In-vivo measurements of tryptophan and tyrosine hydroxylase activities in mouse brain

Summary A single intraperitoneal injection of NSD 1015 (3-hydroxybenzyl-hydrazine HCl, 100 to 200 mg/kg), an inhibitor of the aromatic amino acid decarboxylase, caused an initially linear accumulation of 5-hydroxy-tryptophan (5-HTP) and 3, 4-dihydroxyphenylalanine (DOPA) in mouse brain. The initial rate of accumulation of 5-HTP after NSD 1015 and that of 5-hydroxytryptamine (5-HT) after treatment with the monoamine oxidase inhibitor pargyline were almost identical (about 1.7 nmoles/g·h^–1 corresponding to a turnover time of 1.6 h) and may represent the true rate of 5-HT synthesis. About 60 min after pargyline, when the 5-HT level was doubled, the rate of 5-HT synthesis was partially inhibited, probably due to feedback inhibition.The observations on DOPA accumulation after NSD 1015 were in essential agreement with earlier rat brain data and suggest that the rate constants of catecholamine synthesis are similar in the brains of the two species.     

Microdialysis with radiometric monitoring of -[β-11C]DOPA to assess dopaminergic metabolism: effect of inhibitors of -amino acid decarboxylase,monoamine oxidase,and catechol-O-methyltransferase on rat striatal dialysate

The catecholamine, dopamine (DA), is synthesized from 3,4-dihydroxy--phenylalanine (-DOPA) by aromatic -amino acid decarboxylase (AADC). Dopamine metabolism is regulated by monoamine oxidase (MAO) and catechol-O-methyltransferase (COMT). To measure dopaminergic metabolism, we used microdialysis with radiometric detection to monitor -[β-¹¹C]DOPA metabolites in the extracellular space of the rat striatum. We also evaluated the effects of AADC, MAO, and COMT inhibitors on metabolite profiles. The major early species measured after administration of -[β-¹¹C]DOPA were [¹¹C]3,4-dihydroxyphenylacetic acid ([¹¹C]DOPAC) and [¹¹C]homovanillic acid ([¹¹C]HVA) in a 1:1 ratio, which shifted toward [¹¹C]HVA with time. An AADC inhibitor increased the uptake of -[β-¹¹C]DOPA and -3-O-methyl-[¹¹C]DOPA and delayed the accumulation of [¹¹C]DOPAC and [¹¹C]HVA. The MAO and COMT inhibitors increased the production of [¹¹C]3-methoxytyramine and [¹¹C]DOPAC, respectively. These results reflect the -DOPA metabolic pathway, suggesting that this method may be useful for assessing dopaminergic metabolism.     

Kinetic compartment modeling of [11C]-5-hydroxy-L-tryptophan for positron emission tomography assessment of serotonin synthesis in human brain.

The substrate for the second enzymatic step in serotonin synthesis, 5-hydroxy-L-tryptophan, labeled in the beta-position ([11C]-HTP), was used for positron emission tomography (PET) measurements in six healthy human participants, examined on two occasions. One- and two-tissue kinetic compartment modeling of time-radioactivity curves was performed, using arterial, metabolite-corrected [11C]-HTP values as input function. The availability of unchanged tracer in arterial blood plasma was > or = 80% up to 60 minutes after injection, while [11C]-hydroxyindole acetic acid and [11C]-serotonin accounted for the remaining radioactivity, amounting to < or = 16% and < or = 4%, respectively. Compartment modeling was performed for brain stem, putamen, caudate nucleus, anterior cingulate, white matter, and superior occipital, occipitotemporal, and temporal cortices. The average biologic half-life for plasma-to-tissue equilibrium was 7 to 12 minutes, and the volume of distribution was 0.2 to 0.5 microL.mL(-1). In all regions except white matter, the kinetic compartment model that included irreversible [11C]-HTP trapping showed significantly improved model fits with respect to a one-tissue compartment model. The [11C]-HTP trapping rate constant depended on the estimated tissue availability of the serotonin precursor tryptophan, known to reflect serotonin synthesis in healthy individuals, and correlated with serotonin tissue concentration and synthesis rates reported previously in literature. These findings suggest the use of [11C]-HTP PET measurements to investigate serotonin synthesis.     

