Precursor Of Serotonin Synthesis Research Articles

Differences in responses of premotor interneurons to serotonin and 5-hydroxytryptophan, a precursor for serotonin synthesis, in naive and trained snails

This study investigated the responses of premotor interneurons LPa3 and RPa3 of the snails to applications of serotonin and 5-hydroxytryptophan (5-HTP), a precursor for serotonin synthesis, as ingredients of the broth that bathes the central nervous system of naive snails and snails after the formation of long-term sensitization. Measurements of the electrical characteristics have shown that a membrane potential of interneurons LPa3 and RPa3 in naive snails was significantly depolarized (3.7 mV) in the presence of serotonin, while the threshold potential was increased (unsupported hypothesis). The similar pattern was observed in the presence of 5-hydroxytryptophan: true depolarization to 3.1 mV occurred in interneurons until reaching the unreliable rise in the threshold potential inducing an action potential. It was found that application of serotonin causes a significant decrease in the membrane potential of interneurons LPa3 and RPa3 of trained snails (depolarization to 4.6 mV) and the unreliable increase in the threshold potential of premotor interneurons (0.9 mV). In contrast, application of 5-hydroxytryptophan causes an unreliable increase (2.5 mV) of the membrane potential and also an unreliable increase (0.8 mV) in the threshold potential.

Evaluation of the anti-depressant potential of metformin in conditioned defeat model in golden Syrian hamsters

Background: Depression is a prevalent mood condition that has an impact on daily functioning. Globally, depression affects 264 million people. The current pharmacotherapy of depression has a lot of shortcomings. Therefore, there is a need to explore newer therapy that alleviate the symptoms of depression. Metformin was found to possess antioxidant potential and hypothesized to decrease the levels of branched-chain amino-acids essential for tryptophan uptake (precursor for serotonin synthesis). The study was designed to validate the efficacy of metformin as an anti-depressant in conditioned defeat model in male golden Syrian hamsters using open field test (OFT), forced swim test (FST) and Serum serotonin levels. Methods: After obtaining IAEC approval, the study was carried out using eight golden Syrian hamsters that were randomly assigned to four groups. The disease control group received 1mL normal saline, positive control was given fluoxetine 12 mg/kg, two groups of metformin 240 mg/kg given pre-insult and post-insult. The variables assessed on every third day included OFT and FST. Following the behavioral tests, serotonin-ELISA was done. To analyse the outcomes, appropriate statistical tests were applied. Results: On standardization, the model was established to a 16-day model. Further, results highlighted a significant difference in OFT, FST and serotonin levels with the metformin group and fluoxetine compared to disease control (p<0.001). However, no significant difference was observed between the fluoxetine and metformin groups (p>0.05), signifying the comparable results. Conclusions: Metformin (240 mg/kg) alleviated the depressive symptoms by modulating both behavioral and serotonin levels.

Physical exercise and 5-hydroxytryptophan, a precursor for serotonin synthesis, reduce penicillin-induced epileptiform activity

AimPrevious studies have shown that 5- hydroxytryptophan (5-HTP) and exercise play an important role in the synthesis of serotonin independently. The effects of the treadmill exercise and 5‐ hydroxytryptophan (5-HTP) on seizure mechanisms created by epileptiform activity with penicillin model were investigated in rats. MethodA total of 28 male albino Wistar rats were randomly divided into four groups: exercise (Ex), Control (Cnt), 5‐hydroxytryptophan (5htp) and 5-hydroxytryptophan + exercise (5htpEx) groups. Treadmill exercise and gavage (25 mg/kg/day) were administered five days a week for ten weeks. Electrocorticogram data were recorded for 3 h at the end of the protocol using the Power-Lab data acquisition system. Spike frequency, amplitude, and latency time were analyzed offline. The significant differences among the groups were evaluated by one-way analysis of variance (ANOVA). ResultsSpike frequency was observed at the highest level from the 20th minute in the Cnt group, and this continued until the end of the recording. The 5-HTP alone group did not affect epileptiform activity. At the 80th minute of penicillin injection, the epileptiform activity in the 5htpEx group decreased significantly compared with the Cnt, and this significance continued until the 110th minute. There was no statistical difference in the amplitude values of the groups. The 5htpEx group was significantly higher than both the Cnt and Ex group in the seizure latency times. ConclusionsIt was determined that exercise reduced the spike number and delayed seizure significantly by potentiating the effect of 5-HTP. Given that 5-HTP used in combination with exercise can perform useful actions such as reducing seizure sensitivity and consequently improving the quality of life for individuals with epilepsy, it may be a potential candidate for the treatment of epilepsy in nonpharmacological methods.

