Effect of the attention deficit/hyperactivity disorder drug atomoxetine on extracellular concentrations of norepinephrine and dopamine in several brain regions of the rat

Atomoxetine is a selective inhibitor of norepinephrine transporters and is currently being used in the pharmacotherapy of attention deficit/hyperactivity disorder (ADHD). We have previously shown that atomoxetine increased extracellular (EX) concentrations of norepinephrine and dopamine in prefrontal cortex, but unlike the psychostimulant methylphenidate, did not alter dopamine(EX) in nucleus accumbens or striatum. Using the in vivo microdialysis technique in rat, we investigated the effects of atomoxetine on norepinephrine(EX) and dopamine(EX) concentrations in several other brain regions and also evaluated the role of inhibitory autoreceptors on atomoxetine-induced increases of norepinephrine(EX) concentrations. Atomoxetine (3mg/kg i.p.) increased norepinephrine(EX) robustly in prefrontal cortex, occipital cortex, lateral hypothalamus, dorsal hippocampus and cerebellum, suggesting that norepinephrine(EX) is increased throughout the brain by atomoxetine. In lateral hypothalamus and occipital cortex where dopamine(EX) was quantifiable, atomoxetine did not increase dopamine(EX) concentrations, in contrast to parallel increases of norepinephrine(EX) and dopamine(EX) in prefrontal cortex, indicating a unique effect in prefrontal cortex. Administration of the alpha(2)-adrenergic antagonist idazoxan 1h after atomoxetine resulted in increases in prefrontal cortical norepinephrine efflux greater than either compound alone, indicating an attenuating effect of the adrenergic autoreceptors on norepinephrine efflux.     

In vivo release of newly synthesized catecholamines from the hypothalamus by amphetamine

Summary The posterior hypothalamus of cats immobilized with gallamine was superfused through a push-pull cannula with artificial cerebrospinal fluid. Addition of³H-tyrosine into the superfusing fluid led to synthesis of³H-catecholamines which were released spontaneously. Separation of the³H-catecholamines by column chromatography or their acetylation and separation by paper chromatography revealed that both³H-noradrenaline and³H-dopamine were released. In most experiments³H-noradrenaline represented about 10 to 25% of total³H-catecholamines. Superfusion of the hypothalamus with amphetamine (1×10^–5 M) enhanced the release of total³H-catecholamines, the release of³H-noradrenaline being relatively more enhanced than that of³H-dopamine. Determination of the readioactive compounds in the hypothalamus at the end of the experiments showed that total³H-catecholamines represented 3% of³H-tyrosine. About 15% of the total³H-catecholamines were due to³H-noradrenaline and 85% to³H-dopamine.     

Low dopamine transporter occupancy by methylphenidate as a possible reason for reduced treatment effectiveness in ADHD patients with cocaine dependence

Methylphenidate (MPH) occupies brain striatal dopamine transporters (DATs) and is an effective treatment for attention deficit hyperactivity disorder (ADHD). However, patients with ADHD and comorbid cocaine dependence do not benefit significantly from treatment with MPH. To better understand the neurobiology of this phenomenon, we examined DAT availability and the effects of MPH treatment on DAT occupancy in ADHD patients with and without cocaine dependence. ADHD patients without a comorbid substance use disorder (N=16) and ADHD patients with comorbid cocaine dependence (N=8) were imaged at baseline and after two weeks MPH treatment using single photon emission computed tomography (SPECT) with the DAT tracer [¹²³I]FP-CIT. Changes in ADHD symptoms were measured with the ADHD symptom rating scale (ASRS). At baseline, we observed lower striatal DAT availability in ADHD patients with cocaine dependence. Following fixed MPH treatment, MPH occupied significantly less striatal DATs in cocaine-dependent than in non-cocaine dependent ADHD patients. There were no significant correlations between baseline DAT availability or DAT occupancy by MPH and ADHD symptom improvement. However, we did find significant correlations between DAT occupancy by MPH and decreases in impulsivity scores and years of cocaine use. These preliminary findings suggest that low DAT occupancy is not the reason why ADHD patients with cocaine dependence do not benefit from MPH treatment. It also suggests that higher dosages of MPH in these patients are probably not the solution and that medications directed at other pharmacological targets should be considered in these comorbid ADHD patients.This trial is registered at the Dutch Trial Register, www.trialregister.nl, under Trial ID number NTR3127.     