5-Hydroxytryptamine is synthesized in neurons terminating in the neural and intermediate lobes of the rat pituitary gland

Immunocytochemistry has revealed that nerve fibers within the neural and intermediate lobes of the rat pituitary gland contain 5-hydroxytryptamine (5-HT). Recent anatomical evidence suggests that the content of this amine in the intermediate but not the neural lobe of the pituitary gland may represent 5-HT that has been taken up from the blood rather than synthesized intraneuronally. The purpose of this study was to determine if 5-HT is synthesized in neurons of the neurointermediate lobe of the pituitary gland. 5-HT synthesis was estimated by measuring the accumulation of the 5-HT precursor, 5-hydroxytryptophan (5-HTP), in the neurointermediate lobe of male Long-Evans rats following the administration of NSD 1015, an inhibitor of aromatic L-amino acid decarboxylase. Thirty min following the injection of NSD 1015 (100 mg/kg, i.p.), 5-HTP accumulated in the neurointermediate lobe and the rate of this accumulation was increased by the administration of the 5-HTP precursor, tryptophan, and by electrical stimulation of the pituitary stalk. In addition, repeated injections of the 5-HT uptake inhibitor, fluoxetine (10 mg/kg, i.p., every 12 h for a total of 7 injections), induced a marked depletion of platelet 5-HT but did not alter the concentration of 5-HT in either the neural or intermediate lobes of the pituitary gland. Taken together these results indicate that much of the 5-HT in the neurointermediate lobe of the pituitary gland does not represent 5-HT taken up from the blood, but rather the amine is synthesized in neurons projecting to this region.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of increased endogenous serotonin on the in vivo binding of [11C]DASB to serotonin transporters in conscious monkey brain

Using a combination of positron emission tomography (PET) and the microdialysis technique, the effects of increased endogenous serotonin (5-hydroxytryptamine; 5-HT) on the binding of [(11)C]DASB to 5-HT transporters (5-HTT) were investigated in the conscious monkey brain. Five rhesus monkeys (Macaca mulatta) were scanned with [(11)C]DASB under the control condition and the increased endogenous 5-HT condition, in which 5-hydroxy-L-tryptophan (5-HTP) was administered (20 mg/kg, i.v.) before the PET scan. Compared with the control scan, the 5-HTP administration significantly decreased the binding potential (BP) (BP = B(max)/K(d)) of [(11)C]DASB in several brain regions. The mean % decrease of BP was biggest in the caudate and putamen. Two monkeys were scanned with [(11)C]5-HTP to assess the amino acid decarboxylase (AADC) activity in the brain, resulting in the high activity in the caudate and putamen. Microdialysis measurements showed that although 5-HTP administration (20 mg/kg, i.v.) increased the extracellular 5-HT levels in both the prefrontal cortex and caudate, the increase of the 5-HT level in the caudate was 27 times higher than that in the prefrontal cortex. These results suggest that the caudate and putamen, both of which show high AADC activity, convert 5-HTP to 5-HT at a high rate, and the increased 5-HT competes with [(11)C]DASB for the 5-HTT.     

Validation studies on the 5-hydroxy-L-[beta-11C]-tryptophan/PET method for probing the decarboxylase step in serotonin synthesis