5-Hydroxytryptophan, a precursor for serotonin synthesis, reduces seizure-induced respiratory arrest.

The DBA/1 mouse is a relevant animal model of sudden unexpected death in epilepsy (SUDEP), as it exhibits seizure-induced respiratory arrest (S-IRA) evoked by acoustic stimulation, followed by cardiac arrhythmia and death. Defects in serotonergic neurotransmission may contribute to S-IRA. The tryptophan hydroxylase-2 (TPH2) enzyme converts L-tryptophan to 5-hydroxytryptophan (5-HTP), a precursor for central nervous system (CNS) serotonin (5-HT) synthesis; and DBA/1 mice have a polymorphism that decreases TPH2 activity. We, therefore, hypothesized that supplementation with 5-HTP may bypass TPH2 and suppress S-IRA in DBA/1 mice. TPH2 expression was examined by Western blot in the brainstem of DBA/1 and C57BL/6J mice both with and without acoustic stimulation. Changes in breathing and cardiac electrical activity in DBA/1 and C57BL/6J mice that incurred sudden death during generalized seizures evoked by pentylenetetrazole (PTZ) were studied by plethysmography and electrocardiography. The effect of 5-HTP administration on seizure-induced mortality evoked by acoustic stimulation or by PTZ was investigated in DBA/1 mice. Repetitive acoustic stimulation resulted in reduced TPH2 protein in the brainstem of DBA/1 mice as compared with C57BL/6J mice. S-IRA evoked by acoustic stimulation in DBA/1 mice was significantly reduced by 5-HTP. Following S-IRA, cardiac electrical activity could be detected for minutes before terminal asystole and death in both DBA/1 and C57BL/6J mice after PTZ treatment. The incidence of S-IRA by PTZ administration was greater in DBA/1 than in C57BL/6J mice, and administration of 5-HTP also significantly reduced S-IRA by PTZ in DBA/1 mice. Our data suggest that S-IRA is the primary event leading to death incurred in most DBA/1 and some C57BL/6J mice during PTZ-evoked seizures. Suppression of S-IRA by 5-HTP suggests that 5-HT transmission contributes to the pathophysiology of S-IRA, and that 5-HTP, an over-the-counter supplement available for human consumption, may be clinically useful in preventing SUDEP.

Tryptophan Transport in Human Fibroblast Cells—A Functional Characterization

There are indications that serotonergic neurotransmission is disturbed in several psychiatric disorders. One explanation may be disturbed transport of tryptophan (precursor for serotonin synthesis) across cell membranes. Human fibroblast cells offer an advantageous model to study the transport of amino acids across cell membranes, since they are easy to propagate and the environmental factors can be controlled. The aim of this study was to functionally characterize tryptophan transport and to identify the main transporters of tryptophan in fibroblast cell lines from healthy controls.Tryptophan kinetic parameters (Vmax and Km) at low and high concentrations were measured in fibroblasts using the cluster tray method. Uptake of 3H (5)-L-tryptophan at different concentrations in the presence and absence of excess concentrations of inhibitors or combinations of inhibitors of amino acid transporters were also measured. Tryptophan transport at high concentration (0.5 mM) had low affinity and high Vmax and the LAT1 isoform of system-L was responsible for approximately 40% of the total uptake of tryptophan. In comparison, tryptophan transport at low concentration (50 nM) had higher affinity, lower Vmax and approximately 80% of tryptophan uptake was transported by system-L with LAT1 as the major isoform. The uptake of tryptophan at the low concentration was mainly sodium (Na+) dependent, while uptake at high substrate concentration was mainly Na+ independent. A series of different transporter inhibitors had varying inhibitory effects on tryptophan uptake.This study indicates that tryptophan is transported by multiple transporters that are active at different substrate concentrations in human fibroblast cells. The tryptophan transport trough system-L was mainly facilitated by the LAT1 isoform, at both low and high substrate concentrations of tryptophan.

Interactions between serotoninergic and aminoacidergic pathways in the control of PRL secretion in prepubertal male rats.

