Serotonin platelet-transporter measures in childhood attention-deficit/hyperactivity disorder (ADHD): clinical versus experimental measures of impulsivity.

Impulsivity in terms of aggression, suicide or poor cognitive control has been associated with low synaptic availability of serotonin (5-HT) in adults and children. However, characteristics of the 5-HT transporter have not been studied in children with attention-deficit/hyperactivity disorder (ADHD: combined type), where impulsivity is a core symptom. Here, we explored in 20 children with ADHD the relationship of the density (Bmax) and affinity (Kd) of the platelet 5-HT transporter measured with [3H]paroxetine to both clinical ratings of impulsivity (Conners' Parent Questionnaire), and an experimental measure of impulsivity (the ability to withhold a prepotent response in the "stop-signal" paradigm). Decreases of affinity (increased Kd) correlated with a low probability of response inhibition, but not with the clinical ratings of impulsivity. However, ratings of distractibility and impulsivity correlated with the experimental measure of response-inhibition. In contrast, increased transporter affinity (low Kd) correlated modestly with higher ratings of aggressive and externalising behaviour. Bmax was not associated with any behavioural score. We conclude that the synaptic availability of 5-HT is under the control of the 5-HT transporter binding site affinity and that low affinity may be related to cognitive impulsivity (distractibility). Increased affinity of the transporter may also be related to conduct disturbance.     

Serotonergic markers in platelets of patients with major depression: upregulation of 5-HT2 receptors.

The uptake of [3H]5-HT and the density (Bmax) as well as affinity (Kd) of 5-HT uptake sites labelled with [3H]paroxetine and of 5-HT2 receptors labelled by [3H]LSD were determined in platelets from 25 medication-free patients with major depression and 20 normal controls. The density (Bmax) of 5-HT2 receptors was found to be significantly increased (by 52%) in platelets from depressed patients, particularly females. No changes were found in the affinity (Kd) of 5-HT2 receptors and in 5-HT uptake or [3H]paroxetine binding parameters. Density of 5-HT2 receptors positively correlated with that of [3H]paroxetine sites in control but not in depressed subjects. No correlation was found between the HAMD scores and Bmax of [3H]LSD binding. The results suggest that upregulation of platelet 5-HT2 receptors is a useful biological marker in major depression, particularly in females.     

Laterality and 3H-imipramine binding: studies in the frontal cortex of normal controls and suicide victims.

Serotonin [5-hydroxytryptamine (5-HT)]-sensitive imipramine binding (IB) was determined in the left and right hemisphere of frontal cortex of suicide victims and nonpsychiatric controls who died due to myocardial infarction or accident. The Kd (an inverse measure of affinity of 3H-imipramine to its binding sites) was significantly higher in left hemisphere than right hemisphere in normal controls. There were no differences in Bmax and Kd or Bmax between left hemisphere and right hemisphere in normals and suicides, respectively. These results do not support the finding of hemispheric asymmetry of 5-HT uptake as measured by IB (Bmax) in postmortem tissue from controls and suicide victims.     

Platelet 5-HT2 serotonin receptor binding sites in autistic children and their first-degree relatives

We examined platelet serotonin2 [5-hydroxytryptamine2 (5-HT2)] receptor binding sites, whole blood serotonin (5-HT), and plasma norepinephrine (NE) in male autistic children and their first-degree relatives. Saturation studies utilizing 125I-spiroperidol labeled the 5-HT2 sites with an affinity of 224.6 +/- 84.4 pmol/L (Kd). No group differences, i.e., autistic (n = 12), siblings (n = 6), parents (n = 22), control (adult; n = 7: child; n = 10), were seen for either the Kd or the total number of sites (Bmax: 14.3 +/- 10.9 fmol/mg protein). No correlations were found in any group between binding parameters (Kd or Bmax) and whole blood 5-HT. For the parental group, inverse correlations were found between NE and Bmax (standing NE, rs = -0.67, n = 21, p = 0.001; supine NE, rs = -0.49, n = 22, p = 0.021). In the autistic group, no correlation was seen between plasma NE and Bmax. A correlation between the autistic boys' Bmax and their fathers' Bmax was observed (rs = 0.79, n = 11, p = 0.004). These findings suggest (1) circulating NE may be involved in heterologous regulation of 5-HT2 receptors in the platelet and (2) genetic (paternal-filial) factors may play a role in the expression of 5-HT2 binding sites in the platelet. These preliminary findings are discussed in relation to heterologous receptor regulation. The relationships between these findings and either the pathophysiology of autism or hyperserotonemia in autism are unknown.     

