Levels Of 5-hydroxytryptophan Research Articles

Urinary Metabolite Markers of Precocious Puberty

The incidence of precocious puberty (PP, the appearance of signs of pubertal development at an abnormally early age), is rapidly rising, concurrent with changes of diet, lifestyles, and social environment. The current diagnostic methods are based on a hormone (gonadotropin-releasing hormone) stimulation test, which is costly, time-consuming, and uncomfortable for patients. The lack of molecular biomarkers to support simple laboratory tests, such as a blood or urine test, has been a long standing bottleneck in the clinical diagnosis and evaluation of PP. Here we report a metabolomic study using an ultra performance liquid chromatography-quadrupole time of flight mass spectrometry and gas chromatography-time of flight mass spectrometry. Urine metabolites from 163 individuals were profiled, and the metabolic alterations were analyzed after treatment of central precocious puberty (CPP) with triptorelin depot. A panel of biomarkers selected from >70 differentially expressed urinary metabolites by receiver operating characteristic and logistic regression analysis provided excellent predictive power with high sensitivity and specificity for PP. The altered metabolic profile of the PP patients was characterized by three major perturbed metabolic pathways: catecholamine, serotonin metabolism, and tricarboxylic acid cycle, presumably resulting from activation of the sympathetic nervous system and the hypothalamic-pituitary-gonadal axis. Treatment with triptorelin depot was able to normalize these three altered pathways. Additionally, significant changes in the urine levels of 4-hydroxyphenylacetic acid, 5-hydroxyindoleacetic acid, indoleacetic acid, 5-hydroxytryptophan, and 5-hydroxykynurenamine in the CPP group suggest that the development of CPP condition may involve an alteration in symbiotic gut microbial composition.

Measurement of 5-Hydroxytryptamine Turnover Rate in Rat Cerebral Arteries

Two methods for assessing 5-hydroxytryptamine (5-HT) turnover rate have been tested in the cerebral vessels of the rat. The pretreatment of the animals with benserazide 45 min before death brought about an increase in the levels of 5-hydroxytryptophan (5-HTP). When the MAO inhibitor pargyline was injected and the animals killed at different times, there was an exponential decrease in the concentration of 5-hydroxyindole acetic acid (5-HIAA) with time, whereas the increase in 5-hydroxytryptamine was linear only during the first 30 min, thereafter reaching a plateau. This pattern was similar to that obtained in the caudate nucleus after MAO blockade. In the hippocampus, pargyline induced a lineal accumulation of 5-HT throughout the experiment as well as an exponential decay of the 5-HIAA concentration. These results indicate that the turnover rate of 5-HT can be appraised in the rat cerebral arteries either after the rate of disappearance of 5-HIAA or from the accumulation rate of 5-HTP.

Effect and mechanism of senkyunolide I as an anti-migraine compound from Ligusticum chuanxiong

To evaluate the analgesic and anti-migraine activities of senkyunolide I from Ligusticum chuanxiong. Mice were orally administered various doses of senkyunolide I, and their pain levels were assessed in a hot-plate test and by application of acetic acid. The levels of 5-hydroxytryptamine (5-HT), 5-hydroxytryptophan (5-HTP), 5-hydroxyindoleacetic acid (5-HIAA), norepinephrine (NE) and dopamine (DA) in plasma and brain were assessed, and the monoamine turnover rates (5-HT/5-HTP, 5-HIAA/5-HT and NE/DA) were also calculated. Mice given senkyunolide I at 16 and 32 mg/kg had significantly elevated pain thresholds in the hot-plate test, and a dose of 32 mg/kg also reduced the number of abdominal writhing responses caused by acetic acid. Significant improvements were observed in the neurotransmitter levels of the drug-treated rats compared with the saline-administered controls. Compared to the rats with nitroglycerin-induced migraines, the levels of nitric oxide in the plasma and whole brain of rats given senkyunolide I were lower. The present study suggests that senkyunolide I may be an active component of L. chuanxiong, traditionally used to treat migraine. The mechanism of pain relief in migraine model rats may be through adjusting the levels of monoamine neurotransmitters and their turnover rates, as well as decreasing nitric oxide levels in the blood and brain. Therefore, senkyunolide I may be developed as a potential treatment for migraine pain.