The two-tissue compartment model, including irreversible trapping in the second compartment (2TCM) is used to describe the kinetics of 5-Hydroxy-L-[beta-(11)C]-tryptophan ([(11)C]HTP), a radioligand used in positron emission tomography (PET) for probing the second enzymatic step in the biosynthesis of serotonin. In this study, we examined the capacity of the model to track pharmacological changes in this biological process. We also investigated the potential loss of [(11)C]HTP-derived radioactivity during a PET study, since loss should be negligible not to alter quantification. Six rhesus monkeys were investigated using bolus [(11)C]HTP/PET methodology before and after pharmacological intervention. The second enzymatic step in serotonin synthesis was inhibited using the aromatic L-amino acid decarboxylase inhibitor NSD1015 (10 mg/kg). The extent of [(11)C]-derived radioactivity loss from the brain was studied by inhibition of the enzyme responsible for formation of the tissue metabolite, monoamine oxidase A, using clorgyline (2 mg/kg). After NSD1015, the uptake of [(11)C]HTP-derived radioactivity was increased in all the investigated brain regions, while the parameter used to reflect decarboxylase activity, the net accumulation rate constant (K(acc)), was decreased by 37% in the striatum, compared with baseline. Pretreatment with clorgyline did not change the brain uptake of [(11)C]HTP-derived radioactivity or K(acc). This study demonstrates that the 2TCM for [(11)C]HTP/PET is able to detect changes occurring during alteration of the biological process (i.e., the conversion of HTP to serotonin). Elimination of the radiotracer metabolite [(11)C]HIAA from the brain may be considered negligible if the PET study is limited to 60 min.     

The yohimbine-induced anticonflict effect in the rat,part II. Neurochemical findings

Summary In a companion paper the ₂-adrenoceptor antagonist yohimbine was found to produce a dose-dependent anticonflict effect in a modified Vogel's conflict test. The behavioral data further indicated that noradrenergic and serotonergic neurons as well as the benzodiazepine (BDZ) receptor may be involved in the anticonflict effect of yohimbine. In the present study the effects on rat brain monoamine neurochemistry and GABA _a /BDZ receptor function (³⁶Cl^–-uptake in corticohippocampal synaptoneurosomes) of a maximally anticonflict producing dose of yohimbine (4.0 mg/kg, i.p.) were studied. The levels of rat brain catecholamines and indoleamines were measured ex vivo using high performance liquid chromatography with electrochemical detection (HPLC-ED). Yohimbine decreased noradrenaline levels both in the hippocampus and the hemispheres but instead increased DOPAC levels in these brain regions as well as in the limbic forebrain. Yohimbine also markedly enhanced DOPA accumulation in the hippocampus and the hemispheres after inhibition of 1-aromatic amino acid decarboxylase by means of NSD 1015, whereas in the limbic system only a modest increase was obtained. The yohimbine-induced effects on the catecholamine synthesis rate were largely abolished in animals severely depleted of NA by means of 6-hydroxy-dopamine (6-OH-DA) pretreatment. Yohimbine decreased both the 5-HIAA/5-HT quotient (an indicator of 5-HT turnover) and 5-HTP accumulation after NSD 1015 in the hemispheres, whereas in the hippocampus and the limbic system only 5-HTP accumulation was decreased. The yohimbine-induced effect on the indoleamine synthesis rate was not influenced by 6-OH-DA pretreatment, whereas this effect and that on the catecholamine synthesis rate were both abolished by reserpine pretreatment. Neither in vivo nor in vitro administration of yohimbine significantly altered baseline or GABA-induced accumulation of³⁶Cl^– in corticohippocampal synaptoneurosomes. In conclusion, the present study provides neurochemical support for the suggestion that yohimbine may exert its anticonflict effect in a modified Vogel's conflict test by increasing and decreasing NA and 5-HT neurotransmission, respectively, whereas no evidence was obtained for a direct interaction of yohimbine with GABA _a /BDZ receptor function.     

Evaluation of [18F]-JNJ-64326067-AAA tau PET tracer in humans

The [¹⁸F]-JNJ-64326067-AAA ([¹⁸F]-JNJ-067) tau tracer was evaluated in healthy older controls (HCs), mild cognitive impairment (MCI), Alzheimer’s disease (AD), and progressive supranuclear palsy (PSP) participants. Seventeen subjects (4 HCs, 5 MCIs, 5 ADs, and 3 PSPs) received a [¹¹C]-PIB amyloid PET scan, and a tau [¹⁸F]-JNJ-067 PET scan 0-90 minutes post-injection. Only MCIs and ADs were amyloid positive. The simplified reference tissue model, Logan graphical analysis distribution volume ratio, and SUVR were evaluated for quantification. The [¹⁸F]-JNJ-067 tau signal relative to the reference region continued to increase to 90 min, indicating the tracer had not reached steady state. There was no significant difference in any bilateral ROIs for MCIs or PSPs relative to HCs; AD participants showed elevated tracer relative to controls in most cortical ROIs (P < 0.05). Only AD participants showed elevated retention in the entorhinal cortex. There was off-target signal in the putamen, pallidum, thalamus, midbrain, superior cerebellar gray, and white matter. [¹⁸F]-JNJ-067 significantly correlated (p < 0.05) with Mini-Mental State Exam in entorhinal cortex and temporal meta regions. There is clear binding of [¹⁸F]-JNJ-067 in AD participants. Lack of binding in HCs, MCIs and PSPs suggests [¹⁸F]-JNJ-067 may not bind to low levels of AD-related tau or 4 R tau.     

