Measurement of Serotonin Turnover Rate in Rat Dorsal Raphe Nucleus by In Vivo Electrochemistry

5-Hydroxytryptamine (5-HT; serotonin) turnover rate in dorsal raphe nucleus of the urethane-anesthetized rat was estimated by using the in vivo electrochemical detector to measure the decay of extraneuronal 5-hydroxyindole acetic acid (5-HIAA) after monoamine oxidase inhibition. Carbon paste electrodes were scanned by semiderivative voltammetry and revealed two peaks: one at +0.15 V and the other at +0.25 V. The higher potential peak is composed primarily of the 5-HT metabolite 5-HIAA. After administration of pargyline, 75 mg/kg i.p., this peak declined exponentially. Regression analysis of these data by an exponential decay model yielded the fractional rate constant 0.82 +/- 0.06 h-1 (mean +/- SEM). This rate constant of 5-HIAA disappearance measured by in vivo electrochemistry is identical to the rate constant found by others measuring 5-HIAA disappearance by direct tissue assay methods. In animals not treated with pargyline, tissue 5-HIAA concentrations in the dorsal raphe nucleus were measured by HPLC with electrochemical detection. The average 5-HT turnover rate calculated as the product of the fractional rate constant and steady-state tissue 5-HIAA concentration was 12.6 nmol/g/h. These results demonstrate that electrochemical detection of extraneuronal 5-HIAA combined with monoamine oxidase inhibition can be used to measure neurotransmitter turnover in vivo in a discrete brain region.     

Altered serotonin and norepinephrine metabolism in rat dorsal raphe nucleus after drug-induced hypertension

The effect of drug-induced hypertension on neurotransmitter release from dorsal raphe nucleus was studied by in vivo electrochemical electrodes in urethane anesthetized male Sprague-Dawley rats. Carbon paste electrodes were stereotaxically placed into dorsal raphe nucleus and neurotransmitter release was estimated electrochemically. Blood pressure was recorded from a femoral arterial catheter. Voltammograms taken from dorsal raphe nucleus showed two distinct peaks corresponding to norepinephrine and 5-hydroxyindole acetic acid (5-HIAA). After basal blood pressure and neurotransmitter release were monitored for 30 min, blood pressure was raised 50 mmHg by continuous intravenous infusion of L-phenylephrine hydrochloride. Drug infusion was discontinued after 50 min, but blood pressure and neurotransmitter release were measured for an additional 2 hr. Results showed that the 5-HIAA response increased immediately after the initiation of hypertension and remained elevated. By contrast, norepinephrine release initially decreased, then returned to the basal level and then rose in parallel with 5-HIAA to a level above baseline as drug-induced hypertension was discontinued. The same experimental protocol was used to study the electrochemical response to drug-induced hypotension. Blood pressure was lowered 20 mmHg by intravenous infusion of sodium nitroprusside dihydrate. During hypotension, no changes were seen in either transmitter response. However, as reflex hypertension appeared following discontinuation of the sodium nitroprusside infusion, the 5-HIAA response increased and the norepinephrine response decreased. These results show that drug-induced and reflex hypertension reduce norepinephrine release and increase serotonin turnover in dorsal raphe nucleus in anesthetized normotensive rats. These reciprocal changes appear to be a part of the neural response to hypertension.     

Differential pulse voltammetry: focus on the measurement of 5-hydroxyindole compounds and uric acid in the brain

The electrochemical signal obtained at the +300 mV potential (peak 3) in rats implanted for more than one week could be dependent upon extracellular fraction of 5-hydroxyindolacetic acid (5-HIAA) since a single injection of Pargyline is sufficient to suppress it in caudate and raphe dorsalis nuclei. In contrast, in rats implanted for less than one week, this signal could be dependent upon extracellular fractions of 5-HIAA and uric acid since consecutive injections of Pargyline and Allopurinol are necessary to suppress it.     