Effects of marijuana and diazepam on lipid peroxidation, Na+, K+ATPase, and levels of glutathione and 5-HTP in rat brain

Our aim was to evaluate the effects of marijuana (Mar) and diazepam (Dz) on lipid peroxidation (TBARS), Na + , K + ATP ase activity, levels of glutathione (GSH) and 5-hydroxytryptophan (5-HTP). Male Wistar rats were given a single dose per group: extract of Mar (100 microL/kg), Dz (5 mg/kg), Mar plus Dz, and NaCl for control. Sixty mins after treatment, animals were sacrificed, and their brains extracted and homogenised to measure GSH, TBARS and 5-HTP levels. Na + , K + ATP ase and total ATP ase activities. GSH and TBARS did not show differences respect to controls. Na + , K + ATP ase activity was similar as well. However, groups treated with Mar, total ATPase activity decreased significantly (p < 0.05). Levels of 5-HTP decreased significantly (p = 0.0001) in rats treated either with Mar and or Dz. Mar and Dz induced biochemical effects on the serotonergic metabolism, which can alter the development and function in rat brain, because it has also been involved in scavenging free radicals present there.

Serum Levels of Tryptophan, 5-Hydroxytryptophan and Serotonin in Patients Affected with Different Forms of Amenorrhea

Tryptophan (Trp) is present in the serum, partly bound to albumine and in the free form. The unbound portion of circulating tryptophan has the property of crossing the hematoencephalic barrier and being converted within the brain into serotonin (5-HT) through the enzymatic processes of hydroxylation and decarboxylation. The serotoninergic system plays an important role in neuroendocrine control of reproductive hormone secretion, and in particular, it may influence GnRH pulsatility, a function essential for reproductive processes. In this study, we analysed serum levels of tryptophan, serotonin and 5-hydroxytryptophan (5-HTP) in women with three different forms of amenorrhea: 16 patients were diagnosed with anorexia nervosa, 60 patients with functional hypothalamic amenorrhea, and 14 patients with hyperprolactinemia. Data were compared with those of a group of 25 healthy women. Serum Trp levels were significantly (P ≤ 0.05) lower in the anorexic (11.64 ± 0.53 μg/ml, mean ± S.E.) than in the control (12.98 ± 0.37 μg/ml) groups. In addition, in the anorexic group a statistical dispersion of Trp values was shown indicating a bimodal data distribution suggesting the existence of two different subgroups of patients. Regarding 5-HTP, an increase of its serum level was observed in all the groups with amenorrhea with the highest value in hyperprolactinemic patients. On the contrary, no statistical differences in serum 5-HT levels among the four analyzed groups were observed.This study shows that women affected by various forms of amenorrhea present an altered metabolism of tryptophan via serotonin and, in particular, markedly high differences are observed between the two subgroups of anorexic patients.

Indole levels in human lumbar and ventricular cerebrospinal fluid and the effect of L-tryptophan administration.

Levels of tryptophan (TRP), 5-hydroxytryptophan (5-HTP), 5-hydroxytryptamine (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) in human lumbar and ventricular cerebrospinal fluid (CSF) were measured by reversed phase liquid chromatography (HPLC) with electrochemical detection. The levels of TRP ranged from 1593 to 4865 nmol/l in ventricular (VF) and from 1257 to 2557 nmol/l in lumbar CSF. The level of 5-HTP varied from 1.1 to 68.9 nmol/l in VF and from 5.3 to 10.8 nmol/l in lumbar CSF; no previous reports of 5-HTP levels in CSF exist. The serotonin level was 1.9-27.3 nmol/l in VF and 5.7-12.0 nmol/l in lumbar CSF. The levels of 5-HIAA were considerably higher in VF than in lumbar fluid with respective means of 498 +/- 52.4 nmol/l and 112 +/- 15.6 nmol/l (P less than 0.001). An oral dose of 2 g L-tryptophan significantly increased all indole levels except that of 5-HT, both in patients with progressive myoclonus epilepsy and in controls.