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

Summary 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) radiola-belled with¹¹C in the -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.     

The binding of [3H]-5-hydroxytryptamine to homogenates of human brain

Summary The binding of [³H]-5-hydroxytryptamine ([³H]-5-HT) to homogenates of human brain has been studied. The specific binding is saturable, with a K_d (frontal cortex) of 12±2nM, and is inhibited by non-radioactive 5-HT (IC₅₀=26 nM) and D-Lysergic acid diethylamide (IC₅₀=20 nM). Specific, but not non-specific binding of [³H]-5-HT was inhibited by incubation of the homogenates at 50 °C. The binding of [³H]-5-HT across the human brain was not uniform, the highest binding being found in the substantia nigra and hippocampus, and the lowest in the thalamus and pons. The K_d of the binding sites towards 5-HT did, however, appear to be similar for the different brain regions.     

Uptake and utilization of [beta-11C]5-hydroxytryptophan in human brain studied by positron emission tomography.

The immediate precursor in the serotonin synthetic route, 5-hydroxytryptophan (5-HTP), labeled with 11C in the beta position, has become available for studies using positron emission tomography (PET) to examine serotonin formation in human brain. Normalized uptake and intracerebral utilization of tracer amounts of [beta-11C]5-HTP were studied twice in six healthy male volunteers, three of them before and after pharmacological pretreatments. The kinetic model defines regional utilization as the relative regional radioactivity accumulation rate. Repeat studies showed good reproducibility. Pretreatments with benserazide, p-chlorophenylalanine (PCPA), and unlabeled 5-HTP all significantly increased uptake of [beta-11C]5-HTP. The utilization rates in both striatal and frontal cortex were higher than those in the surrounding brain, indicating that PET studies using [beta-11C]5-HTP as a ligand quantitate selective processes in the utilization of 5-HTP. We tentatively interpret uptake and utilization as a measure of brain serotonin turnover, the selectivity of which was shown by pharmacological interventions in vivo.     

Effect of thiamine deficiency on brain serotonin turnover

Serotonin turnover has been investigated in regional brain areas of rats made thiamine deficient by pyrithiamine (PT). Following intracisternal injection of [¹⁴C]5-hydroxytryptamine ([¹⁴C]5-HT), a marked increase in the accumulation of [¹⁴C]5-hydroxyindoleacetic acid ([¹⁴C]5-HIAA) was found in the medulla-pons, hypothalamus and cerebral cortex. [¹⁴C]5-HT levels were normal in all of the brain areas except the cerebral cortex which had an increase of 58%. The ratio of [¹⁴C]5-HIAA/[¹⁴C]5-HT was significantly increased in every brain region of PT-treated rats except the cerebral cortex. Part of this increase in [¹⁴C]5-HIAA was shown to be due to impairment of active transport of this 5-HT metabolite out of the brain. However, increased 5-HT synthesis in the cerebellum, hypothalamus, striatum, hippocampus and cerebral cortex was demonstrated by measurement of 5-HT accumulation after inhibition of brain monoamine oxidase. PT-induced increase in endogenous 5-HIAA in the medulla-pons occurred simultaneously with the onset of neurological signs and both parameters were reversible by thiamine administration. These results suggest that acute thiamine deficiency, induced by PT, both increases brain 5-HT synthesis and impairs 5-HIAA efflux from the brain. There is a close correlation between neurological manifestations and changes in brain 5-HT metabolism in acute thiamine deficiency.     