Serotonin function, personality-trait variations, and childhood abuse in women with bulimia-spectrum eating disorders

BACKGROUND: Across populations, findings associate impulsivity, behavioral disinhibition, or hostility with reduced central serotonin (5-hydroxytryptamine: 5-HT) activity and increased likelihood of childhood abuse. Inconsistently, findings associate compulsivity, behavioral inhibition, or anxiousness with elevated 5-HT neurotransmission. We explored relationships among measures of 5-HT system functioning, behavioral inhibition/disinhibition, and childhood abuse in women with bulimia-spectrum eating syndromes. METHOD: In 73 bulimic (body mass index [kg/m2] under 30, binge eating at least once weekly) and 50 normal-eater control women, we obtained indices of platelet paroxetine binding and 5-HT agonist (m-CPP)-stimulated neuroendocrine responses. Cluster analysis was used to classify the bulimic women according to 5-HT "profiles." Resulting groups were then compared on symptom and trait measures. RESULTS: Measures of paroxetine-binding density (Bmax) and affinity (Kd) contributed significantly (p < .001 and p < .02, respectively) to a classification of bulimic women into groups with "low density/high affinity" (N = 52) or "high density/low affinity" (N = 21) binding. The 5-HT based classification did not predict eating-symptom severity. However, the "high density" pattern was associated with increased perfectionism and compulsivity, reduced risk of childhood sexual abuse, and (to some extent) reduced probability of borderline personality disorder. DISCUSSION: In women with bulimic syndromes, serotonergic factors, personality-trait variations, and developmental typologies converge in principled fashion. Our findings corroborate (with neurobiological evidence) the concept of underregulated and overregulated subtypes within the bulimic population.     

Influence of age, sex and diurnal variability on imipramine receptor binding and serotonin uptake in platelets of normal subjects.

Imipramine (IMI) binding and serotonin (5-HT) uptake were determined in platelets of 98 healthy volunteers; and their association with age, sex and circadian rhythm were evaluated. A large interindividual variability was found for both IMI and 5-HT parameters. There was a negative correlation of IMI affinity constant (Kd) and binding (Bmax) with age, but no such correlation of 5-HT affinity constant (Km) or uptake (Vmax). Significant age-related diurnal variability was found for 5-HT Km in the whole group as well as for IMI Kd in males, but not in females. There was no significant correlation between 5-HT Vmax and IMI Bmax. Our results underscore a cautious approach to the interpretation of platelet serotonergic studies. In light of the multiple variables influencing the results, the usefulness of IMI or 5-HT as clinical markers should be re-evaluated.     

Dopamine may be 'hyper' with respect to noradrenaline metabolism, but 'hypo' with respect to serotonin metabolism in children with attention-deficit hyperactivity disorder