Pyridoxine, regardless of serotonin levels, increases production of 5-hydroxytryptophan in rat brain

Background The aim of this study was to evaluate effects of pyridoxine and butylated hydroxytoluene (BHT) on lipid peroxidation and on levels of 5-hydroxytryptophan and serotonin. Methods Thirty rats (30 days of age) were used in the survey, measuring levels of lipid peroxidation (TBARS), hemoglobin, 5-hydroxytryptophan (5-HTP), and serotonin (5-HT) after intraperitoneal (i.p.) injections of 4 and 10 mg/kg/day of pyridoxine HCl during 20 days and a single dose of 2 μM/kg (440 μg) of BHT. Results Levels of TBARS and 5-HTP increased considerably ( p <0.05) in all vitamin- and/or BHT-treated groups, and 5-HT increased partially ( p <0.05) only in B 6 with or without BHT-treated groups compared with control group. Conclusions Results suggest that pyridoxine plays a role in tryptophan metabolism, increasing production of 5-HTP.

Serum tryptophan and 5-hydroxytryptophan at birth and during post-partum days.

Serum levels of tryptophan (Trp) and 5-hydroxytryptophan (5-HTP) were measured in the mother at birth, by normal delivery or cesarean section, on days 1 and 3 post-partum, and in the cord blood of the fetus-newborns. Decreases in Trp and 5-HTP levels during delivery were noted. Conversely, Trp levels in the cord blood of fetus-newborns were significantly higher than those of their mothers, and 5-HTP levels also showed a tendency to increase , whereas on day 1 post-partum Trp values were lower than those in fetal cord, but higher than those during spontaneous delivery. On day 3 post-partum, serum Trp levels were higher than those during spontaneous delivery, but similar to those on day 1. 5-HTP values tended to decrease with respect to those of day 1. Serum Trp and 5-HTP concentrations decreased significantly on day 1 post-partum after cesarean section, but increased on day 3, demonstrating that maternal Trp was being consumed during delivery. A major release of prolactin in serum on days 1 and 3 post-partum there was in women with normal delivery, revealing a correlation between Trp and prolactin in the first few days after birth, in relation to different conditions of delivery.

Acute effects of L-tryptophan on tryptophan hydroxylation rate in brain regions (hypothalamus and medulla) of rainbow trout (Oncorhynchus mykiss).

Levels of 5-hydroxytryptophan (5-HTP) in brain regions (hypopthalamus and medulla) of rainbow trout were analysed by HPLC-EC 0, 10, 30, and 40 min after intraperitoneal administration of different doses of L-tryptophan (Trp) (0, 12.5, and 25 mg. kg(-1) body weight) in fish treated with 3-hydroxybenzylhydrazine (NSD1015; 75 mg. kg(-1)). The results show that, in control fish, 5-HTP levels in hypothalamus (58.03 +/- 6.36 pg. mg(-1) brain tissue) were significantly higher than those observed in medulla (28.04 +/- 4.32 pg. mg(-1) brain tissue). Basal tryptophan hydroxylation rates (after 0 mg. kg(-1) Trp administration) were 0.42 +/- 0.07 pg 5-HTP. mg(-1). min(-1), and 0.63 +/- 0.24 pg 5HTP. mg(-1). min(-1), for hypothalamus and medulla respectively. On the other hand, the results demonstrate that L-tryptophan administration induced significant increases in the rate of tryptophan hydroxylation, both in hypothalamus and medulla. These findings indicate that, in a way similar to that observed in mammals, brain tryptophan hydroxylase is unsaturated by its substrate (tryptophan) under normal physiological conditions. J. Exp. Zool. 286:131-135, 2000.

Effects of Fusarium mycotoxins on levels of serotonin, melatonin, and 5-hydroxytryptophan in pineal cell cultures.

