The 5-HT1A receptor agonist, 8-OH-DPAT, attenuates stress-induced anorexia in conjunction with the suppression of hypothalamic serotonin release in rats

The effect of the selective 5-HT_1A receptor agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) on stress-induced anorexia and serotonin (5-HT) release in the rat hypothalamus was studied with brain microdialysis. Subcutaneous injection of 8-OH-DPAT (1 mg/kg) significantly attenuated the immobilization-induced anorexia for 3 h, but had no effect during the following 9 h. Injection of 8-OH-DPAT itself had no effect on basal release of 5-HT, while it significantly blocked the immobilization-induced 5-HT release in the lateral hypothalamus. The results suggest that 8-OH-DPAT attenuated the stress-induced anorexia through the activation of 5-HT_1A autoreceptors in dorsal raphe nucleus.     

Characteristics of feeding induced by the serotonin agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT)

The effects of the putative serotonin agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) on food intake in freely-feeding and food deprived rats were examined. In freely-feeding rats, low doses of 8-OH-DPAT (15-60 micrograms/kg) significantly increased food intake without affecting drinking, grooming, rearing or locomotion. Higher drug doses (125-4000 micrograms/kg) also produced feeding and caused locomotor stimulation and serotonin-related stereotyped behaviour (i.e., forepaw padding, headweaving, wet dog shakes, flat body posture). When feeding and stereotypy were observed concurrently, response competition was evident and feeding behaviour was fragmented into numerous short eating bouts. As drug-induced stereotypy declined with time, this fragmented pattern of eating was succeeded by long bouts of eating which were similar to those observed at doses of 15-60 micrograms/kg 8-OH-DPAT. In 24 hr food deprived rats, low doses of 8-OH-DPAT had no effect on food intake. However, high doses of 8-OH-DPAT (250-4000 micrograms/kg) decreased feeding in food deprived animals, an effect which was probably secondary to the induction of stereotypy. It is proposed that the behavioural effects of 8-OH-DPAT may be explained by a dual effect on brain serotonergic mechanisms, which is dose dependent. Thus, low doses of the drug may preferentially activate inhibitory presynaptic serotonin receptors (autoreceptors), decrease serotonin metabolism and thereby increase feeding. In contrast, high doses of 8-OH-DPAT appear to stimulate postsynaptic serotonin receptors and thus produce stereotypy. Alternatively, it is possible that 8-OH-DPAT may elicit feeding by postsynaptic serotonin receptor blockade.     

Medial septal unit firing characteristics following injections of 8-OH-DPAT into the median raphe nucleus

Extracellular single-unit recording techniques were used to examine the firing characteristics of neurons in the medial septum/diagonal band of Broca complex (MS/1313) following injections of the 5-HT_1A agonist, 8-OH-DPAT, into the median raphe nucleus (MRN) of urethane-anesthetized rats. It had previously been shown that MRN injections of 8-OH-DPAT produce hippocampal theta rhythm. Injections of 8-OH-DPAT into the MRN produced a change in firing characteristics of MS/1313 neurons from an irregular discharge to a pattern of rhythmical bursting in synchrony with hippocampal theta rhythm. Cross-correlational and coherence analyses demonstrated that the rhythmical firing pattern of MS/1313 neurons strongly correlated with rhythmical fluctuations in the hippocampal EEG during periods of hippocampal theta produced by 8-OH-DPAT injections, but not during baseline conditions (i.e. hippocampal desynchronization). The results suggest that MRN control of the hippocampal EEG is modulated by the MS/DB. Serotonergic projections from the MRN to the MS/DB may normally act to inhibit the rhythmical bursting of MS/1313 neurons, thereby producing hippocampal desynchronization. Suppression of MRN 5-HT neurons by MRN injections of 8-OH-DPAT would disinhibit MS/1313 neurons, allowing them to burst rhythmically and thereby produce hippocampal theta rhythm.     

Suppression of conditioned avoidance by 8-OH-DPAT in the rat

Summary Rats were trained to perform an aversely motivated discriminative task in a shuttle-box. The conditioned avoidance response was selectively suppressed by 8-OH-DPAT in a dose-dependent manner (25–100 g·kg^–1). There were no statistically significant deficits in discriminative performance. The present results suggest antipsychotic-like properties of 8-OH-DPAT.     

