 mg kg-1) resulted in an increase in the concentration of tryptophan in the rat brain from 14 +/- 0.7 to 100 +/- 4.3 nmol g-1 compared with rats given saline only. When rats were given tryptophan with amitriptyline (25 mg kg-1, i.p.) or clomipramine (25 mg kg-1, i.p.) brain concentrations of tryptophan were increased even further, to 150 +/- 4.5 and 157 +/- 10.2 nmol g-1, respectively. Administration of L-tryptophan alone resulted in an increase in the rat plasma tryptophan ratio [(concentration of tryptophan)/(total concentration of LNAAs)] from 0.14 +/- 0.003 to 0.42 +/- 0.011 compared with rats given saline only. When rats were given tryptophan with amitriptyline or clomipramine the plasma tryptophan ratios were increased even further to 0.52 +/- 0.017 and 0.54 +/- 0.025, respectively. All these effects were statistically significant (P < 0.001). These findings support the hypothesis that tricyclic antidepressants could interact with administered tryptophan by changing the relationship in plasma between tryptophan and its endogenous LNAA competitors for transport into the brain, resulting in higher concentrations of tryptophan in the brain. It is possible that this could be the mechanism of the previously reported finding that clomipramine and tryptophan potentiate each other in the treatment of depression.

Chronic melatonin treatment and its precursor L-tryptophan improve the monoaminergic neurotransmission and related behavior in the aged rat brain

Melatonin has an important role in the aging process as a potential drug to relieve oxidative damage, a likely cause of age-associated brain dysfunction. As age advances, the nocturnal production of melatonin decreases potentially causing physiological alterations. The present experiments were performed to study in vivo the effects of exogenously administered melatonin chronically on monoaminergic central neurotransmitters serotonin (5-HT), dopamine (DA) and norepinephrine (NE) and behavioral tests in old rats. The accumulation of 5-hydroxy-tryptophan (5-HTP) and L-3,4-dihydroxyphenylalanine (DOPA) after decarboxylase inhibition was used as a measure of the rate of tryptophan and tyrosine hydroxylation in rat brain. Also neurotransmitters 5-HT, DA and NE and some metabolites were quantified by HPLC. In control rats, an age-related decline was observed in neurochemical parameters. However, chronic administration of melatonin (1 mg/kg/day, diluted in drinking water, 4 wk) significantly reversed the age-induced deficits in all the monoaminergic neurotransmitters studied. Also, neurochemical parameters were analyzed after administration of melatonin biosynthesis precursor L-tryptophan (240 mg/kg/day, i.p., at night for 4 wk) revealing similar improvement effects to those induced by melatonin. Behavioral data corresponded well with the neurochemical findings since spatial memory test in radial-maze and motor coordination in rota-rod were significantly improved after chronic melatonin treatment. In conclusion, these in vivo findings suggest that melatonin and L-tryptophan treatments exert a long-term effect on the 5-HT, DA and NE neurotransmission by enhancing monoamine synthesis in aged rats, which might improve the age-dependent deficits in cognition and motor coordination.

Effect of reactive oxygen species on the metabolism of tryptophan in rat brain: influence of age.

In an earlier study, oxidation of tryptophan hydroxylase was implicated as its affinity was decreased with aging in rat brain. To establish any potential link between its oxidative damage and aging, we have determined the activities of antioxidant enzymes in midbrain, pons and medulla of 2, 12 and 24 month old Fisher 344 BNF1 rats. The results obtained suggest that the activities of antioxidant enzymes varied considerably with age and brain regions studied. Activities of Cu/Zn superoxide dismutase and glutathione peroxidase were found to increase from 2 to 12 months and then decrease in 24 month old rats. However catalase activity decreased consistently with the age. A parallel increase in the carbonyl content was observed in these brain regions indicating the oxidation of proteins. Reactive oxygen species when included in the incubation mixture decreased the activity of tryptophan hydroxylase in a concentration dependent manner. The loss of tryptophan hydroxylase activity induced by hydrogen peroxide and superoxide anion was prevented by catalase. However superoxide dismutase did not provide such protection. Sulfhydryl agents, cysteine, glutathione and dithiothreitol partially prevented the loss of activity. These studies suggest an involvement of reactive oxygen species for sulfhydryl oxidation of tryptophan hydroxylase in aging.