Analysis of melatonin (MEL) in pineal cell cultures by enzyme linked immunosorbent assay showed its concentration was increased by fusaric acid (FA), a mycotoxin produced by Fusarium species and associated with toxic duck and ostrich feeds. Subsequent cell culture studies demonstrated the precursors of MEL, 5-hydroxytryptophan (5HTP) and serotonin (5HT), were also affected by FA as well as other Fusarium mycotoxins. Herein we describe a technique for the analysis of 5HTP and 5HT in pineal cell cultures using HPLC with electrochemical detection (EC), and report on the effects of FA alone and in combination with fumonisin B1 (FB1) and deoxynivalenol (DON) on the levels of these MEL precursors.

Changes in nocturnal pineal indoleamine metabolism in rats treated with parathion are prevented by β-adrenergic antagonist administration

Parathion, an organophosphorous insecticide, was previously shown to enhance the nighttime rise in pineal N-acetyltransferase (NAT) activity and serum melatonin levels. The purpose of the present study was to test whether parathion acts on the pineal gland by means of a β-adrenergic receptor mechanism. Whereas parathion (total dose 6.5 mg/kg body wt over 6 days) by itself significantly augmented nocturnal pineal NAT activity and serum melatonin levels in otherwise untreated rats, the insecticide was ineffective in reference to this enzyme when it was given in conjunction with the β-adrenergic receptor antagonist propranolol (20 mg/kg body wt, l h before lights off). The augmentation of NAT activity by parathion also caused significant reductions in pineal serotonin (5-HT); again, this response was blocked by propranolol treatment. Neither pineal hydroxyindole- O-methyltransferase (HIOMT) activity nor pineal levels of 5-hydroxytryptophan or hydroxyindole acetic acid (5-HIAA) were significantly changed as a result of either parathion or propranolol treatment. The results are consistent with the idea that parathion influences pineal S-HT metabolism either at the level of the β-adrenergic receptor or via the sympathetic innervation to the pineal gland.

Studies on biogenic amines and their metabolites in nervous tissue and hemolymph of adult male cabbage looper moths—I. Quantitation of photoperiod changes

HPLC with electrochemical detection was used to quantify levels of 5-hydroxytryptamine, octopamine, dopamine, tryptophan, tyramine, tyrosine, 5-hydroxytryptophan, 5-hydroxyindolacetic acid, 3,4-dihydroxyphenylacetic acid, N-acetyl-5-hydroxytryptamine, N-acetyloctopamine, and N-acetyldopamine in optic lobes, corpora allata/corpora cardiaca compex, thoracic ganglia, supraesophageal-subesophageal ganglion complex (or brain) and hemolymph in individual male cabbage looper moths, Trichoplusia ni, at five times in the photoperiod. Levels of amines remained unchanged throughout the photophase, but decreased in some tissues during scotophase. In the hemolymph, levels of octopamine increased during the initial part of the dark period, whereas levels of dopamine remained unchanged, and 5-hydroxytryptamine was not detected. Levels of tryptophan, tyrosine, 5-hydroxytrytophan and tyramine did not change significantly over the photoperiod. Levels of N-acetyloctopamine, N-acetylserotonin and N-acetyldopamine in each tissue increased corresponding to the observed decreases in octopamine, 5-hydroxytryptamine, or dopamine. No evidence was found for catabolic activity involving monoamine oxidase. Within individuals, there was no correlation between decreased levels of octopamine and 5-hydroxytryptamine in the brain, or with levels of either of these amines in the thoracic ganglia. However, decreases in octopamine in the thoracic ganglia paralleded decreases in dopamine. The results are discussed with respect to the proposed modulatory action of amines on male moth locomotor activity and response to sex pheromone.

Effects of oxiracetam on organic brain syndrome following activation of the high-affinity choline uptake system: A neurochemical analysis of the cerebrospinal fluid of four patients

Nootropic drugs stimulate acetylcholine (ACh) synthesis by activating the high-affinity choline uptake system. Cerebrospinal fluid (CSF) samples were obtained from four patients with organic brain syndrome in order to gauge acetylcholinesterase activity and measure levels of ACh, 5-hydroxyindoleacetic acid, vanillylmandelic acid, and 5-hydroxytryptophan before and at the end of nootropic treatment. Patients were given oxiracetam for 30 days at dosages varying from 1600 mg/day to 2000 mg/day. Clinical evaluations conducted at the beginning and end of treatment were made by measuring the Sandoz Clinical Assessment—Geriatric (SCAG) score and reaction times to simple auditory and visual stimuli. Following oxiracetam treatment, average ACh levels in CSF decreased significantly, and there was a concomitant improvement in the clinical evaluation (SCAG).