Prolactin secretion is controlled by the hypothalamus through different neurotransmitters which interact with multiple receptor subtypes. The discovery of different families of receptors for serotonin (5-HT1-5-HT7) and excitatory aminoacids (NMDA, KA, AMPA and metabotropic receptors) ilustrates the complexity of this regulation. Moreover, in the rat the role of different neurotransmitters changes during pubertal development. Present experiments were carried out to analyse the interactions between AMPA and serotoninergic receptors in the control of prolactin secretion in prepubertal male rats. For this purpose, 16 and 23-day old male rats were treated with 5-hydroxytryptophan (5-HTP, precursor of serotonin synthesis) plus fluoxetine (blocker of serotonin reuptake), 8-OH-DPAT (agonist of 5-HT1A receptors), DOI and alpha-Me-5-HT (agonists of 5-HT2 receptors), 1-phenylbiguanide (agonist of 5-HT3 receptors) alone or in combination with AMPA (agonist of AMPA receptors). The results obtained indicate that: (a) activation of 5-HT1A receptors stimulated PRL secretion on day 16 and inhibited it on day 23; activation of 5-HT2 receptors stimulated PRL secretion on days 16 and 23, whereas activation of 5-HT3 receptors inhibited PRL release only on day 23; (b) activation of AMPA receptors inhibited PRL secretion on day 23, but not on day 16 and (c) a cross-talk is apparent between 5-HT2 and AMPA receptors in the regulation of PRL secretion, the stimulatory effect of DOI being blocked by AMPA.

Effects of L-thyrosyl - L-arginine (kyotorphin) on the behavior of rats and goldfish

The effects of kyotorphin (KTP), a dipeptide (L-Tyr-L-Arg), on the level of sensory attention to stimuli of different modalities in rats and the exploratory behavior in goldfish were investigated. In both cases KTP was found to suppress the exploratory activity. When 5-HTP, a precursor of serotonin synthesis, is activated the inhibitory effects of KTP increased. It is assumed that the regulatory effect of KTP on the exploratory behavior of animals is mediated by the monoaminergic (neurotransmitting) brain systems, as distinct from its analgetic effects, which are mediated by the opioid brain systems.

Role of the serotoninergic system in the acceleration of sexual maturation in wild Norway rats selected for reduced aggressiveness toward humans

The role of the serotoninergic system in acceleration of the sexual development of domesticated rats ( Rattus norvegicus) was assessed. The onset of age-related changes in hypothalamic serotonin during prepubertal period occurred earlier in domesticated than in aggressive male rats. Blockade of the serotoninergic system after p-chlorophenylalanine (PCPA) administration on days 40 and 44 delayed the development of the reproductive system in both aggressive and domesticated males. In 60-day-old rats treated with PCPA, levels of testosterone in plasma and the number of mature spermatozoa in epididymis were decreased compared to controls. At the same time, the administration of PCPA on days 30 and 34 did not modify basal testosterone secretion and other parameters in 60-day-old aggressive rats and produced a decrease similar to PCPA injections on days 40 and 44, although less pronounced, in the weights of testes in domesticated animals. Administration of 5-hydroxytryptophan (5-HTP), a precursor of serotonin synthesis, on days 30, 32, 34, 36 and 38 increased plasma testosterone levels and weights of the sex organs in 60-day-old domesticated males, but did not significantly affect the development of reproductive system in aggressive animals. These data indicate that serotonin stimulates sexual development of males during prepubertal period and this activating effect of serotonin occurs earlier in domesticated than in aggressive males. They also suggest that the acceleration in sexual maturation of domesticated rats could result from changes in the ontogenetic dynamic of hypothalamic serotonin induced by a selection for low aggressiveness towards man.

Human brain serotonin synthesis capacity measured in vivo with alpha-[C-11]methyl-L-tryptophan.