In Vivo Electrochemical Monitoring of Serotonin in Spinal Dorsal Horn with Nafion-Coated Multi-Carbon Fiber Electrodes

Abstract: Biosensors sensitive for in vivo monitoring of serotonin (5-HT) in the CNS by differential normal pulse voltammetry were constructed by coating treated multi-carbon fiber electrodes (mCFEs) with Nafion (N-mCFE). In vitro sensitivities of mCFE and N-mCFE were compared in solutions ranging from 5 n M to 20 µ M of uric acid (UA), 5-hydroxyindoleacetic acid (5-HIAA), and 5-HT. The mCFEs were three to seven times less sensitive for 5-HIAA or UA than for 5-HT. Nafion treatment dramatically decreased sensitivity for 5-HIAA and UA of N-mCFEs (∼10³ times), whereas it remained in the nanomolar range for 5-HT. In vivo, in the dorsal horn of the lumbar spinal cord of anesthetized rats, the monoamine oxidase inhibitor clorgyline (10 mg/kg i.p.) produced a reduction (55 ± 3% at 180 min) of peak 3 of oxidation current (characteristic of 5-hydroxyindoles) monitored with mCFEs, but with N-mCFEs (in this latter case the peak was termed 3N) peak 3N increased to 135 ± 5% at 180 min. The 5-HT release-inducer p -chloroamphetamine (PCA; 6 mg/kg i.p.) induced a slight (12 ± 3% at 150 min) decrease in peak 3 measured with mCFEs, whereas with N-mCFEs PCA induced a rapid increase of peak 3N (137 ± 6% at 90 min). The xanthine oxidase inhibitor allopurinol (10 mg/kg i.p.) produced a decrease (30 ± 3% at 180 min) in peak 3 (mCFEs), but peak 3N (N-mCFEs) was not affected (106% at 180 min). After pretreatment with allopurinol, PCA also produced an increase (135 ± 6% at 90 min) in peak 3N. These in vitro and in vivo data provide evidence for a highly preferential detection of 5-HT versus 5-HIAA and UA by N-mCFEs, which could be used to follow the extracellular 5-HT concentration within very discrete structures throughout the CNS.     

Effect of Probe Size on the Concentration of Brain Extracellular Uric Acid Monitored with Carbon Paste Electrodes

Abstract: We have investigated further the anomalously high concentration of brain extracellular uric acid detected with in vivo sampling probes reported recently. The contribution by uric acid and 5-hydroxyindoleacetic acid (5-HIAA) to peak 2 recorded in rat striatum with chronically implanted carbon paste electrodes (CPEs) of different sizes was estimated by comparing peak current densities and the effect of the monoamine oxidase inhibitor pargyline. The concentration of uric acid in the extracellular fluid was some 50 times greater for 320-μm-diameter CPEs than for 160-μm-diameter electrodes, where the urate level was estimated at ∼1 μ M. The concentration of 5-HIAA was similar for 320-, 260-, and 160-μm-diameter CPEs. These data provide an explanation for the previously observed differences in 5-HIAA/urate ratios re corded with 320-μm-diameter CPEs and smaller carbon fibre electrodes. The results also indicate that chronically implanted sampling probes of diameter >160 μm perturb the surrounding tissue, which produces uric acid by a mechanism yet unknown, although preliminary histological data suggest that glial cells may be involved.     

Factors affecting in vivo electrochemistry: electrode-tissue interaction and the ascorbate amplification effect

While in vivo electrochemistry has been shown to be useful for discovering new neurophysiological phenomena, there is still considerable controversy about the identity of the compounds being measured and the concentration of those compounds in extracellular fluid in brain. We have found that carbon paste electrodes undergo changes in sensitivity and specificity for dopamine and other compounds after being implanted in brain. We have also examined the effect of ascorbate on the selective enhancement of catecholamine peaks to provide an explanation for the apparently very high concentrations of dopamine measured in the extracellular fluid space. After temporary brain implantation (20 min), carbon paste electrodes tested in vitro showed increased sensitivity and lower oxidation potentials for dopamine, norepinephrine and serotonin. These brain-treated electrodes also detected 3,4-dihydroxyphenylacetic acid (DOPAC) as a distinct peak at +0.16 V, although the electrode sensitivity for DOPAC was some 25 times lower than that for dopamine. Brain treatment did not alter electrode sensitivity or oxidation potential for 5-HIAA. The oxidation current for ascorbic acid when processed as the semiderivative showed no distinct peak in the potential range -0.2 to +0.4V for either untreated or brain-treated electrodes. However ascorbic acid amplified the electrochemical peaks of catechols in direct proportion to the ratio of the concentration of ascorbate to the concentration of the catechol. In the physiologic concentration range of 300 microM ascorbate, the electrochemical signal for 1 microM dopamine was amplified 4250%. While ascorbate amplification improves detectability of dopamine and norepinephrine, it also introduces ambiguity since changing catechol concentrations cannot be distinguished from changing ascorbate concentrations.     