Effects of acute treatment with 8-OH-DPAT and fluoxetine on aggressive behaviour in male song sparrows (Melospiza melodia morphna)

The role of serotonin in modulating male aggressive behaviour was investigated in male song sparrows, Melospiza melodia morphna, using two different serotonergic drugs, fluoxetine and 8-OH-DPAT. Fluoxetine is a selective serotonin reuptake inhibitor of the neuronal reuptake pump increasing synaptic concentrations of serotonin, and 8-OH-DPAT is a specific serotonin (5-HT1A) receptor agonist. The serotonergic control of aggression in passerines has not been previously investigated. We examined these behaviours within a controlled setting using a laboratory simulated territorial intrusion, with a hierarchical scale to quantify male-male aggressive behaviour. Utilizing this scale, we quantified the extent of male aggressive behaviour in two experiments. In experiment 1, song sparrows were given 100 micro l, s.c. injections of either fluoxetine (10 mg/kg) or 8-OH-DPAT (1 mg/kg). Experiment 2 was a dose-response study using three doses of 8-OH-DPAT (0.1, 1 and 10 mg/kg). In both studies, aggressive behaviour was measured 1 h after injection for 10 min in response to the presence of a novel male decoy combined with playback of conspecific song. Both drugs significantly reduced male aggressive behaviour, and 8-OH-DPAT did so in a dose-dependent manner. The effect of the two drugs upon general activity was also measured using infra-red perch hop detectors. Activity levels were not effected by either fluoxetine or 8-OH-DPAT at all of the respective doses, indicating that the reduction in aggressive behaviour was specific. These results demonstrate that, in a passerine species, the serotonergic system negatively regulates male-male aggressive behaviour. These results further demonstrate that aggression can be effectively studied in a laboratory setting and natural aggressive responses can be elicited using this method.     

Citalopram and 8-OH-DPAT attenuate nicotine-induced excitation of central noradrenaline neurons

Summary Previous electrophysiological studies have demonstrated that nicotine, intravenously administered, excites noradrenergic neurons in the locus coeruleus (LC) indirectly by releasing excitatory amino acids (EAA). In the present study the excitatory action of nicotine was inhibited by treatment with the selective 5-HT re-uptake inhibitor citalopram or the 5-HT_1A receptor agonist 8-OH-DPAT. It is proposed that the antagonism between nicotine and citalopram or 8-OH-DPAT reflects an interaction between endogenous EAA, e.g. glutamate, and 5-HT. The results may, on a cellular basis, explain the attributed effectiveness of drugs that facilitate serotonergic neurotransmission in promoting smoking cessation.     

In vivo modulation of acetylcholine in the nucleus accumbens of freely moving rats: I. Inhibition by serotonin

Microdialysis was used to characterize the effect of serotonergic input on cholinergic interneurons in the nucleus accumbens (NAC) of freely moving rats. Local infusion of 5-hydroxytryptamine (5-HT) or the serotonin reuptake blocker fluoxetine significantly decreased extracellular acetylcholine (ACh) in the NAC. This decrease in ACh was blocked by the 5-HT₁ (and β-adrenergic) antagonist propranolol. To test β-adrenergic effects, it was found that a β-adrenergic agonist isoproterenol had no measurable effect on extracellular ACh in the NAC. This suggests that 5-HT inhibits ACh interneurons via one of the 5-HT₁ receptor types. The 5-HT_1A agonist 8-OH-DPAT given systematically again decreased extracellular levels of ACh, and the effect was dose-dependent. The 5-HT_1A effect was probably exerted in the NAC, because local infusion of 8-OH-DPAT mimicked systemic injections. These microdialysis results are similar to in vitro studies which suggest an inhibitory impact of 5-HT on ACh release in basal ganglia slices and homogenates. The decrease in extracellular ACh as measured in vivo is apparently mediated, at least in part, through a 5-HT_1A receptor in the accumbens. Given the role of the NAC in behavior reinforcement, this 5HT-ACh interaction may be involved in serotonergic treatment of depression.     

Reversal of dexfenfluramine-induced anorexia and c-Fos/c-Jun expression by lesion in the lateral parabrachial nucleus