Noradrenaline: Hechtman (J Psychiat Neurosci 1994;19:193) argued for a role for frontal dopamine (DA) and noradrenaline (NA) in ADHD, where Oades (Prog Neurobiol 1987;29:365) has described lateralised functional impairments. Mechanisms (e.g. via alpha-2 sites) for stimulating low NA activity in ADHD children (J Am Acad Child Adolesc Psychiatry 1997;36:1688) in order to promote interactions with mesocortical DA have been discussed (J Psychopharmacology 1997;11:151; Psychiatr Res 1994;52:305). We described with indicators of overall transmitter metabolism (monoamines, metabolites in 24 h urine samples (Behav Brain Res 1997;88:95)) significantly lower utilisation ratios (MHPG/NA) in ADHD children with respect to healthy controls. Interestingly, a comparison of between catecholamine levels (DA/NA) showed a correlation with the conditioned blocking measure of selective attention recorded at the time of collection. This measure was negatively associated with blocking in controls. These results are consistent with reports of lower DOPEG and increased DOPAC in ADHD urine (J Child Adolesc Psychopharmacol 1996;6:63) and indicate that the relatively hyperactive DA versus NA systems may have functional consequences. Serotonin: the relevance for ADHD of an association of impulsivity with low serotonin (5-HT) metabolism (Behav Brain Sci 1986;9:319) has long been played down. Yet, some symptoms have been related to CSF measures of the metabolite 5-HIAA, and in particular the HVA/5-HIAA ratio has been reported to correlate with ratings of activity (Psychiatr Res 1994;52:305). We find that while urinary measures of 5-HIAA are somewhat higher, the ratio HVA/5-HIAA is markedly lower in ADHD children versus controls. In these ADHD children 5-HIAA levels were negatively related to d-prime measures in a continuous performance task (CPTax), and the HVA/5-HIAA was negatively associated with conditioned blocking. These results suggest a relatively low DA versus 5-HT activity may have functional consequences, albeit in a subgroup of ADHD. This is consistent with drug-induced prolactin changes reported by Verbaten et al. (Eur Child Adolesc Psychiatry 1999;8:30).     

Serotonin uptake and imipramine binding in the blood platelets of obsessive-compulsive disorder patients.

14C-Serotonin (5-HT) uptake and 3H-imipramine binding (IB) were studied in the blood platelets of 20 obsessive-compulsive disorder (OCD) patients, 53 normal controls (5-HT uptake) and 32 normal controls (IB binding). The maximum number of binding sites (Bmax) was significantly decreased in OCD patients compared to normal controls, but there was no difference in the affinity for 3H-imipramine (Kd). The affinity for 5-HT uptake (Km) was also decreased in the OCD patients but the maximum velocity of 5-HT uptake sites (Vmax) was not significantly different in OCD patients and normal volunteers. There were trends for the Slowness Subscale of the Maudsley Obsessional-Compulsive Inventory (MOCI) to be positively correlated with the Km of 5-HT uptake (p = 0.094), whereas the Global Scale, Checking Subscale, and Doubting Conscientiousness Subscale of MOCI were negatively correlated with the Kd of IB (p = 0.066, p = 0.08, and p = 0.062, respectively). The results provide further evidence for the dysfunction of the serotonergic system in OCD.     

Can serotonin transporter genotype predict serotonergic function,chronicity, and severity of drinking?

Serotonin transporter (5-HTT) activity is greater in carriers of the long (L) vs. short (S) alleles of the 5-HTT-linked polymorphic region (5'-HTTLPR) among healthy control subjects but not alcohol-dependent adults. In 198 alcoholics, we determined the relationship between current or lifetime drinking and platelet 5-HTT function and density among allelic variants of the 5'-HTTLPR. SS subjects were younger than L-carriers (LL and LS) (p<0.0085) and had fewer years of lifetime drinking. For L-carriers, the mean of Bmax for paroxetine binding, but not Vmax for serotonin (5-HT) uptake, was lower than that for SS subjects (p<0.05). More L-carriers than their SS counterparts had Vmax for 5-HT uptake below 200 nmol/10(7) platelets-min (p<0.05) and Bmax for paroxetine binding below 600 nmol/mg protein (p<0.06). Current drinking (drinks per day during the past 14 days) correlated positively with Km and Vmax of platelet 5-HT uptake (p<0.05) and negatively with Bmax, but not Kd, of paroxetine binding (p<0.05) for L-carriers alone. Years of lifetime drinking correlated negatively with Km and Vmax of platelet 5-HT uptake (p<0.05) and B(max), but not Kd, of paroxetine binding (p<0.05) for L-carriers alone. Among L-carriers alone, there were higher levels of platelet 5-HT uptake and lower levels of platelet paroxetine binding with increased drinking, and more lifetime drinking was associated with modestly lower levels of 5-HT uptake and paroxetine binding. Thus, 5-HTT expression varies with current and lifetime drinking in L-carriers alone.     