Brain indoles in human hepatic encephalopathy

The neurotransmitter serotonin has a profound effect on the control of sleep; thus excess serotonin activity in the brain could be responsible for impaired consciousness in hepatic encephalopathy. Furthermore, an increased brain level of 5-hydroxyindoleacetic acid has been a consistent finding in various animal models of the condition. In this study, using high-performance liquid chromatography with fluorometric detection, we examined levels of brain serotonin (5-hydroxytryptamine) and its precursors and metabolites in 16 patients dying with hepatic encephalopathy complicating acute and chronic liver disease and 9 control subjects matched for age, sex, postmortem delay in brain retrieval and length of frozen tissue storage. In patients with chronic liver disease, serotonin level was significantly increased in thalamus (p < 0.02); levels of its metabolite 5-hydroxyindoleacetic acid were increased in frontal cortex (p < 0.05), globus pallidus (p < 0.05) and putamen (p < 0.01). Levels of the precursor amino acid tryptophan were increased in thalamus (p < 0.01) and globus pallidus (p < 0.01); in both patient groups levels of 5-hydroxytryptophan and the tryptamine metabolite indoleacetic acid were increased in all brain areas studied. 5-Hydroxytryptamine levels were also increased in thalamus, frontal cortex and globus pallidus in the three patients with fulminant liver failure. Our findings are consistent with disordered neurotransmission, especially in the thalamus, an area of particular importance in the regulation of consciousness, alertness and attention in human beings. (HEPATOLOGY 1993;17:1033–1040.)

Brain indoles in human hepatic encephalopathy

The neurotransmitter serotonin has a profound effect on the control of sleep; thus excess serotonin activity in the brain could be responsible for impaired consciousness in hepatic encephalopathy. Furthermore, an increased brain level of 5-hydroxy-indoleacetic acid has been a consistent finding in various animal models of the condition. In this study, using high-performance liquid chromatography with fluorometric detection, we examined levels of brain serotonin (5-hydroxytryptamine) and its precursors and metabolites in 16 patients dying with hepatic encephalopathy complicating acute and chronic liver disease and 9 control subjects matched for age, sex, postmortem delay in brain retrieval and length of frozen tissue storage. In patients with chronic liver disease, serotonin level was significantly increased in thalamus (p < 0.02); levels of its metabolite 5-hydroxyindoleacetic acid were increased in frontal cortex (p < 0.05), globus pallidus (p < 0.05) and putamen (p < 0.01). Levels of the precursor amino acid tryptophan were increased in thalamus (p < 0.01) and globus pallidus (p < 0.01); in both patient groups levels of 5-hydroxytryptophan and the tryptamine metabolite indoleacetic acid were increased in all brain areas studied. 5-Hydroxytryptamine levels were also increased in thalamus, frontal cortex and globus pallidus in the three patients with fulminant liver failure. Our findings are consistent with disordered neurotransmission, especially in the thalamus, an area of particular importance in the regulation of consciousness, alertness and attention in human beings.

Studies on CSF tryptophan metabolism in infantile spasms

Cerebrospinal fluid from 8 patients with infantile spasms (mean age: 6.1 months) was collected before treatment. The concentration of cerebrospinal fluid tryptophan metabolites was analyzed using high-performance liquid chromatography and compared to metabolite concentrations in cerebrospinal fluid from 20 age-matched controls (mean age: 5.8 months). The levels of cerebrospinal fluid serotonin, 5-hydroxyindoleacetic acid, and kynurenine were significantly lower in infantile spasm patients compared to controls ( P < .05). In contrast, the levels of cerebrospinal fluid 3-hydroxykynurenine were significantly higher in infantile spasm patients than in controls ( P < .05). There were no significant differences in the levels of cerebrospinal fluid tryptophan and 5-hydroxytryptophan. Although the study population was small, these findings suggest that the presence of seizures in infantile spasms is associated with a decrease in serotonergic metabolites which, in turn, may indicate a decrease in serotonergic activity, altered clearance of these metabolites, or altered turnover in the direction of 3-hydroxykynurenine. The perturbance caused by increased 3-hydroxykynurenine and decreased kynurenine in the homeostatic balance between these 2 tryptophan metabolites could further contribute to the pathogenesis of infantile spasms.