Local cerebral serotonin synthesis capacity was measured with alpha-[C-11]methyl-L-tryptophan ([C-11]AMT) in normal adult human brain (n = 10; five males, five females; age range, 18-38 years, mean 28.3 years) by using positron emission tomography (PET). [C-11]AMT is an analog of tryptophan, the precursor for serotonin synthesis, and is converted to alpha-[C-11]methyl-serotonin ([C-11]AM-5HT), which is trapped in serotonergic neurons because [C-11]AM-5HT is not degraded by monoamine oxidase. Kinetic analysis of [C-11] activity in brain after injection of [C-11]AMT confirmed the presence of a compartment with unidirectional uptake that represented approximately 40% of the activity in the brain at 50 min after tracer administration. The undirectional rate constant K, which represents the uptake of [C-11]AMT from the plasma to brain tissue followed by the synthesis and physiologic trapping of [C-11]AM-5HT, was calculated using the Patlak graphic approach on a pixel-by-pixel basis, thus creating parametric images. The rank order of K values for different brain regions corresponded well to the regional concentrations of serotonin in human brain (P < .0001). High serotonin synthesis capacity values were measured in putamen, caudate, thalamus, and hippocampus. Among cortical regions, the highest values were measured in the rectal gyrus of the inferior frontal lobe, followed by transverse temporal gyrus; anterior and posterior cingulate gyrus; middle, superior, and inferior temporal gyri; parietal cortex; occipital cortex, in descending order. Values in women were 10-20% higher (P < .05, MANOVA) throughout the brain than those measured in men. Differences in the serotonin synthesis capacity between men and women measured in this study may reflect gender differences of importance to both normal and pathologic behavior. This study demonstrates the suitability of [C-11]AMT as a tracer for PET scanning of serotonin synthesis capacity in human brain and provides normal adult values for future comparison with patient groups.

Pyridoxine effect on synthesis rate of serotonin in the monkey brain measured with positron emission tomography.

The influence of the co-factor pyridoxine, vitamin B6, on the activity of aromatic amino acid decarboxylase enzyme was studied by positron emission tomography, PET in the brain of the Rhesus monkey using the precursor for serotonin synthesis 5-hydroxy-L-tryptophan (5-HTP) radiolabelled with 11C in the beta-position. The rate constant for the formation of serotonin in the corpus striatum was calculated using a two tissue compartment model with reference area in the brain. In baseline investigations, the mean rate constants (+/-S.D:) for selective utilization of [11C]5-HTP to form [11C]serotonin in the corpus striatum was 0.0080 +/- 0.0011 min(-1). Pretreatment with intravenous pyridoxine hydrochloride 10 mg/kg bodyweight before doing a second PET study resulted in an enhanced rate constant by a mean of 20%. The rate increase was statistically significant. The increase varied considerably in different monkeys from no effect to more than 60%. The effect of pyridoxine on aromatic amino acid decarboxylase activity supported a regulatory role of pyridoxine on the synthesis of neurotransmitter in vivo, and may be of importance in diseases with deficiencies in neurotransmitter function.

Role of the serotonergic system in the control of gonadotropin secretion in prepubertal male rats.

To analyze the role of brain serotonin in the control of gonadotropin secretion in prepubertal males, serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) concentrations were studied at different ages after administration of 5-hydroxytryptophan (5-HTP), a precursor of serotonin synthesis, alone or in combination with fluoxetine, a specific inhibitor of serotonin uptake; 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), a serotonin 1A agonist; or D,L-p-chlorophenylalanine methyl ester hydrochloride (PCPA), an inhibitor of serotonin synthesis. Also, serum gonadotropin concentrations were measured in castrated males implanted with silastic capsules containing testosterone or 5 alpha-androstane-3 alpha, 17 beta-diol (alpha-diol). We found that: (i) the blockade of the serotoninergic system after PCPA administration did not modify basal gonadotropin secretion but did strongly reduce the response to orchidectomy; (ii) a stimulatory effect of 5-HTP on FSH secretion was only observed in intact males at the age of 8 days; (iii) the serotonin 1A agonist 8-OH-DPAT did not change serum gonadotropin concentrations; (iv) 5-HTP increased serum FSH and LH concentrations in 16-day-old castrated males; (v) FSH and LH serum concentrations decreased after 5-HTP in castrated animals implanted with testosterone, whereas castrated animals implanted with alpha-diol showed, like the intact males, a lack of response to 5-HTP.(ABSTRACT TRUNCATED AT 250 WORDS)

An analysis of fear and aggression during early development of behaviour in silver foxes ( Vulpes vulpes)