Platelet imipramine binding in autistic subjects

Previous reports of elevated platelet serotonin (5-HT) concentrations in autistic subjects suggest that platelet 5-HT uptake might be altered in autism. Parameters of 3H-imipramine (IMI) binding were measured in 11 drug-free autistic subjects and 10 normal volunteers. Similar means (+/- SD) for Bmax (autistics, 1350 +/- fmole/mg protein; normals, 1590 +/- 206 fmole/mg protein) and Kd (autistics, 0.98 +/- 0.10 nM; normals, 0.94 +/- 0.13 nM) were found in the two groups. The normal number (Bmax) and affinity (Kd) of the IMI binding site in autistic subjects suggest that the regulation of 5-HT uptake is not different in autism.     

Platelet vesicular monoamine transporter 2 density in the disruptive behavior disorders

In a former study, we reported decreased platelet vesicular monoamine transporter 2 (VMAT2) density (Bmax) in patients with ADHD. The current study aimed at measuring platelet VMAT2 in the disruptive behavior disorders (DBDs) to assess whether this finding is specific to ADHD or generalizable to the broader DBD concept. The study included 13 patients with DBDs aged 10-12 years and 16 healthy volunteers aged 8-17 years. All participants underwent a thorough clinical evaluation using Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime version for diagnosis, the Nisonger Child Behavior Rating Form, the Clinical Global Impressions Scale-Severity version, and the DSM-IV ADHD Scale (DAS). The study group's DAS scores did not differ from those of the control group. There was no significant difference between the patients with DBDs and the control group either in VMAT2 density (Bmax) or affinity (Kd) as measured by high-affinity [(3)H]TBZOH binding. We conclude that the formerly reported decreased platelet VMAT2 Bmax in patients with ADHD may be specific to ADHD and not present in DBDs. Larger-scale replication is needed.     

High affinity [3H]paroxetine binding to serotonin uptake sites in human brain tissue

[3H]Paroxetine binding to human brain tissue was characterized. Competition studies in the putamen and frontal cortex revealed single-site binding models for binding sensitive to 5-hydroxytryptamine (5-HT) (Ki 1-3 microM) and citalopram (Ki 0.6 nM), which displaced the same amount of binding. However, desipramine, norzimeldine and fluoxetine displaced additional binding (10-20%) and these competitors fitted two-site binding models with high affinity components in the nanomolar range and low affinity components in the micromolar range. The high affinity components approximated the 5-HT- and citalopram-sensitive binding fraction. Most of the [3H]paroxetine binding sites were protease-sensitive, but the low-affinity (microM) sites appeared to be protease-resistant. Based on these findings, only the [3H]paroxetine binding representing the fraction sensitive to 30 microM 5-HT (or e.g. 0.3 microM norzimeldine), was regarded as specific binding. This binding fraction was saturable with an apparent binding affinity (Kd) of 0.03-0.05 nM throughout the brain. The highest binding densities were obtained in the hypothalamus and substantia nigra (Bmax 500 fmol/mg protein). The basal ganglia reached intermediate densities (Bmax 200 fmol/mg protein), whereas cortical areas had low Bmax values (less than 100 fmol/mg protein). The lowest B max value was noted in cerebellar cortex (30 fmol/mg protein). The [3H]paroxetine binding was competitively inhibited by low concentrations of 5-HT, imipramine and norzimeldine, suggesting that the substrate recognition site for 5-HT uptake was labeled. Compounds active at dopaminergic, noradrenergic, histaminergic, 5-HT1, 5-HT2 and cholinergic muscarinic sites did not affect the binding at 100 microM concentrations. It is concluded that [3H]paroxetine is a marker for the 5-HT uptake site in the human brain, provided that an adequate pharmacological definition of specific binding is performed.     