The external subdivision of the lateral parabrachial nucleus (LPBE) shows strong Fos-like immunoreactivity (FLI) following anorectic doses of the indirect serotonin agonist dexfenfluramine (DFEN). In an effort to determine the contribution of the LPBE to DFEN-induced anorexia, bilateral ibotenate lesions were made in the LPBE, and the effects of the lesion on DFEN-induced anorexia and FLI as well as c-June-like immunoreactivity (JLI) were examined. It was found that LPBE lesion significantly attenuated DFEN anorexia: in a 1-h food intake test following 24-h food deprivation, DFEN (2 mg/kg) suppressed food intake by 60% in intact rats but only 34% in rats with LPBE lesions. In addition to this behavioral change, LPBE lesion completely abolished DFEN-induced FLI and JLI in the lateral subdivision of the central nucleus of the amygdala (CeL) and laterodorsal subdivision of the bed nucleus of stria terminalis (BSTLD), both of which showed strong FLI and JLI in intact rats. DFEN-induced FLI and JLI in other brain regions were not affected by LPBE lesion, including the ventromedial and lateral hypothalamus, caudate-putamen, and the nucleus of the solitary tract (NST). The parallel loss of DFEN-induced anorexia and FLI/JLI following LPBE lesion raises the novel possibility that LPBE-CeL/BSTLD pathway may be involved in DFEN anorexia.     

Tryptophan hydroxylase in hippocampus and midbrain following unilateral injection of 5,7-dihydroxytryptamine

We have previously shown using anatomical methods that partial denervation of the rat hippocampus by removal of serotonergic (5-HT) fibers in the cingulum bundle induces sprouting of intact 5-HT fibers reaching the hippocampus in the fornix-fimbria. The biochemical properties of collateral sprouting fibers have remained largely uncharacterized. Thus, the rate-limiting enzyme in 5-HT synthesis, tryptophan hydroxylase (TPOH), was studied to determine whether new sprouts possess the ability to synthesize 5-HT. Unilateral stereotaxic injections of 5 micrograms 5,7-dihydroxytryptamine were made into the cingulum bundle of adult rats in order to produce partial and selective deafferentation of the hippocampus. Following injection, enzyme activity in the hippocampus declined gradually and bilaterally, reaching minimal levels by 7 days post-lesion. This decrease in enzyme activity was paralleled by a decrease in the Vmax and an increase in the Km of TPOH for tryptophan. Enzyme activity began to increase after 14 days post-lesion, reaching maximal levels by 60 days, but never returning to pre-lesion levels in the ipsilateral side. In midbrain, site of neuronal cell bodies of hippocampal 5-HT projections, enzyme activity gradually increased, reaching a maximum by 28 days after the lesion. These results indicate that 5-HT fibers remaining in the hippocampus following partial denervation are able to compensate biochemically for those removed by cingulum bundle lesions. Biochemical compensation may depend on increased synthesis of TPOH molecules in midbrain cell bodies with subsequent transport into sprouts of intact fibers in the hippocampus.     

Short-term effects of fluoxetine and trifluoromethylphenylpiperazine on electroencephalographic sleep in the rat

Fluoxetine and trifluoromethylphenylpiperazine (TFMPP) were studied for their short-term effects on electroencephalographic sleep in male rats. Following single injection, each drug produced a sizeable, dose-related suppression of rapid-eye-movement (REM) sleep that persisted for 4-5 h (fluoxetine, 0.625-5 mg/kg; TFMPP, 0.10-1.25 mg/kg). TFMPP also consistently increased non-REM (NREM) sleep during the second hour after drug injection, though this effect was not dose-related (it was seen at all doses tested). Fluoxetine produced small effects on NREM sleep that varied non-systematically with dose and time after drug injection. TFMPP, but not fluoxetine, also increased at all doses the number of delta waves per minute of NREM sleep in the second hour. A structural analog of TFMPP that is inactive at serotonin (5-HT) receptors [4-(m-trifluoromethylphenyl)piperadine; LY97117] was also tested, and found to be devoid of effects on NREM and REM sleep. Both fluoxetine (a 5-HT reuptake blocker) and TFMPP (a 5-HT agonist) enhance transmission across 5-HT synapses, though by different mechanisms. Because they have the common effect of suppressing REM sleep, and in a dose-related manner, the data support the notion that 5-HT neurons in the brain, when active, can suppress REM sleep.     

Chronic administration of the 5-HT1A receptor agonist 8-OH-DPAT differentially desensitizes 5-HT1A autoreceptors of the dorsal and median raphe nuclei