Influence of morphine and naloxone on the release of noradrenaline from rat brain cortex slices

Summary In slices of rat brain cortex preincubated with (–)-³H-noradrenaline, the influence of morphine and naloxone on the efflux of tritium was investigated. The spontaneous outflow of tritium was not changed by 10^–7–10^–5 M morphine and by 10^–6–10^–4 M naloxone, but was accelerated by 10^–4 M morphine. Electrical field stimulation augmented tritium outflow. The overflow evoked per ppulse decreased as the frequency of stimulation was increased from 0.3 to 3 Hz, but remained approximately constant when it was further increased to 10 Hz. At frequencies of 0.3, 1, and 3 Hz, but not at 10 Hz, morphine in concentrations of 10^–7–10^–5 M depressed the stimulation-induced overflow of tritium. 10^–4 M morphine did not influence the overflow induced by stimulation at 0.3 and 1 Hz and increased that evoked by stimulation at 10 Hz. Naloxone (10^–6–10^–4 M) did not change the response to stimulation. In the presence of 10^–4 M naloxone, 10^–6 M morphine did not diminish, and 10^–5 M morphine even enhanced the stimulation-induced overflow of tritium. The inhibitory effect of 10^–6 M morphine was not reduced, after tyrosine hydroxylase had been blocked by -methyltyrosine-methylester. It is concluded that morphine through an action on specific opiate receptors inhibits the release of transmitter from cerebrocortical noradrenergic neurones evoked by nerve impulses. By an action unrelated to opiate receptors, morphine at high concentrations increases the stimulation-induced overflow of noradrenaline, presumably by inhibiting its re-uptake into nerve endings.     

Effects of dopamine D4 receptor-selective antagonists on motor hyperactivity in rats with neonatal 6-hydroxydopamine lesions

RATIONALE: Dopamine D4 receptor gene polymorphism has been repeatedly associated with attention deficit hyperactivity disorder (ADHD) and related personality traits. We recently reported that motor hyperactivity in an animal model of ADHD was dose-dependently reversed by CP-293,019, a D4 receptor-selective antagonist. However, behavioral effects of this agent may not be attributed exclusively to D4 receptor blockade, since it interacts with other sites including serotonin receptors. OBJECTIVES: To test further the hypothesis that D4 receptor blockade can reduce motor hyperactivity, behavioral effects of three chemically and pharmacologically dissimilar D4 antagonists were compared to that of ketanserin, a serotonin 5-HT(2A/2C) antagonist. METHODS: Selective dopamine lesions were made in male rats at postnatal day (PD) 5 with intracisternal 6-hydroxydopamine (100 microg) after desipramine pretreatment (25 mg/kg, SC) to protect noradrenergic neurons. Effects of D4 receptor-selective antagonists and ketanserin on lesion-induced motor hyperactivity were examined during the periadolescent period (postnatal days 23-26) with an infrared photobeam activity system. RESULTS: The D4 antagonists L-745,870 and U-101,958 dose-dependently inhibited motor hyperactivity in rats with neonatal lesions, whereas S-18126 lacked this effect at doses up to 30 mg/kg. None of these drugs affected motor behavior in sham control rats. In contrast, ketanserin produced apparent sedative effects in both lesioned and intact control rats without normalizing hyperactivity. CONCLUSIONS: Motor hyperactivity in this ADHD model was selectively antagonized by three of four dopamine D4 receptor antagonists evaluated, encouraging clinical assessment of D4 antagonists in patients with ADHD.     

Effect of sympathomimetic amines on the synaptosomal transport of noradrenaline,dopamine and 5-hydroxytryptamine

The interaction of sympathomimetic amines with the transport of ³H-noradrenaline (³H-NE), ³H-dopamine (³H-DA) and ³H-5-hydroxytriptamine (³H-5-HT) were investigated in rat hypothalamic (³H-NE) and striatal (³H-DA and ³H-5-HT) synaptosomes. Modifications in the phenylethylamine structure led to changes in activity towards biogenic amine uptake and release: (a) the introduction of a β-OH group led to compounds less active in inhibiting uptake and stimulating release of ³H-NE, ³H-DA and ³H-5-HT, with the exception of ³H-NE release which was stimulated more by unlabeled 1-NE than by DA; (b) the introduction of phenolic -OH groups always led to compounds which were stronger uptake inhibitors and releasers of the three biogenic amines; (c) the α-methylation increased the potency towards uptake inhibition and release stimulation, with the exception of ³H-NE release: in fact, the releasing activity of phenylethylamine was suppressed by α-methylation; (d) the introduction of a -Cl group in the para position selectively potentiated the effects on ³H-5-HT uptake and release and generally depressed those on catecholamine transport.     