Determination of the source of 5-hydroxytryptaminergic neuronal projections to the neural and intermediate lobes of the rat pituitary gland through the use of electrical stimulation and lesioning experiments

5-Hydroxytryptamine (5-HT)-containing axons and terminals have been visualized in the neural and intermediate lobes of the rat pituitary gland, but the origin of these fibers remains in question. This study was designed to determine if 5-HT cell bodies in the brainstem or in the dorsomedial nucleus of the hypothalamus project to either of these pituitary lobes. Since lesions and electrical stimulation of 5-HT cell bodies decrease and increase, respectively, the rate of 5-HT synthesis in regions innervated by these cells, these techniques were employed. The in vivo rate of 5-HT synthesis was determined by quantifying the rate of accumulation of the 5-HT precursor, 5-hydroxytryptophan (5-HTP), in the neural and intermediate lobes of the pituitary gland 30 min after the administration of a decarboxylase inhibitor (NSD 1015, 100 mg/kg, i.p.). The application of 30 min of stimulating current (monophasic cathodal pulses of 1 ms duration and 0.3 mA current delivered at a frequency of 10 Hz) to electrodes implanted in the dorsal and median raphe nuclei increased the rate of 5-HT synthesis in both the neural and intermediate lobes of the pituitary gland. 5,7-Dihydroxytryptamine lesions of these nuclei altered neither 5-HTP accumulation nor 5-HT concentrations in the neural and intermediate lobes, but similar lesions of the nuclei raphe pontis and raphe magnus decreased both the concentration of 5-HT and the accumulation of 5-HTP in these pituitary regions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mapping human brain fatty acid amide hydrolase activity with PET

Endocannabinoid tone has recently been implicated in a number of prevalent neuropsychiatric conditions. [¹¹C]CURB is the first available positron emission tomography (PET) radiotracer for imaging fatty acid amide hydrolase (FAAH), the enzyme which metabolizes the prominent endocannabinoid anandamide. Here, we sought to determine the most suitable kinetic modeling approach for quantifying [¹¹C]CURB that binds selectively to FAAH. Six healthy volunteers were scanned with arterial blood sampling for 90 minutes. Kinetic parameters were estimated regionally using a one-tissue compartment model (TCM), a 2-TCM with and without irreversible trapping, and an irreversible 3-TCM. The 2-TCM with irreversible trapping provided the best identifiability of PET outcome measures among the approaches studied (coefficient of variation (COV) of the net influx constant K_i and the composite parameter λk₃ (λ=K₁/k₂) <5%, and COV(k₃)<10%). Reducing scan time to 60 minutes did not compromise the identifiability of rate constants. Arterial spin labeling measures of regional cerebral blood flow were only slightly correlated with K_i, but not with k₃ or λk₃. Our data suggest that λk₃ is sensitive to changes in FAAH activity, therefore, optimal for PET quantification of FAAH activities with [¹¹C]CURB. Simulations showed that [¹¹C]CURB binding in healthy subjects is far from a flow-limited uptake.     

Measurement of 5-hydroxytryptamine synthesis and metabolism in selected discrete regions of the rat brain using high performance liquid chromatography and electrochemical detection: Pharmacological manipulations

High performance liquid chromatography coupled with electrochemical detection (LCEC) was employed to measure 5-hydroxytryptamine (5-HT), 5-hydroxyindole-3-acetic acid (5-HIAA) and 5-hydroxytryptophan (5-HTP) in the suprachiasmatic (SCN), medial preoptic (MPO) and arcuate (AN) nuclei as well as the median eminence (ME) and striatum (ST) of individual rat brains. Biochemical estimations of changes in 5-HT neuronal activity were made by measuring: (1) concentrations of 5-HT and 5-HIAA and (2) the rate of 5-HT synthesis (5-HTP accumulation following the administration of NSD 1015, an inhibitor of aromatic L-amino acid decarboxylase) after the administration of pharmacological agents known to influence these neurons. Pargyline increased the concentration of 5-HT and decreased the concentration of 5-HIAA while probenecid increased the concentration of 5-HIAA in all 5 brain regions. At both 2 and 24 hours after reserpine the concentration of 5-HT decreased, 5-HIAA increased or did not change, and the rate of 5-HT synthesis increased. In most of the brain regions blockers of 5-HT neuronal uptake (fluoxetine, chlorimipramine) did not influence 5-HT or 5-HIAA concentrations dramatically, but increased the rate of 5-HT synthesis. L-tryptophan generally increased the concentrations of 5-HT and 5-HIAA as well as the rate of accumulation of 5-HTP in all regions except the ME where 5-HIAA and 5-HTP concentrations both were unaffected. These results reveal that the method using LCEC is sensitive enough to detect pharmacologically-induced changes in 5-HT metabolism and synthesis in discrete regions of rat brain. The drugs examined in the present study generally caused similar changes in 5-HT dynamics in all 5 brain regions examined.     