The present study investigated alterations of the regulation of serotonin (5-hydroxytryptamine; 5-HT) release by 5-HT_1A autoreceptors following single and repeated treatment with the 5-HT_1A receptor agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT). Rats were pretreated with 8-OH-DPAT (1.0 mg/kg, s.c.) for 1, 7, or 14 days. The ability of an acute challenge administration of 8-OH-DPAT (1.0 mg/kg, i.p.) to decrease 5-HT release in the ventral striatum and the ventral hippocampus of rats maintained under chloral hydrate anesthesia was examined 24 h after the last pretreatment injection using in vivo microdialysis. The decrease of 5-HT release in the striatum produced by the challenge dose of the 5-HT_1A receptor agonist was diminished following 7 and 14 days of pretreatment, but not after 1 day of pretreatment, with 8-OH-DPAT. In contrast, decreases of 5-HT release in the hippocampus by the 8-OH-DPAT challenge were not altered after 1 or 7 days of pretreatment, and only a trend for attenuation appeared after pretreatment for 14 days. The results of the present study indicate that desensitization of 5-HT_1A autoreceptors regulating 5-HT release in different brain regions by repeated treatment with 8-OH-DPAT occurs at different rates. Synapse 25:107–116, 1997. © 1997 Wiley-Liss, Inc.     

Effects of 5-HT and 8-OH-DPAT on forebrain monoamine synthesis after local application into the median and dorsal raphe nuclei of the rat

Summary 5-HT (10 and 40 g) and 8-OH-DPAT (1 and 5 g) were locally applied into the dorsal or median raphe nuclei in awake, unrestrained, rats. All animals were also treated with the 5-HTP and DOPA decarboxylase inhibitor NSD-1015, 100mg kg^–1 SC, 30 min before decapitation. 5-HT or 8-OH-DPAT were administered 5 min before NSD-1015. The regional brain in vivo rate of tyrosine and tryptophan hydroxylase activity was estimated by measuring the accumulation of DOPA and 5-HTP. The following brain regions were sampled: neocortex, hippocampus, dorso-lateral neostriatum, ventro-medial neostriatum, nucleus accumbens, olfactory tubercle, globus pallidus, septum and the amygdala.Compared to normal controls, there were small and inconsistent effects on forebrain 5-HTP accumulation by saline injections into the dorsal or the median raphe (an increase in 3 out of 36 experiments), whereas strong effects by the injection procedure were noted on forebrain DOPA accumulation (an increase in 15 out of 36 experiments).Injections of 5-HT (same effect by 10 or 40 g) into the dorsal raphe, produced a decrease in 5-HTP accumulation in all forebrain areas except for the hippocampus and the septum, whereas no effects were seen in any area after median raphe injections. In contrast, 8-OH-DPAT preferentially produced a decrease in forebrain 5-HTP accumulation after median raphe injections and less, but statistically significant effects by dorsal raphe injections. The 8-OH-DPAT injection into the median raphe primarily affected limbic forebrain areas (hippocampus, nucleus accumbens, ventro-medial neostriatum, amygdala and the septum).This dissociation of the effects of 5-HT and 8-OH-DPAT on forebrain 5-HT synthesis after local application into the dorsal or the median raphe strongly supports the contention of heterogeniety in the brain 5-HT receptor population in terms of receptor subtypes and/or receptor regulation.     

Circadian rhythm in the response to intracerebroventricular administration of 8-OH-DPAT

In a recent study, we observed circadian rhythms in the response to subcutaneous (s.c.) administration of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a serotonin (5-HT)_1A receptor agonist, in rats. Although these results suggested a circadian rhythm in the function of the central 5-HT_1A receptor, it was not possible to completely exclude drug disposition as the generating factor. The present study investigated circadian rhythms in the behavioral responses to intracerebroventricular administration of 8-OH-DPAT in rats. The results indicated the existence of circadian rhythms of a similar pattern to those observed in the forepaw treading and head weaving responses to s.c. administration of the drug. The flat body posture response observed following s.c. drug administration was not evident in the present study. These observations suggest that the central postsynaptic 5-HT_1A receptor exhibits a circadian rhythm in its function.     

Anxiogenesis in adult rats treated chronically with cocaine during adolescence: effects of extended abstinence and 8-OH-DPAT treatment