Histochemical studies on the morphology of the golgi apparatus and its relationship to catecholamine biosynthesis in the locus coeruleus of the rat

Summary Detailed histochemical studies have been performed on the morphology of the Golgi apparatus (GA) by application of the thiamine pyrophosphatase (TPPase) method (Novikoff and Goldfischer, 1961) to the neurons of the locus coeruleus (LC) of normal and catecholamine biosynthesis inhibitors (fusaric acid and D, L--methyl-p-tyrosine methylester HCl) given adult healthy male Wistar strain rats. The neurons were classified into five categories on the basis of the morphology of the Golgi apparatus. The number of cells in individual categories was counted to evaluate the percentage of each category in the whole nucleus.The majority of cells belongs to Types II, III, and IV whose GA goes through cyclic activity, but the remaining neurons belong to Types I and V which may have a strong tendency to be different from the former in character. The latter neurons correspond formally with Types I and V of the rabbit LC, but they do not respond to the drugs administered. The rat LC is very similar to the dorsal vagal nucleus of the rabbit in regard to the dominant category. The present results indicate that the majority of the rat LC neurons may work vigorously and they may be motor neurons.Administration of the drugs caused reduction of TPPase activity, augmentation of disintegration and the budding-off process of the GA of Type IV, a decrease in the percentage of Type IV and an increase in that of Type II. Administration of 100 mg/kg fusaric acid caused maximal morphological change of the GA at the 90th minute; however, administration of 200 mg/kg fusaric acid showed more marked change of the GA, having two peaks and two valleys. The GA revealed much more intense reaction to D,L--methyl-p-tyrosine methylester HCl than to fusaric acid. The present results indicate that tyrosine hydroxylase may be the rate-limiting enzyme in the catecholamine biosynthesis.These noticeable changes of GA caused by administration of the drugs were completely restricted to the neurons of LC and the neurons of the mesencephalic nucleus of the trigeminal nerve did not show any morphological changes of the GA. These results strongly suggest that the GA of the rat LC neurons may have ability to synthesize catecholamine whereas the GA of the rat mesencephalic nucleus of the trigeminal nerve may be completely devoid of this ability and that the role of the GA may be different depending on the anatomical regions.     

尿酸在聚6-甲基香豆素修饰的玻碳电极上的电化学行为

制备了聚6-甲基香豆素修饰玻碳电极,研究了尿酸(UA)在该修饰电极上的电化学行为。实验结果表明:在pH=5.0的磷酸盐缓冲溶液中,扫描速率为50mV/s时,尿酸在修饰电极上于0.352V处产生一个灵敏的氧化峰,在0.278V处有一弱的还原峰。经线性扫描伏安法测定,氧化峰电流与尿酸浓度在2.5×10-6～1.0×10-5mol/L范围内表现出良好的线性关系,检出限为1.0×10-6mol/L。将修饰电极在常温下放置50d及将体系温度升高到75℃时,修饰电极对尿酸的响应电流大体不变,结果满意。     

Serotonin turnover in individual brain nuclei: evaluation of three methods using liquid chromatography with electrochemical detection

Liquid chromatography with electrochemical detection and brain microdissection techniques were used to evaluate three methods of studying serotonin turnover in 10 individual brain nuclei. The increase in serotonin (5-HT) and decline in 5-hydroxyindole acetic acid (5-HIAA) after administration of the monoamine oxidase inhibitor, pargyline, as well as the accumulation of 5-hydroxytryptophan (5-HTP) after the L-amino acid decarboxylase inhibitor, m-hydroxybenzylhydrazine, were measured. Serotonin accumulation and 5-HIAA decline could be detected in the n. caudatus, globus pallidus, cortical amygdala, n. interstitialis striae terminalis, n. preopticus medialis, and n. dorsomedialis. Only serotonin accumulation could be accurately assessed in the n. ventromedialis, n. arcuatus, and median eminence. The pattern of increase of serotonin after pargyline varied in different nuclei. There was a linear increase of serotonin over 90 minutes in the caudate, globus pallidus, and ventromedial nucleus and over 60 minutes in the n. preopticus medialis, and cortical amygdala. This contrasted with a maximal increase at 30 minutes in the other nuclei. However, 5-HIAA decline tended to be greatest after 30 minutes in most nuclei. Increases in 5-HTP concentrations after decarboxylase inhibition were not reliably detected in these areas. These results indicate that two nonsteady state methods may be used to evaluate changes in serotonin turnover in selected individual, nonpooled hypothalamic and forebrain nuclei.     