Influence of fentanyl,levorphanol and pethidine on the release of noradrenaline from rat brain cortex slices

Summary In slices of rat brain cortex preincubated with (–)-³H-noradrenaline, the influence of fentanyl, levorphanol and pethidine on the efflux of tritium was investigated. The spontaneous outflow of tritium was not changed by low, and was accelerated by high concentrations of the drugs. The overflow of tritium evoked by electrical stimulation at 3 Hz was diminished by 10^–8–10^–7 M fentanyl and by 10^–7–10^–6 M levorphanol, but was augmented by 10^–5 M levorphanol. Naloxone prevented the inhibitory effect of fentanyl and levorphanol. In contrast to fentanyl and levorphanol, pethidine did not decrease, but at concentrations of 10^–6–10^–5 M greatly increased the stimulation-induced overflow of tritium. However, the increase was abolished, and the stimulation-evoked overflow slightly reduced, after the re-uptake of noradrenaline had been blocked by cocaine. It is concluded that fentanyl, levorphanol and pethidine share with morphine the ability to inhibit the release of transmitter from cerebrocortical noradrenaline neurones evoked by nerve impulses.     

Cyclic AMP facilitates the electrically evoked release of radiolabelled noradrenaline,dopamine and 5-hydroxytryptamine from rat brain slices

Summary The adenylate cyclase activator forskolin as well as 8-bromo-cyclic AMP enhanced the electrically evoked release of³H-noradrenaline and³H-5-hydroxytryptamine from superfused rat neocortical slices and that of³H-dopamine from neostriatal slices with comparable EC₅₀'s of about 0.5 and 50 M, respectively, without affecting spontaneous tritium efflux. The phosphodiesterase inhibitor ZK 62771 (3–100 M) also enhanced³H-noradrenaline and³H-dopamine release but slightly reduced³H-5-hydroxytryptamine release. However, this drug profoundly enhanced spontaneous tritium release in the latter case. The facilitatory effect of forskolin (0.3 M) on the release of the amine neurotransmitters was potentiated in the presence of ZK 62771 (30 M). Therefore, cyclic AMP appears to exert a general facilitatory effect on the release of these biogenic amines from central nerve terminals.     

国内盐酸哌甲酯控释片治疗儿童注意缺陷多动障碍的Meta分析

目的应用Meta分析评价国内盐酸哌甲酯控释片(专注达)与传统药物治疗18岁以下儿童注意缺陷多动障碍(ADHD)的临床疗效。方法通过中国学术期刊网全文数据库(CNKI,1979-2008)、中国生物医学文献数据库(CBMdisc,1978-2008)和国家科技图书文献中心等,检索国内有关盐酸哌甲酯控释片治疗儿童ADHD的随机临床对照试验,由两位评价者进行质量评价,并对符合纳入标准的临床试验研究进行Meta分析。结果共查阅到国内相关临床研究13篇,最终进入评价的临床研究4篇,患儿174例。合并分析表明,盐酸哌甲酯控释片与传统治疗药物比较,可显著降低患儿ADHD-RS量表的评分(P<0.01),平均减分值之差(95%CI)为[-7.85(-9.07,-6.64)]。结论从现有的临床证据看,盐酸哌甲酯控释片在改善18岁以下ADHD患儿的核心症状方面疗效显著。但由于样本数较少,该药与传统治疗药物的疗效对比仍需大规模、高质量、随访结局统一的临床试验进一步验证。     

Effect of chronic clonidine treatment on the turnover of noradrenaline and dopamine in various regions of the rat brain

Summary The turnover of noradrenaline (NA) and dopamine (DA) was estimated in various rat brain regions by measuring the depletion of the amines after inhibition of their biosynthesis by -methyltyrosine. Acute treatment with clonidine (0.1 mg/kg) reduced NA turnover in the brain stem, hypothalamus and rest of the brain but had no effect on DA turnover in the corpus striatum and rest of the brain. After chronic clonidine treatment (0.1 mg/kg, twice daily for 15 days), NA turnover was not affected by an additional injection of clonidine in the brain stem or in the hypothalamus but was still markedly reduced in the rest of the brain. In addition, DA turnover was reduced in the corpus striatum and rest of the brain, an effect which was also observed after a single injection of a high dose of clonidine (1 mg/kg). These findings suggest that a chronic administration of clonidine may cause regionally differential changes in the sensitivity of central NA receptors.     