The relationship between the development of behaviour in a novel situation and plasma cortisol level was analysed in 30-, 45- and 60-day-old domestic (D) foxes and foxes selected for enhanced aggressiveness (EA) towards man (Experiment 1). In the EA foxes, the increase in defensive responses (fear and aggression) was associated with a significant rise in plasma cortisol level at 45 and 60 days of age. The D foxes showed no defensive responses up to Day 60 and there were no associated changes in plasma cortisol level. In Experiment 2, chloditane, an inhibitor of adrenal cortex function, injected to the EA pups from Days 38–44 and 45–52 of life, attenuated fear and increased locomotor activity in the novel situation; there was an associated decline in plasma cortisol level. Treatment with chloditane, however, did not affect the number of foxes showing aggression in the novel situation and at the appearance of man both at the age of 60 days and in adulthood. Experiment 3 demonstrated a significant increase in locomotor activity in the novel situation and a decrease in the number of EA foxes exhibiting aggression after treatment with 1-tryptophan (a precursor of serotonin synthesis) after the age of 60 days. The EA foxes treated at an early age with 1-tryptophan showed a significant attenuation of aggression when adults. Taken together, the results demonstrate that plasma cortisol level and the fear response are related and also that enhanced aggressiveness affects greatly the development of behaviour in the EA foxes and contributes, together with the fear reaction, to limitation of the socialisation period.

Neuronal correlates of the serotonin-dependent behavior of the pteropod molluscClione limacina

A pharmacological model of hunting behavior, previously developed by the authors on the basis of the utilization of 5-hydroxytryptophan (5-HTP), a precursor of serotonin synthesis (5-HT), was employed in order to determine its cytochemical and neurophysiological correlates. In the presence of baseline behavior, 5-HT-containing neurons were not detected by the glyoxylate method, but they were readily identified in the buccal, cerebral, and pedal (the greatest number) ganglia of the CNS and in the peripheral organs during hunting behavior induced by an injection of 5-HTP (100-200 micrograms/g). The disappearance of pauses in the initial unsustained fictive swimming, an increase in the frequency of the rhythm of swimming and in the generation of spikes in each cycle, as well as a change in the phasing of the locomotor cycle, the neurophysiological correlates of "fast" locomotion in the presence of 5-HTP-induced hunting behavior, were recorded in isolated pedal ganglia in the presence of 5-HTP (100-200 micrograms/ml). The activity of 5-HT-neurons of the pedal ganglia, which was initially synchronized with the episodes of swimming, increases against the background of 5-HTP and becomes continuous. The effects of exogenous and endogenous 5-HT on the generator of locomotion are identical, and are induced by the direct action of the mediator on the membrane of the locomotor neurons. The hypothesis of the participation of 5-HT in the monoaminergic control of behavior is discussed.

Role of serotoninergic neurones in the control of gonadotrophin and prolactin secretion in the rat.

The role of brain serotonin (5-hydroxytryptamine, 5-HT) in the control of LH, FSH and prolactin secretion was studied in two groups of experimental animals: intact adult male rats and ovariectomized adult female rats. 5-Hydroxytryptophan (5-HTP), a precursor of serotonin synthesis, and fluoxetine, a specific inhibitor of 5-HT uptake, were given either alone or together. 5-Hydroxytryptophan (50 mg/kg) was administered intraperitoneally and fluoxetine (20 μg/rat) was given into one of the lateral ventricles of the brain. Neither 5-HTP nor fluoxetine given alone affected LH secretion but combined treatment with the two drugs elicited a significant increase in serum LH levels in both intact male and ovariectomized female rats. Fluoxetine and 5-HTP, alone or together, did not modify FSH secretion in either kind of animal. In intact males and in ovariectomized females, 5-HTP induced a significant increase in prolactin release; fluoxetine alone was ineffective. In male animals treated with fluoxetine plus 5-HTP, serum prolactin levels increased but such an increase was lower than that found in the animals treated only with 5-HTP. In ovariectomized rats, the combined treatment induced an increase in serum prolactin levels similar to that found in animals treated with 5-HTP alone. These data suggested that brain serotonin exerts a stimulating effect on LH secretion in both intact male and ovariectomized rats, but that it does not play any role in the control of FSH release in either kind of animal and that central serotoninergic pathways participate in the stimulating control of prolactin release from the anterior pituitary gland. However, some of the data also suggested the possibility of the existence in the brain of serotoninergic systems inhibiting prolactin secretion.