The Involvement of Fusaric Acid in the Bulb-rot of Gladiolus

Tissue cultures of Gladiolus have been used successfully to determine host specific properties of fusaric acid, a phytotoxin produced by Fusarium oxysporum f. sp. gladioli (Mas.) Sny. et Hn. Ten Gladiolus genotypes, including three wild South African species, varying in resistance to Fusarium-rot . were differentiated based on the expressed insensitivity to fusaric acid. Shoots and callus cultures were challenged in vitro with various concentrations of fusaric acid. The ion-release caused by the toxin was measured with callus and intact cormels. In all above mentioned bioassays resistant and susceptible genotypes could be generally discriminated. However, only two of the developed bioassays, the shoot assay and the ion-release with intact cormels, gave significantly coinciding results with the Fusarium-resistance assessed in a greenhouse experiment. When using callus tissue in the assays, the obtained answer correlated less with the Fusarium-resistance . It is concluded that a part of the Fusarium-resistance is based on fusaric acid insensitivity.     

Peripheral interleukin-6 administration increases extracellular concentrations of serotonin and the evoked release of serotonin in the rat striatum

Previous studies have indicated that peripheral administration of interleukin-6 (IL-6) increases brain concentrations of tryptophan and 5-hydroxyindoleacetic acid (5-HIAA), the major catabolite of serotonin (5-HT). To determine whether these changes were related to increased synaptic release of 5-HT, we studied the responses to peripheral administration of IL-6 by in vivo microdialysis and in vivo amperometry. Intraperitoneal injection of recombinant IL-6 resulted in an elevation of microdialysate concentrations of 5-HT in the rat striatum. Also, amperometric measurements indicated that i.p. IL-6 enhanced the 5-HT-like signal obtained from the striatum following electrical stimulation of the dorsal raphe nucleus. These results indicate that the increases in brain concentrations of 5-HIAA observed in earlier studies indeed reflect increased synaptic release of 5-HT.     

Drug-Induced Changes in Blood Pressure Lead to Changes in Extracellular Concentrations of Epinephrine, Dihydroxyphenylacetic Acid, and 5-Hydroxyindoleacetic Acid in the Rostral Ventrolateral Medulla of the Rat

Neurochemical changes in the extracellular fluid of the rostral ventrolateral medulla (RVLM) were produced by changes in arterial blood pressure. Blood pressure was raised or lowered with systemic infusions of phenylephrine or nitroprusside and neurochemicals were recovered from RVLM by in vivo microdialysis. A dialysis probe 300 microns in diameter and 500 microns in length was stereotaxically implanted in the RVLM of the urethane-anesthetized rat. Sterile physiological Ringer's solution was perfused at a rate of 1.5 microliter/min. The perfusate was collected under ice-cold conditions every 15 min for the assay of epinephrine, dihydroxyphenylacetic acid (DOPAC), 5-hydroxyindoleacetic acid (5-HIAA), ascorbic acid, and uric acid. After stable baseline neurochemical concentrations were achieved, animals were infused with phenylephrine or nitroprusside intravenously to raise or lower the blood pressure. Increasing blood pressure 50 mm Hg above the baseline value by phenylephrine led to a significant reduction in heart rate and a reduction in extracellular epinephrine and DOPAC concentrations. The 5-HIAA concentration was increased during the hypertensive drug infusion. There were no changes in the concentrations of ascorbic acid or uric acid. Hypotension produced by nitroprusside (-20 mm Hg) led to neurochemical changes which were the reciprocal of those seen during hypertension. During hypotension, heart rate increased as did the extracellular fluid epinephrine concentration. The 5-HIAA concentration fell with hypotension and remained depressed following the nitroprusside infusion. Ascorbic acid and uric acid concentrations did not change during hypotension but ascorbic acid did increase after the nitroprusside infusion stopped. These data provide direct evidence that epinephrine release in RVLM is linked to changes in systemic blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of lithium oxybutyrate on the serotonin and 5-hydroxyindoleacetic acid content in the brain of rabbits

Single administration of lithium hydroxybutyrate (10 mg/kg) to rabbits decreased serotonin and 5-hydroxyindoleacetic acid (5-HIAA) content in the caudate nucleus. The drug administration for 8 days is accompanied by mediator accumulation in the cortex, caudate nucleus, tonsils, hypothalamus, thalamus, and midbrain with parallel reduction in 5-HIAA level in these structures. 15 days of lithium hydroxybutyrate administration lead to the increase of serotonin and 5-HIAA concentration, while 28 days of administration reduced the content of mediator and its metabolite.     