Potentiating effects of L-type Ca2+ channel blockers on pentobarbital-induced hypnosis are influenced by serotonergic system

Summary. In order to elucidate the mechanism(s) behind the interactions between barbiturates and Ca²⁺ antagonists, the effects of three structurally diverse types of Ca²⁺ antagonists combined or not with 5-HT on pentobarbital-induced hypnosis in mice were investigated. The results showed that dihydropyridine derivative nifedipine (10.0 and 20.0 mg/kg, p.o.) and other types of Ca²⁺ antagonist, verapamil (5.0 and 10.0 mg/kg, p.o.) and diltiazem (2.5, 5.0 and 10.0 mg/kg, p.o.) increased both the sleeping time in hypnotic dosage of pentobarbital (45 mg/kg, i.p.) treated mice and the rate of sleep onset in the sub-hypnotic dosage of pentobarbital (28 mg/kg, i.p.) treated mice in a dose-dependent manner, respectively, and these effects were significantly augmented by 5-hydroxytryptophan (5-HTP), the immediate precursor of 5-hydroxytryptamine (5-HT). Pretreatment with p-chlorophenylalanine (PCPA, 300 mg/kg, s.c.), an inhibitor of tryptophan hydroxylase, significantly decreased pentobarbital-induced sleeping time and nifedipine (10.0 mg/kg, p.o.), verapamil (5.0 mg/kg, p.o.) and diltiazem (2.5 mg/kg, p.o.) abolished this effect. From these results, it should be presumed that the augmentative effect of L-type Ca²⁺ channel blockers on pentobarbital-induced sleep may be influenced by serotonergic system.     

Ex vivo inhibitory effect of the 5-HT uptake blocker citalopram on 5-HT synthesis

Summary Serotonin (5-hydroxytryptamine, 5-HT) synthesis was determined in vivo by measuring the accumulation of 5-hydroxytryptophan (5-HTP) in rat frontal cortex after inhibition of aromatic amino acid decarboxylase by administration of m-hydroxybenzylhydrazine (NSD 1015) (100 mg/kg, i.p.). The selective 5-HT reuptake inhibitor, citalopram, the 5-HT_1a agonists, (±)8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), ipsapirone, gepirone and the 5-HT_1a/b agonist, 7-trifluoromethyl-4(4-methyl-1-piperazinylpyrolo[1,2-a]-quinoxaline (CGS 12066B), the 5-HT_1a/b ligands and -adrenoceptor antagonists, (±)pindolol and (±)alprenolol, and the non-selective 5-HT ligands, m-chlorophenylpiperazine (mCPP) and metergoline, all inhibited the synthesis of 5-HT. The 5-HT_1a /5-HT₂ antagonist, spiperone, alone, had no effect on basal 5-HT synthesis, however it attenuated the effect of 8-OH-DPAT by 56% and CGS 12066B by 39% but only barely that of citalopram by 17%. The selective 5-HT_1a antagonist, WAY 100635, which did not modify by itself 5-HT synthesis, had no effect on citalopram-induced reduction of 5-HT synthesis. Neither the 5-HT₂ agonist, (±)1-(2,5-dimethoxy-4-indophenyl)-2-aminopropane (DOI) nor the 5-HT₂ antagonist, ritanserin, had any effect on the synthesis of 5-HT. In addition, ritanserin did not modify the inhibitory effect of citalopram. Methiothepin was the only compound to increase 5-HT synthesis. These results suggest that the effect of citalopram on the synthesis of 5-HT is not mediated by 5-HT_1a or 5-HT₂ receptors and that other receptors may be involved.     