Circadian rhythm of tryptophan hydroxylase activity in chicken retina

1. Retinal tryptophan hydroxylase activity in chickens (1-4 weeks old and embryos) was estimated by determination of levels of 5-hydroxytryptophan (5HTP) in retinas at defined intervals after inhibition of aromatic L-amino acid decarboxylase with m-hydroxybenzylhydrazine (NSD1015). 2. The relationship of tryptophan hydroxylase activity to photoperiod was explored. In chickens maintained on a 12-hr light: 12-hr dark cycle, a diurnal cycle in tryptophan hydroxylase activity was observed. Activity during middark phase was 4.4 times that seen in midlight phase. Cyclic changes in tryptophan hydroxylase activity persisted in constant darkness with a period of approximately 1 day, indicating regulation of the enzyme by a circadian oscillator. The phase of the tryptophan hydroxylase rhythm was found to be determined by the phase of the light/dark cycle. The relationship of the tryptophan hydroxylase rhythm to the light/dark cycle mirrors previously described rhythms of melatonin synthesis and serotonin N-acetyltransferase (NAT) activity in the retina. 3. Light exposure for 1 hr during dark phase suppressed NAT activity by 82%, while tryptophan hydroxylase activity was suppressed by only 30%. 4. Based on the differential responses of retinal NAT activity and tryptophan hydroxylase activity to acute light exposure during dark phase, it was predicted that exposure to light during dark phase would divert serotonin in the retina from melatonin biosynthesis to oxidation by MAO. In support of this, levels of 5-hydroxyindole acetic acid (5HIAA) in retina were found to be elevated approximately two-fold in chickens exposed to 30 min of light during dark phase. In pargyline-treated chickens, 2 hr of light exposure during dark phase was found to increase retinal serotonin levels by 64% over pargyline-treated controls. 5. Cyclic changes in tryptophan hydroxylase activity and NAT activity persisted for 2-3 days in constant light. Tryptophan hydroxylase activity at mid-night gradually decreased on successive days in constant light; on the first day of constant light, tryptophan hydroxylase activity at mid-night was 70% of activity seen during middark phase of the normal light/dark cycle and decreased further on subsequent days. In contrast, on each of 3 days of constant light, NAT activity at mid-night was approximately 15% of normal middark phase activity. 6. Cycloheximide completely inhibited the nocturnal increase in tryptophan hydroxylase activity when given immediately before light offset. The nocturnal increase in NAT activity was inhibited in a similar fashion. 7. Like the development of the NAT rhythm, cyclic changes of tryptophan hydroxylase activity in the retinas of chickens began on or immediately before the day of hatching. hatching.(ABSTRACT TRUNCATED AT 400 WORDS)

Parathion (O,O-dimethyl-O-p-nitrophenyl phosphorothioate) induces pineal melatonin synthesis at night

The effects of parathion on male rat pineal N-acetyltransferase (NAT) activity, hydroxyindole-O-methyltransferase (HIOMT) activity and pineal and serum melatonin levels at the end of light period (2000 h) and at night (2300 h and 0100 h) were studied. Additionally, pineal levels of 5-hydroxytryptophan (5-HTP), serotonin (5-HT), and 5-hydroxyindole acetic acid (5-HIAA) were estimated. Parathion was administered intragastrically at total doses (over 6 days) of either 6.5 or 13 mg/kg. Control rats received vehicle (corn oil) only. During the study, the rats were exposed to light:dark cycles of 14:10 with light off at 2100 h. Pineal NAT activity was increased at 0100 h following parathion administration at both doses, but HIOMT activity was unaffected. Pineal and serum melatonin levels were increased at night (2300 h and 0100 h) after the 13 mg/kg dose of parathion while the lower dose increased pineal melatonin only at 0100 h. Also, both doses decreased 5-HTP at 2000 h while the lower dose increased it at 2300; 5-HT was significantly decreased at 2300 h and 5-HIAA levels were lower but only significantly so for the 13 mg/kg dose at 2000 h. The results indicate that parathion has significant effects on pineal melatonin synthesis by mechanisms which remain unknown.