Factorial validity,reliability of assessments and prevalence of ADHD behavioural symptoms in day and residential treatment centres for children with behavioural problems

This study uses the attention deficit/hyperactivity disorder (ADHD) symptom ratings of professional care workers to estimate the prevalence of ADHD symptoms among children in day treatment centres (N = 162) and residential treatment centres (N = 195) in Holland. Although further research is needed, the study supports the suggestion that such ratings can add to reliable diagnostic outcomes when assessing the behavioural symptoms of ADHD in children in the centres. It is estimated that nearly a fifth of the children in such centres exhibit the symptoms of ADHD in the judgement of professional care workers. Model testing using confirmatory factor analysis favours a dimensional behavioural model that comprises all the three constitutional symptom dimensions of ADHD (inattention, hyperactivity and impulsivity) instead of the two-factor model as used in the DSM-IV (inattention and hyperactivity/impulsivity). However, the differences of fit between both models were only small and the hyperactivity and impulsivity factors were highly correlated. This suggests that, in practice, a two-factor model can also be appropriate. The issue of whether a two-factor or a three-factor model is more appropriate thus remains unsolved in this study.     

EEG correlates of methylphenidate response in ADHD: association with cognitive and behavioral measures.

The authors examined the association between EEG correlates of medication response and concomitant cognitive and behavioral changes among children with attention-deficit hyperactivity disorder (ADHD). Subjects were 36 children with ADHD, aged 8 to 12 years. EEG activity was recorded from nine active electrodes during placebo and medication conditions. Medication administration resulted in increased alpha activity in central and parietal regions during both the baseline and cognitive activation conditions. Children who were medication responders exhibited increased frontal beta activity whereas nonresponders showed decreased beta activity in the same region. Increased frontal beta activity was significantly correlated with medication-related improvement in performance on Conners'Continuous Performance Test and parent behavior ratings in attention and hyperactivity. Decreased right frontal theta activity was associated with improvements in parent-rated attention, but not in CPT performance. Stimulant medication increases beta activity in children with ADHD, particularly in frontal regions. Increased cortical arousal and activation in the frontal cortex is strongly associated with sustained attention and response inhibition and with parent-rated attention and hyperactivity/impulsivity.     

Thermal balneotherapy induces changes of the platelet serotonin transporter in healthy subjects

Although the beneficial effects of balneotherapy have been recognized since a long time, a few information is available on the biological mechanisms underlying them and the subjective feelings of increased well-being and mood. The links between the serotonin (5-HT) system and mood prompted us to investigate the 5-HT platelet transporter (SERT), which is considered a reliable, peripheral marker of the same structure present in presynaptic neurons, in 20 healthy volunteers before (t0) and 30 min after (t1) thermal balneotherapy with ozonized water of Montecatini spa, as compared with a similar group who underwent a bath in non-mineral water. The SERT was evaluated by means of the specific binding of (3)H-paroxetine ((3)H-Par) to platelet membranes. Equilibrium-saturation binding data, the maximal binding capacity (Bmax) and the dissociation constant (Kd), were obtained by means of the Scatchard analysis. The results showed that, while Bmax values did not change in both groups, the Kd values decreased significantly at t1 only in those subjects who bathed in ozonized water. The results of this study, while showing a decrease of the dissociation constant (Kd) which is the inverse of affinity constant, of (3)H-Par binding to SERT in all subjects after balneotherapy and not in those bathing in normal water, suggest that SERT modifications may be related to a specific effect of ozonized water and, perhaps, also to the increased sense of well-being.     

Role of the serotonin system in ADHD: treatment implications

A limited number of studies have considered whether the activity of serotonin (5-hydroxytryptamine [5-HT]) contributes to the problems experienced by youngsters with attention-deficit/hyperactivity disorder (ADHD). The aim of this article is to review this work and propose interpretations. Peripheral measures of 5-HT and its metabolite do not point to a widespread association with the diagnosis. However, separate consideration of the major domains of dysfunction (motor activity, inattention and impulsivity) support a more differentiated assessment. The marked innervation of motor regions of the brain by 5-HT projections and the clear involvement of 5-HT systems in the control of locomotion in animals suggests a likely node for dysfunction in ADHD. The few relevant studies do not show evidence of this, but more attention should be accorded to the issue. The situation is different for attention-related processes; here, there are deficiencies in perceptual sensitivity and the appropriate designation of saliency to stimulation. These are attributable, in part, to altered 5-HT activity. Marked and opposite changes of 5-HT responsivity are associated with behavioral and cognitive impulsivity. There is also a growing series of studies demonstrating preferential transmission of various genetic markers for 5-HT receptors that are expressed in ADHD. Currently, the heterogeneity of methods in this young discipline restricts the possibilities of definition of these markers and the types of ADHD in which they are expressed.     