Influence of tryptophan supplementation of soy-based infant formulas on protein quality and on blood and brain tryptophan and brain serotonin in the rat model.

A 2-wk feeding study with weanling rats was conducted to investigate the effects of tryptophan supplementation of soy-based infant formulas on protein quality and on the levels of blood and brain tryptophan and brain serotonin and 5-HIAA (5-hydroxyindole-3-acetic acid), a metabolite of serotonin. The powder and liquid concentrate forms of soy-based infant formulas obtained from four manufacturers were tested. The tryptophan contents of the test soy-based formulas (10.0-12.3 mg/g protein) were considerably lower compared to those of human milk (17-19 mg/g protein). The infant formulas were fed as the sole source of protein in diets containing 8% protein, 20% fat, 5% cellulose and adequate amounts of minerals and vitamins. In general, supplementation of the diets containing test formulas with graded levels of L-tryptophan (0.1 or 0.2%) had no effect on protein quality indices based on rat growth such as PER (protein efficiency ratio), NPR (net protein ratio) and relative NPR (RNPR), but resulted in significant (p < 0.05) increases in the concentrations of tryptophan in the plasma and brain, and serotonin and 5-HIAA in brain of rats. The increased concentrations of brain tryptophan and 5-hydroxyindoles (serotonin + 5-HIAA) in rats fed tryptophan-supplemented formulas compared to those fed unsupplemented formulas support the need for further research to investigate the influence of tryptophan supplementation of soy-based infant formulas on tryptophan metabolites and their potential related effects on sleep latency and neurobehavioral developments in infants.

Lipopolysaccharide administration produces time-dependent and region-specific alterations in tryptophan and tyrosine hydroxylase activities in rat brain.

The present study examined the effect of systemic administration of lipopolysaccharide (LPS; 100 and 250 microg/kg, i.p.) on tyrosine hydroxylase (TH) and tryptophan hydroxylase (TPH) activities in frontal cortex, striatum and midbrain of the rat. Enzyme activities were determined by measuring accumulation of the transient intermediates 5-hydroxytrptophan (5-HTP) and L-dihydroxyphenylalanine (L-DOPA) following in vivo administration of the decarboxylase inhibitor, NSD 1015. TPH activity was increased 2 hours after administration of LPS (100 and 250 microg/kg) in both frontal cortex and midbrain, and a secondary increase was seen in the midbrain 12 hours after challenge. LPS provoked an increase in TH activity in the midbrain only, and this was evident for up to 24 hours after LPS administration. Thus in addition to previous studies demonstrating that LPS increases in vivo NA, DA and 5-HT release, this study shows that LPS increases the activity of the rate-limiting enzymes responsible for their synthesis.

Liquid Concentrates Are Lower in Bioavailable Tryptophan than Powdered Infant Formulas, and Tryptophan Supplementation of Formulas Increases Brain Tryptophan and Serotonin in Rats