2-Guanidinoethanol increased dopamine release and 3,4-dihydroxyphenylacetic acid content,but not homovanillic acid content in the rat brain: Electroneurochemical and enzymological studies

The effects of 2-guanidinoethanol (GEt) on the release of monoamines and on the activity of their degrading enzymes were studied in order to investigate why 3,4-dihydroxyphenylacetic acid (DOPAC) increased to a much greater extent than homovanillic acid (HVA) after GEt injection into rat brain. In differential pulse voltammograms recorded using an electrochemically treated carbon fiber electrode, two distinct oxidation peaks, one at 130mV (DOPAC peak) and the other at 300 mV (5-hydroxyindoleacetic acid (5-HIAA) peak), were observed. In the hippocampus, the DOPAC peak increased markedly compared to the peak height recorded prior to the intracerebroventricular injection of GEt (6mol). Although the DOPAC peak height increased to 350% 4 hours after GEt injection, the 5-HIAA peak showed no change. In the striatum, the DOPAC peak increased to 150% 3 hours after GEt injection. Serial changes in the extracellular levels of DOPAC, HVA, and 5-HIAA were monitored in the striatum after GEt injection, using an in vivo brain micro-dialysis technique. Although the DOPAC levels strated to increase 80 minutes after GEt injection, HVA and 5-HIAA levels showed no change. On the other hand, monoamineoxidase, which metabolizes dopamine to DOPAC, was not activated and catechol-0-methyltransferase, which metabolizes DOPAC to HVA, were not inhibited by 5 mM of GEt in vitro. These data suggested that GEt increased the release of dopamine, but not of serotonin, and that GEt might restrict the DOPAC transport system.     

In vivo release of endogenous dopamine, 5-hydroxytryptamine and some of their metabolites from rat caudate nucleus by phenylethylamine

The in vivo release of endogenous 3,4-dihydroxyphenylethylamine (DA) and its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 3-methoxytyramine (3-MT), and of 5-hydroxytryptamine (5-HT) and its metabolite, 5-hydroxyindoleacetic acid (5-HIAA), has been measured in the caudate nucleus of the anesthetized rat. A push-pull cannula was implanted into the brain, and the tissue perfused with artificial CSF or artificial CSF containing 5×10^–4 M phenylethylamine. The perfusate was collected and analyzed for DA, 5-HT and their metabolites by high performance liquid chromatography with electrochemical detection (HPLC-ECD). DA was released by phenylethylamine at rates significantly greater than its basal rate. 3-MT and 5-HT were undetectable in perfusates collected under basal conditions, but could be detected readlly during phenylethylamine stimulation. DOPAC, HVA and 5-HIAA concentrations were not significantly affected by phenylethylamine. The results suggest (1) that phenylethylamine may exert its behavioural effects through increased release of both DA and 5-HT, and (2) that in vivo measurements of the acid metabolites alone may not be indicative of the release of the amines.Special Issue Dedicated to Dr. Abel Lajtha.     

Current concepts: II. Measurement of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in discrete brain nuclei using reverse phase liquid chromatography with electrochemical detection

A rapid and sensitive method has been outlined for the measurement of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) utilizing a weak cation-exchange resin and liquid chromatography with electrochemical detection. The sensitivity of the procedure allows measurement of the amine in punches of rat substantia nigra even after local injection of the neurotoxin 5,7-dihydroxytryptamine. Increases in 5-HT and decreases in 5-HIAA concentrations after pargyline, and selective increases in 5-HIAA concentrations after probenecid were detected in selected brain regions (nucleus accumbens, anterior striatum, substantia nigra). Thus, this procedure is sensitive enough to estimate 5-HT turnover in discrete nuclei of the rat brain.     