Impulsivity is related to striatal dopamine transporter availability in healthy males

Impulsivity characterises various psychiatric disorders, particularly attention-deficit/hyperactivity disorder (ADHD). Evidence shows that ADHD symptoms are associated with dopamine dysfunction and alleviated with methylphenidate, a drug that reduces dopamine transporter availability. ADHD-like symptoms and impulsive traits are continuously distributed across the general population. Here, we aimed to investigate the dopaminergic basis of impulsivity and other ADHD-related traits in healthy individuals by studying the association of these traits with striatal dopamine transporter availability. Single-photon emission computed tomography with [¹²³I] FP-CIT was performed on 38 healthy males. Impulsivity was measured using the Barratt Impulsiveness Scale (BIS) and hyperactivity-impulsivity and inattention using the Adult ADHD Self-Report Scale (ASRS). We found that greater dopamine transporter availability was associated with higher BIS impulsivity but not with ADHD-related traits. The association with BIS was significant after accounting for individual differences in age and neuroticism. These results suggest that individual differences in the dopamine system may be a neural correlate of trait impulsivity in healthy individuals.     

Serotonin-lesion Myoclonic syndromes. I. Neurochemical profile and S-1 receptor binding

This paper and the following one describe the effects of L-5-hydroxytryptophan (5-HTP) (after 3 intracisternal injections of 5,7-dihydroxytryptamine (DHT], fenfluramine (FF), p-chloroamphetamine (PCA) and drug combinations on (i) brain regional amine concentration (HPLC with LEC) and serotonin S-1 receptor binding; and (ii) 'serotonergic' behaviors in the same adult rats. Serotonin (5-HT) neurotoxins produced significantly different regional profiles of 5-HT depletion. Multiple DHT injections caused a 90-100% depletion of 5-HT concurrently in neocortex, hippocampus, striatum, septum/accumbens, pons, cerebellum, and cervical cord. Only PCA significantly depleted midbrain. Drug combinations with DHT resembled DHT alone rather than additive depletions, except for PCA + DHT, which produced a hybrid pattern of depletion. The S-1 binding assay, using cold 5-HT to displace [3H]5-HT, was performed with and without ascorbate, EDTA, CaCl2, and pargyline. Without ascorbate, binding was specific, saturable, region-dependent, and non-linear with high (Kd 1-3 nM) and low affinity (10-20 nM) components but no cooperativity (0.8 less than nH less than 1.0). Bmax and Kd did not differ significantly between vehicle- and drug-treated animals in neocortex, hippocampus, striatum, thalamus, hypothalamus, midbrain, pons, medulla, cervical cord, cerebellum, or septum/accumbens two weeks after lesioning, while the assay did detect a 60% reduction in Bmax induced by ascorbic acid (1 mM). The effects of assay conditions exceeded the changes sometimes reported in S-1 receptor Bmax after 5-HT lesions.     

Serotonergic measures in the brains of suicide victims: 5-HT2 binding sites in the frontal cortex of suicide victims and control subjects

The authors determined serotonin2 (5-HT2) binding in the frontal cortex of 32 suicide victims and 37 subjects who died from nonpsychiatric causes. The maximum number of binding sites (Bmax) and the affinity (Kd) were significantly higher in subjects who had committed suicide than in control subjects. However, there was no difference in Kd between these two groups after the influence of age, race, sex, and postmortem delay was covaried. The Bmax of subjects who had committed violent suicide was significantly greater than that of control subjects.