The bioavailability of tryptophan in powdered and/or liquid concentrate forms of milk-based infant formulas was determined by studying rat growth response by using a slope ratio method (food conversion efficiency: weight gain/food consumed vs. tryptophan consumed). A gelatin basal diet formulated to be adequate in all nutrients, except tryptophan (0.03%), for rat growth was supplemented with graded levels of crystalline L-tryptophan (0.02, 0.04, 0.06, 0.08, 0.10, 0.12 and 0.14%, standard diets) or infant formulas providing 0.04 and 0.08% supplemental tryptophan (test diets). These diets were fed to weanling rats for 2 wk. Tryptophan bioavailabilities of various formulas varied from 83 to 95%, with some of the liquid concentrates having the lowest values. The levels of bioavailable tryptophan in the liquid concentrate forms (9.7–12.6 mg/g protein) and the powdered forms (11.1–13.1 mg/g protein) were considerably lower than those of human milk (17–19 mg/g protein). Supplementation of the liquid concentrates with graded levels of L-tryptophan (0.1, 0.5 and 1.0%) had no effect on protein quality indices, based on rat growth, but resulted in a dose-related increase in the concentrations of tryptophan in the plasma and brain and of serotonin and 5-hydroxyindole-3-acetic acid in the brains of rats. This study supports further research to investigate the influence of tryptophan supplementation of infant formulas, to more closely simulate tryptophan composition of human milk, on tryptophan metabolites and their potential related effects on sleep latency and neurobehavioral developments in infants.

24h withdrawal following repeated administration of caffeine attenuates brain serotonin but not tryptophan in rat brain: Implications for caffeine-induced depression

Caffeine injected at doses of 20, 40 and 80 mg/kg increased brain levels of tryptophan, 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) in rat brain. In view of a possible role of 5-HT in caffeine-induced depression the effects of repeated administration of high doses of caffeine on brain 5-HT metabolism are investigated in rats. Caffeine was injected at doses of 80 mg/kg daily for five days. Control animals were injected with sahne daily for five days. On the 6th day caffeine (80 mg/kg) injected to 5 day sahne injected rats increased brain levels of tryptophan, 5-HT and 5-HIAA. Plasma total tryptophan levels were not affected and free tryptophan increased. Brain levels of 5-HT and 5-HIAA but not tryptophan decreased in 5 day caffeine injected rats injected with sahne on the 6th day. Plasma total and free tryptophan were not altered hi these rats. Caffeine-induced increases of brain tryptophan but not 5-HT and 5-HIAA were greater in 5 day caffeine than 5 day sahne injected rats. The findings are discussed as repeated caffeine administration producing adaptive changes in the serotonergic neurons to decrease the conversion of tryptophan to 5-HT and this may precipitate depression particularly in conditions of caffeine withdrawal.

Early induction of rat brain tryptophan hydroxylase (TPH) mRNA following parachlorophenylalanine (PCPA) treatment

Tryptophan hydroxylase (TPH) is the first and presumably rate-limiting enzyme in serotonin (5-HT) biosynthesis. End-product inhibition of rate-limiting enzymes is common and 5-HT is known to inhibit TPH activity in vivo. However, it is not known whether levels of 5-HT could also be involved in the regulation of the TPH gene. In order to determine whether TPH gene regulation is dependent on the 5-HT concentration, 5-HT levels were reduced by the administration of parachlorophenylalanine (PCPA). PCPA is a potent, specific and irreversible inhibitor of TPH activity which drastically reduces 5-HT concentration in the 5-HT neurons and terminals. When PCPA was administered, TPH activity in both cell bodies and nerve terminal areas, was reduced to 10% of control values and recovered to the control levels by day 7 in raphe nucleus, and within 14 days in the hypothalamus. In serotonergic terminal areas, 5-HT could not be detected immunohistochemically at day 1, but slowly recovered within 2 weeks. At all time points examined, aromatic l-amino acid decarboxylase (AADC) levels were not changed either in the cell body or terminal areas. The steady state levels of TPH mRNA estimated by in situ hybridization increased at day 1 and returned to control levels by day 4. AADC message levels were not altered throughout the periods. These data suggest that a decrease in 5-HT concentration may lead to an up-regulation of TPH gene transcription, by an, as yet, unknown mechanism. Taken together, our results suggest that feedback control of the rate-limiting enzyme, TPH, may occur not only through changes in enzyme activity, but also through changes in gene transcription (Preliminary results were published by Park et al. [ Soc. Neurosci. Abstr., 17 (1991) 1176] at the Neuroscience Meeting in 1991).