孤束核蛋白酪氨酸激酶对外周化学感受性反射呼吸作用的影响

本工作旨在观察脑干孤束核内蛋白酪氨酸激酶(protein tyrosine kinase,PTK)是否参与了外周化学感受性反射的呼吸反应调节。实验采用电生理和微量注射相结合的方法,以膈神经放电为观察指标,观察呼吸变化。通过吸入10％氧气(10％O2,90％N2)引导出外周化学感受性反射。在孤束核(nucleus tractus solitarius,NTS)处分别微量注射蛋白酪氨酸激酶的抑制剂,genistein和其非活动性抑制剂daidzein以及AMPA受体阻断剂CNQX,观察这些药物对外周化学感受性反射的影响。结果显示,吸入低氧混合气后,动物呼吸加深加快;在NTS处微量注射CNQX或genistein都会不同程度削弱外周化学感受性反射引起的通气反应,而微量注射daidzein后对反射没有影响。另外,在NTS处微量注射CNQx后再注射genistein,其削弱外周化学感受性反射的作用与单独微量注射CNQx或genistein基本相同,二者并无协同作用。结果提示,NTS处的蛋白酪氨酸激酶对外周化学感受性反射具有一定的调节作用,并且NTS处磷酸化修饰,AMPA受体可能是PTK发挥这种调节作用的途径之一。     

In Vivo Monitoring of Catecholaminergic Metabolism in the C1 Region of Rat Medulla Oblongata: A Comparative Study by Voltammetry and Intracerebral Microdialysis

In vivo voltammetry or microdialysis was used to monitor catecholaminergic metabolism in the C1 region of the ventrolateral medulla oblongata of anesthetized rats. In vivo voltammetry allowed the recording of a catechol oxidation current (CA.OC) peak in this region. This CA.OC was suppressed after inhibition of monoamine oxidase by pargyline or after inhibition of tyrosine hydroxylase by alpha-methyl-p-tyrosine and was markedly increased after blockade of dopamine-beta-hydroxylase by FLA 63. Similar results were found when intracerebral microdialysis coupled with HPLC and electrochemical detection was used to measure the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC) in the dialysates obtained from the C1 region: The changes in CA.OC and DOPAC concentration in the dialysates exhibited very similar kinetic characteristics in the three pharmacological experiments. These results support the involvement of DOPAC as a major component of the electrochemical signal recorded by voltammetry in the C1 group of adrenergic neurons.     

A rapid and simple method to determine the specific activities of serotonin, 5-hydroxyindoleacetic acid,and 5-hydroxytryptophan in brain by HPLC with electrochemical detection

The specific activities of 5-hydroxytryptophan (5-HTP), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) have been determined in the brain of rats by HPLC using electrochemical detection. The method allows, from a single sample, the simultaneous measurement of all three compounds and collection of each peak for radioactivity determinations. Five male Wistar rats were injected i.v. with 2.0 mCi/kg ofDl-5-hydroxy-[G-³H]tryptophan (2.6 Ci/mmol) and 30 min later the animals were killed by near freezing. Whole brains were removed and homogenized in an acid medium. The content of 5-HTP, 5-HT, and 5-HIAA were determined by HPLC. Each peak of interest was immediately collected after detection in scintillation vials by use of a small dead space detector (TL-9A, Bioanalytical Systems, Inc.). The amounts of radioactivity were determined and specific activities calculated from the results. A second chromatography system (TLC) was used to check the authenticity and purity of compounds separated by the HPLC.     

Effect of 3,4-Methylenedioxymethamphetamine on 3,4-Dihydroxyphenylalanine Accumulation in the Striatum and Nucleus Accumbens

The effect of the racemic mixture of 3,4-methylenedioxymethamphetamine (MDMA) on the synthesis of dopamine in the terminals of nigrostriatal and mesolimbic neurons was estimated by measuring the accumulation of 3,4-dihydroxyphenylalanine (DOPA) in the striatum and nucleus accumbens 30 min following the administration of the L-aromatic amino acid decarboxylase inhibitor, 3-hydroxybenzylhydrazine. MDMA produced an increase in DOPA accumulation in the striatum which was greater in magnitude and longer in duration than that in the nucleus accumbens. Although the concentrations of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in both the striatum and nucleus accumbens were reduced 3 h following an injection of MDMA (20 mg/kg), 5-HT and 5-HIAA concentrations were significantly reduced only in the striatum 7 days after the administration of MDMA. Pretreatment with a 5-HT2 antagonist, ketanserin, significantly attenuated the reduction in 5-HT concentration in the striatum 3 h following MDMA administration and completely blocked 5-HT depletion at 7 days post administration. Moreover, ketanserin completely blocked MDMA-induced DOPA accumulation in the striatum. The results obtained in these studies suggest that MDMA activates nigrostriatal dopaminergic pathways via 5-HT2 receptors. In addition, these data are supportive of the hypothesis that dopamine plays a role in MDMA-induced 5-HT depletion.