Differential effects of amphetamine isomers on dopamine release in the rat striatum and nucleus accumbens core

Rationale Current medications for attention-deficit/hyperactivity disorder (ADHD) include some single isomer compounds [dextroamphetamine (d-amphetamine, dexedrine) and dexmethylphenidate (Focalin)] and some racemic compounds [methylphenidate and mixed-salts amphetamine (Adderall)]. Adderall, which contains approximately 25% l-amphetamine, has been successfully marketed as a first-line medication for ADHD. Although different clinical effects have been observed for d-amphetamine, Adderall, and benzedrine; potential psychopharmacological differences on the level of neurotransmission between d-amphetamine and l-amphetamine have not been well characterized.Objectives To evaluate potential differences in the isomers, we used the technique of high-speed chronoamperometry with Nafion-coated single carbon-fiber microelectrodes to measure amphetamine-induced release of dopamine (DA) in the striatum and nucleus accumbens core of anesthetized male Fischer 344 rats. Amphetamine solutions were locally applied by pressure ejection using micropipettes.Results The presence of l-amphetamine in the d,l-amphetamine solutions did not cause increased release of DA but did change DA release kinetics. The d,l-amphetamine-evoked signals exhibited significantly faster rise times and shorter signal decay times. This difference was also observed in the nucleus accumbens core. When l-amphetamine was locally applied, DA release was not significantly different in amplitude, and it exhibited the same rapid kinetics of d,l-amphetamine.Conclusions These data support the hypothesis that amphetamine isomers have different effects on release of DA from nerve endings. It is possible that l-amphetamine may have unique actions on the DA transporter, which is required for the effects of amphetamine on DA release from nerve terminals.     

Mazindol and amphetamine as inhibitors of the uptake and releasers of 3H-dopamine by rat striatal synaptosomes

Summary The effects of mazindol, amphetamine and fentluramine on uptake and release of ³H-DA by synaptosomes were studied in different systems.In in vitro incubations of ³H-DA with synaptosomes isolated from the caudate nucleus of the rat, mazindol inhibited the uptake of the radioactivity more potently than did amphetamine.When the synaptosomes were isolated from the caudate nuclei of rats treated in vivo with either mazindol or amphetamine, the uptake of ³H-DA during in vitro incubation was lower with synaptosomes of amphetamine-treated rats than with those of mazindol-treated rats.When synaptosomes of untreated rats were prelabelled with ³H-DA and incubated in the presence of amphetamine or of mazindol, amphetamine caused a greater releaseoof radioactivity than did mazindol.Fenfluramine was without activity in all these systems.In spite of the quantitative differences, both amphetamine and mazindol appear to have similar effects on uptake and release of dopamine, and this may account for their analogous pharmacological profile.Supported by C.N.R. grant N. 75.00620.04.115.2380     

Effects of dopamine on veins in humans: comparison with noradrenaline and influence of age

Objective: To compare the venoconstricting effect of dopamine with that of noradrenaline and to investigate the influence of age on the responsiveness to dopamine in human subjects. Methods: In eight young and eight elderly male subjects, increasing doses of dopamine or noradrenaline were infused into a dorsal hand vein and its diameter was measured using a linear variable differential transformer. Results: There was no significant difference between the maximum venoconstriction (E_max) for dopamine and that for noradrenaline. The infusion rate to induce 50% of E_max (ED₅₀) for dopamine in the young and elderly subjects was 363 ng · min⁻¹ and 352 ng · min⁻¹, and the ED₅₀ for noradrenaline was 40.7 ng · min⁻¹ and 43.8 ng · min⁻¹, respectively. Neither in the E_max nor in the ED₅₀ for these drugs were there significant differences between the young and elderly subjects. Conclusion: The venoconstricting effect of dopamine is 5–20 times less than that of noradrenaline, and aging does not influence the responsiveness to dopamine and noradrenaline in human subjects. Received: 29 August 1997 / Accepted in revised form: 5 February 1998     

Effects of chronic amphetamine intoxication on the accumulation in the rat brain of labelled catecholamines synthesized from circulating tyrosine-14C and dopa-3H