Radioenzymatic assay for tryptophan hydroxylase: [ 3H]H 2O release assessed by charcoal adsorption

The rate-limiting reaction in the biosynthesis of the neurotransmitter, serotonin, is catalyzed by the enzyme, tryptophan hydroxylase. Studies on the characteristics of this enzyme have been hampered by its relative instability and paucity in the brain. We have modified a charcoal adsorption radioenzymatic assay used for the measurement of tyrosine hydroxylase to assess rat brain tryptophan hydroxylase activity. This protocol is based on the principle that aromatic amino acid hydroxylases are mixed-function oxygenases and will utilize O2 and reduced pterin to convert tritiated amino acid substrate to product and tritiated H2O. All product and unreacted substrate are adsorbed by acidified charcoal. The [3H]H2O is analyzed by liquid scintillation spectrometry and is indicative (stoichiometrically) of the amount of product formed and, thus, the activity of the enzyme. This assay has a high signal-to-noise ratio and is sensitive enough to determine enzymatic activity in homogenates of individual raphe nuclei. In addition, its simplicity in design allows for the simultaneous testing of large numbers of samples. The enzyme activity and kinetic determinations derived from this protocol agree with those of other investigators using more lengthy, involved procedures.

Accumulation of Brain Tryptophan in Rats after Administering Various Fats or Fatty Acids.

The intragastric administration of various kinds of fat (corn oil, lard, safflower oil, perilla oil, and fish oil) or fatty acids (oleic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA)) caused a significant increase in brain tryptophan (especially with fish oil or DHA), and also in serum insulin. However, at 2 h after administering the fats or fatty acids, brain serotonin was not increased in accordance with the increase of brain tryptophan. As a result of determining the time-dependent changes in brain 5-hydroxyindoles, brain serotonin and 5-HIAA gradually increased, and showed significant increases at 6 hand 4 h after the administration of fish oil, respectively. These results suggest that there was a fairly long time-lag for the changes in brain 5-hydroxyindoles to appear after being induced by the administration of fats or fatty acids.

Restriction of Maternal Dietary Carbohydrate Decreases Fetal Brain Indoles and Glycogen in Rats

In spite of evidence that dietary carbohydrate can increase brain tryptophan and 5-hydroxytryptamine in adult rats, the possible influence of maternal dietary carbohydrate on fetal brain indoles has received little attention. We studied the effect of graded levels (0, 4, 12 and 60%) of maternal dietary fructose or glucose fed throughout pregnancy on fetal brain glycogen and indoles. The diets were isoenergetic and met the NRC energy requirements for pregnant rats. The results demonstrated that low maternal dietary carbohydrate, with adequate energy intake, reduced fetal brain weight and concentrations of glycogen, tryptophan, 5-hydroxytryptamine and 5-hydroxyindoleacetic acid. There were no significant differences between glucose and fructose feeding at any dietary carbohydrate level for any fetal brain measurements, showing that it was the level and not the type of dietary carbohydrate that was important. Significant correlations between fetal brain 5-hydroxytryptamine and brain glycogen, and between fetal brain 5-hydroxytryptamine and brain weight, suggested that lowered brain 5-hydroxytryptamine was only one symptom of disrupted brain development in fetuses of dams fed low levels of carbohydrate. The results show that dietary carbohydrate restriction during pregnancy can have adverse effects on fetal brain development, glycogen levels, and neurotransmitter synthesis even when maternal dietary protein and energy intake are adequate.

"7-tetrahydrobiopterin," a naturally occurring analogue of tetrahydrobiopterin, is a cofactor for and a potential inhibitor of the aromatic amino acid hydroxylases.