Effect of gammahydroxybutyric acid on serotonin synthesis,concentration and metabolism in the developing rat brain

Summary 5-HT synthesis, levels and degradation were investigated in the whole brain and regional brain areas in 4,14, and 28 days old rats after administration of gammahydroxybutyric acid (GHBA). 5-HT synthesis was investigated by means of 5-HTP accumulation after decarboxylase inhibition by NSD 1015. 5-HTP accumulation increased in the 14 and 28 days old rats but decreased in the 4 days old animals 90 min after GHBA, 750 mg/kg. In the 28 days old rats a corresponding increase was also noted in the precursor amino acid tryptophan. Regional and whole brain 5-HT levels were not altered by GHBA treatment. Regional as well as whole brain levels of 5-HIAA increased in the 14 and 28 days old rats after GHBA administration.In conclusion, the present data indicate that GHBA increases the synthesis and degradation of 5-HT in adolescent rats. These effects of GHBA were not seen in the neonatal animals.     

Kinetic modeling without accounting for the vascular component impairs the quantification of [11C]PBR28 brain PET data

The positron emission tomography radioligand [¹¹C]PBR28 targets translocator protein (18 kDa) (TSPO) and is a potential marker of neuroinflammation. [¹¹C]PBR28 binding is commonly quantified using a two-tissue compartment model and an arterial input function. Previous studies with [¹¹C]-(R)-PK11195 demonstrated a slow irreversible binding component to the TSPO proteins localized in the endothelium of brain vessels, such as venous sinuses and arteries. However, the impact of this component on the quantification of [¹¹C]PBR28 data has never been investigated. In this work we propose a novel kinetic model for [¹¹C]PBR28. This model hypothesizes the existence of an additional irreversible component from the blood to the endothelium. The model was tested on a data set of 19 healthy subjects. A simulation was also performed to quantify the error generated by the standard two-tissue compartmental model when the presence of the irreversible component is not taken into account. Our results show that when the vascular component is included in the model the estimates that include the vascular component (2TCM-1K) are more than three-fold smaller, have a higher time stability and are better correlated to brain mRNA TSPO expression than those that do not include the model (2TCM).     

A possible mechanism of the nucleus accumbens and ventral pallidum 5-HT1B receptors underlying the antidepressant action of ketamine: a?PET study with macaques

Ketamine is a unique anesthetic reagent known to produce various psychotic symptoms. Ketamine has recently been reported to elicit a long-lasting antidepressant effect in patients with major depression. Although recent studies provide insight into the molecular mechanisms of the effects of ketamine, the antidepressant mechanism has not been fully elucidated. To understand the involvement of the brain serotonergic system in the actions of ketamine, we performed a positron emission tomography (PET) study on non-human primates. Four rhesus monkeys underwent PET studies with two serotonin (5-HT)-related PET radioligands, [¹¹C]AZ10419369 and [¹¹C]DASB, which are highly selective for the 5-HT1B receptor and serotonin transporter (SERT), respectively. Voxel-based analysis using standardized brain images revealed that ketamine administration significantly increased 5-HT1B receptor binding in the nucleus accumbens and ventral pallidum, whereas it significantly reduced SERT binding in these brain regions. Fenfluramine, a 5-HT releaser, significantly decreased 5-HT1B receptor binding, but no additional effect was observed when it was administered with ketamine. Furthermore, pretreatment with 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(f)quinoxaline (NBQX), a potent antagonist of the glutamate α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor, blocked the action of ketamine on the 5-HT1B receptor but not SERT binding. This indicates the involvement of AMPA receptor activation in ketamine-induced alterations of 5-HT1B receptor binding. Because NBQX is known to block the antidepressant effect of ketamine in rodents, alterations in the serotonergic neurotransmission, particularly upregulation of postsynaptic 5-HT1B receptors in the nucleus accumbens and ventral pallidum may be critically involved in the antidepressant action of ketamine.