Summary Accumulation of labelled noradrenaline (NA) and dopamine (DA) and endogenous levels of NA and DA in brain and heart were measured after infusion of tyrosine-¹⁴C and dopa-³H in rats. After a single dose of 20 mg/kg i.p. of dl-amphetamine sulphate a 50–65% decrease in the NA accumulation in the brain and heart was observed. The accumulation of DA was not changed. In rats treated chronically with amphetamine sulphate, 16–32 mg/kg i.p. twice daily for 4 weeks, both the brain and heart NA and brain DA levels were depleted by 40–70% at 20–24 h after amphetamine withdrawal. The accumulation of labelled NA and DA in these animals was not different from that of saline treated controls. When amphetamine sulphate, 20 mg/kg i.p., was given to chronically amphetamine treated rats there was a 45–60% decrease in the accumulation of labelled NA; labelled DA remained unchanged. It is concluded that the decrease in NA accumulation under acute and chronic amphetamine intoxication is most likely due to a preferential release of newly synthesized NA.The skilful technical assistance of Mrs Inger Jansson is gratefully acknowledged. A sample of 3-methoxytyrosine was generously supplied by AB Hässle, Mölndal, Sweden. The study was supported by the Swedish Medical Research Council (Project B71-14X-1017-07 and B71-27P-2627-03) and the Tri-Centennial Fund of the Bank of Sweden (Project Nr. 150).     

Differential effects of hypothermia on neuronal efflux,release and uptake of noradrenaline

Summary Isolated rabbit hearts were perfused at 34° (control), 24° or 12°C. The neuronal efflux of noradrenaline after perfusion with the amine for 1 h was depressed at 24° C (Q ₁₀ about 5) in the presence or absence of desipramine; at 12°C the efflux was below the limit of estimation. Moderate reduction of the temperature (24° C) decreased the removal of perfused noradrenaline to about 60% of the control value and caused a 1.7-fold increase of the output of noradrenaline evoked by sympathetic nerve stimulation. It is concluded that the extremely temperature-dependent efflux of noradrenaline across the axonal membrane is not part of the release of noradrenaline evoked by nerve stimulation.     

Differential effects of D- and L-amphetamine and methylphenidate on rat striatal dopamine biosynthesis

Methylphenidate and the optical isomers of amphetamine have differential effects, in vivo and in vitro on rat striatal synaptosomal dopamine (DA) biosynthesis. Whereas the systemic administration of D- or L-amphetamine produces a dose-dependent decrease in synaptosomal DA synthesis, with ED₅₀'s of 1.1 and 3.5 mg/kg, respectively, methylphenidate, at doses which produce comparable degrees of stereotyped behavior, has no effect on this measure of DA biosynthesis. In vitro, D- and L-amphetamine exhibit a three-fold difference in ED₅₀'s (1 × 10⁻⁶ M and 3 × 10⁻⁶ M, respectively) with respect to both activation of striatal synaptosomal DA synthesis, and to reversal of reserpine-induced inhibition of DA synthesis. In contrast, although in vitro methylphenidate can partially activate DA synthesis, it does not overcome the reserpine-induced inhibition. The data are discussed in terms of the possible differential mechanisms by which these two types of stimulants may exert their effects on stereotypy.     

Sequence variation in the human dopamine transporter gene in children with attention deficit hyperactivity disorder

The activity of the presynaptic dopamine (DA) transporter (DAT) is critical in mediating the magnitude and duration of dopaminergic signaling in the brain. Multiple genetic studies have found an association between attention deficit hyperactivity disorder (ADHD) and a variable number tandem repeat (VNTR) in the 3'-untranslated region (3'VNTR) of the hDAT gene (SLC6A3), however none of these studies examined the hDAT coding region for polymorphisms. Thus, we sought evidence of polymorphisms in hDAT, focusing on the coding region and splice junctions, utilizing genomic DNA from children diagnosed with ADHD. Two separate ADHD cohorts (N=70 and N=42) were screened and sampled for both status of the 3'VNTR and for common/novel genomic variants. We found evidence of increased DAT variation in African-American subjects as well as in predominantely hyperactive-impulsive probands. Cumulatively, multiple hDAT sequence variants were identified, including five novel variants, as well as one nonsynonymous single nucleotide polymorphism (SNP), converting Ala559 to Val (A559V). A559V was identified in two Caucasian male siblings with ADHD and both subjects were homozygous for the ADHD-associated, 10-repeat 3'VNTR allele. Interestingly, the A559V variant was previously identified in a subject with bipolar disorder [. Molecular Psychiatry 5, 275], a psychiatric disorder that has a significant number of overlapping symptoms with ADHD.