The ability of 2-amino-4-hydroxy-7-[dihydroxylpropyl-(L-erythro)-5,6,7,8-tetrahyd ropterin] ("7-tetrahydrobiopterin" or 7-BH4) to substitute for the natural cofactor tetrahydrobiopterin (BH4) has been studied in vitro in the reactions of the three mammalian aromatic amino acid hydroxylases. With rat liver phenylalanine hydroxylase, the apparent Km for 7-BH4 is 160 microM, a value that is approximately 60-fold greater than that for the natural cofactor. In contrast, the hydroxylase reaction is severely inhibited by as little as 1 microM 7-BH4 when assayed in the presence of physiological concentrations of BH4. This inhibition can be overcome either by an increase in the concentration of BH4 or a decrease in the concentration of phenylalanine. With both rat brain tryptophan hydroxylase and rat pheochromocytoma tyrosine hydroxylase, the Km value for 7-BH4 is about one order of magnitude greater than the Km for BH4. Accordingly, 7-BH4 is a poor competitive inhibitor of both tryptophan and tyrosine hydroxylase. Thus, our results suggest that the observed hyperphenylalaninemia in patients who excrete 7-BH4 in their urine may arise directly from the inhibition of phenylalanine hydroxylase by low levels of this pterin. On the other hand, it is less likely that low levels of 7-BH4 would affect the activity of tyrosine or tryptophan hydroxylase in vivo.

Effects of nutritional status on contents of tryptophan, serotonin and 5-hydroxyindoleacetic acid in rat brain

Effects of nutritional status on the levels of tryptophan (Trp), serotonin (5HT), and its metabolite, 5-hydroxyindoleacetic acid (5HIAA) in six brain regions of rats were investigated. 1) A low-protein high-carbohydrate diet decreased Trp, 5HT and 5HIAA levels in the cortex and hippocampus, and those of 5HT and 5HIAA in the hypothalamus. This diet did not affect the contents of Trp and 5HT in the midbrain, but decreased 5HIAA. No significant changes of Trp, 5HT and 5HIAA were observed in the pons and medulla and striatum. 2) A low-carbohydrate high-protein diet increased the levels of Trp, 5HT and 5HIAA in the striatum, and 5HT and 5HIAA in the cortex, but showed no effect on the contents of Trp, 5HT and 5HIAA in the hippocampus, midbrain, pons and medulla and hypothalamus. 3) An energy-restriction low-carbohydrate diet increased Trp, 5HT and 5HIAA contents in the striatum, and 5HT and 5HIAA in the cortex and pons and medulla. In the hypothalamus, only 5HIAA was increased by this diet. The diet did not influence Trp, 5HT and 5HIAA contents of the midbrain and hippocampus. These results suggest that i) lowered fat and carbohydrate intakes enhance 5HT synthesis and metabolism in the cortex and that lowered carbohydrate intake enhances them in the striatum, ii) energy restriction enhances the 5HT metabolism in the cortex, pons and medulla and hypothalamus, and 5HT synthesis in the cortex and pons and medulla, iii) lowered protein intake inhibits 5HT metabolism in the cortex, midbrain, hippocampus and hypothalamus, and 5HT synthesis in the cortex, hippocampus and hypothalamus.

Localization of the specific binding sites of [3H]tryptophan in rat brain

The regional and cellular localizations of the previously reported aromatic amino acid binding sites (AABS) in rat brain were investigated using [(3)H]tryptophan as the binding radioligand. An almost 4-fold difference in AABS was observed among the brain regions in the descending order of cerebral cortex ? hippocampus > striatum ? thalamus/hypothalamus ? midbrain ? cerebellum ? brainstem. When rats were injected intracerebroventricularly (i.c.v.) with 5,7-dihydroxytryptamine (0.2 ?mol) after prior administration of desipramine (i.p., 25 mg/kg), [(3)H]tryptophan binding was significantly reduced by 32% in the brainstem but increased by 27% in the midbrain. After injection of 6-hydroxydopamine (i.c.v., 0.2 ?mol), however, binding was significantly reduced in the brainstem by 32%, striatum by 27% and cerebral cortex by 18% but increased by 24% in the midbrain. It is concluded that the AABS are associated with some but not all cells of the three monoaminergic systems in the rat brain. The increases in binding in the midbrain may reflect a localization of the AABS in this region in cells post-synaptic to the degenerating cells.