Determination of melatonin and monoamines in rat pineal using reversed-phase ion-interaction chromatography with fluorescence detection

A method is reported for the ion-interaction, reversed-phase separation of 24 compounds (chiefly monoamines) arising from the metabolism of tyrosine and tryptophan. These compounds were separated as two groups. The first group comprised 3,4-dihydroxyphenylethylene glycol, tyrosine, 3-methoxy-4-hydroxyphenyl glycol, 5-hydroxytryptophan, norepinephrine, 3,4-dihydroxyphenylacetic acid, epinephrine, 5-hydroxyindole-3-acetic acid, homovanillic acid, 5-hydroxytryptophol, dopamine, tryptophan. N-acetylserotonin, N-acetyltryptophan, 5-methoxytryptophan and serotonin. The mobile phase consisted of a 6.8:93.2 (v/v) mixture of acetonitrile and an aqueous solution containing 0.16 M ammonium phosphate, 0.06 M citric acid, 0.15 mM disodium EDTA, 10 mM dibutylamine and 6 mM sodium 1-octanesulphonate at pH 4.50. The second group of compounds comprised 6-hydroxymelatonin, 5-methoxyindole-3-acetic acid, indole-3-acetic acid, 5-methoxytryptamine, tryptamine, 5-methoxytryptophol, melatonin and tryptophol. The mobile phase consisted of a 16:84 (v/v) mixture of acetonitrile and an aqueous solution containing 0.05 M ammonium phosphate, 0.05 M citric acid, 0.15 mM disodium EDTA, 25 mM dibutylamine and 5 mM sodium 1-octanesulphonate at pH 5.30. Detection was by fluorescence measurement (lambda ex = 280 nm, lambda em = 340 nm). The proposed method exhibited linear calibration over the biochemically significant concentration range, with detection limits in the 10-200 pg range. Excellent precision for peak areas and retention times was observed, even over a period of 24 h. The applicability of amperometric detection (at 0.72V) is also demonstrated. The method is applied to the determination of monoamines in individual rat pineals. Low nanogram levels of tyrosine, norepinephrine, 5-hydroxyindole-3-acetic acid, tryptophan, serotonin and 6-hydroxymelatonin, and picogram levels of 5-hydroxytryptophan, 5-hydroxytryptophol, 5-methoxyindole-3-acetic acid, indole 3-acetic acid, 5-methoxytryptophol and melatonin were indicated in most of the samples.

Carbaryl-induced changes in indoleamine synthesis in the pineal gland and its effects on nighttime serum melatonin concentrations

The effects of different doses of chronically administered carbaryl on rat pineal N-acetyltransferase (NAT) activity, hydroxyindole- O-methyltransferase (HIOMT) activity and pineal and serum melatonin levels during darkness (2300 h and 0100 h) when pineal melatonin synthesis is high were studied. Additionally, pineal levels of 5-hydroxytryptophan (5-HTP), serotonin (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) were estimated. Carbaryl was administered at total doses (over 6 days) of either 50, 125 or 250 mg/kg by gastric gavage. Control rats received vehicle (corn oil) only. During the study, the rats were exposed to light/dark cycles of 14:10 with lights off at 2100 h. Pineal NAT and HIOMT activities and pineal melatonin were increased at 0100 h following carbaryl administration at all three doses. Conversely, serum melatonin was increased at 2300 h after the 250 mg/kg dose of carbaryl while all three doses of the pesticide reduced serum melatonin levels at 0100 h. Pineal 5-HTP, 5-HT and 5-HIAA levels were usually increased at 2300 h but unaffected at 0100 h. The results indicate that carbaryl has significant effects on pineal melatonin synthesis and secretion.