Our laboratory has recently observed the development of an anxiogenic response after a short abstinence period (10 days) in young adult rats treated repeatedly with cocaine during adolescence. The present study was conducted to determine if this effect persists into adulthood following extended durations of abstinence and whether it could be modulated with the 5-HT_1A agonist (±)-8-hydroxy-2-(dipropylamino)tetralin hydrobromide (8-OH-DPAT). Accordingly, 30-day-old rats were injected with either 10 mg/kg cocaine or saline for 8 consecutive days. Approximately 8 weeks after the final injection, anxiety levels in subjects were assessed with an elevated zero maze with a second assessment performed 4 weeks thereafter. Shortly prior to each test session, half the subjects in each of the two adolescent drug conditions received injections of 300 μg/kg 8-OH-DPAT while the other half received injections of the vehicle. Based on total time spent in the open areas of the maze, the results obtained at the first abstinent duration indicated that adolescent cocaine treatment did not induce an anxiogenic response. Assessment of maze behavior at the second abstinent duration was aided with a digital tracking and computerized scoring system (LimeLight, Actimetrics). Similar to the results obtained at the first abstinent duration, the amount of time spent in the open areas of the maze was unrelated to prior cocaine treatment. However, cocaine-treated rats did show evidence of an anxiogenic response at this abstinent duration based on more frequent entries into and out of the open and enclosed areas of the maze, more frequent and longer durations of exploratory bouts beyond the perimeter of the maze, excessive number of cautious protrusions into the open areas, and faster running velocities through the open areas. These results were not artifacts of generalized motor activation in that comparable total distance traveled scores were noted for all subjects. Treatment with 8-OH-DPAT shortly prior to elevated zero maze testing normalized these behaviors induced by adolescent cocaine. It is concluded that the anxiogenic response produced by cocaine exposure during adolescence persists 12 weeks beyond cessation of drug treatment and that this effect is reversible with 8-OH-DPAT. In addition, the results underscore the importance of taking multiple measures when assessing anxiety in experimental animals. Implications for the neurobiology of drug abuse and the role of serotonin are discussed.     

Progesterone attenuates the effect of the 5-HT1A receptor agonist, 8-OH-DPAT,and of mild restraint on lordosis behavior

Ovariectomized, hormone-primed rats were used to test the hypothesis that progesterone treatment attenuated the effects of the 5-HT(1A) receptor agonist, (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), on female rat lordosis behavior. Based upon prior evidence that prepriming with estradiol benzoate (EB) reduced the ability of 8-OH-DPAT to inhibit lordosis behavior, rats were preprimed with 10 microg EB 7 days before a second priming with 10 microg EB followed 48 h later with 500 microg progesterone or vehicle. Independent of the presence of progesterone, prepriming with EB attenuated the lordosis-inhibiting effects of systemic treatment with 8-OH-DPAT. However, progesterone also reduced the effects of 8-OH-DPAT and this effect was also seen in females primed only once with EB. In contrast, progesterone was relatively ineffective in attenuating the effects of bilateral infusion with 8-OH-DPAT into the ventromedial nucleus of the hypothalamus (VMN). The failure of progesterone to substantially reduce the effects of VMN infusion with 8-OH-DPAT contrasts with prior studies in which estrogen's protective action against the drug did include the VMN. Thus, while both estrogen and progesterone reduce the lordosis-inhibiting effect of 8-OH-DPAT, the mechanisms responsible for the effects of the two gonadal hormones may be different. Priming with progesterone also prevented the effects of 5 min of restraint. When rats were hormonally primed with EB and oil, rats showed a transient, but significant, decline in lordosis behavior 5 and 10 min after restraint. Rats primed with EB and progesterone were unaffected by the restraint. These results are discussed in terms of their implications for the role of progesterone in altering the 5-HT(1A) receptor modulation of lordosis behavior.     

Sleep effects following intrathecal administration of the 5-HT1A agonist 8-OH-DPAT and the NMDA antagonist AP-5 in rats

The modulating effect of an intrathecally (i.t.) administered 5-HT_1A agonist and an NMDA antagonist on sleep, waking and EEG power spectra was investigated in rats. The 5-HT_1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) (38 nmol) increased total slow wave sleep (TSWS) and decreased waking over the 8 h recording period. The TSWS increase was mostly due to an increase in SWS1. Sleep latency to SWS1 was also reduced. The NMDA antagonist dl-2-amino 5-phosphonovaleric acid (AP-5) (31.5 nmol) reduced waking. SWS1 was increased, but TSWS was not changed. An increase in REM sleep was seen during the last part of the recording. Combined treatment with 8-OH-DPAT and AP-5 reduced waking and increasd TSWSS. No change in REM sleep was seen. There were no systematic changes in either waking, TSWS or REM fronto-frontal or fronto-parietal EEG power spectrum after any of the treatments. The results suggest that in the spinal cord stimulation of 5-HT_1A receptors have a dampening effect on transmission of sensory information, leading to deactivation and thereby increased possibilities for sleep induction. Blockade of the NMDA receptors may also lead to a small dampening of sensory transmission with similar consequences.     

Baseline and 8-OH-DPAT-induced release of acetylcholine in the hippocampus of aged rats with different levels of cognitive dysfunction

During aging, neurotransmission systems such as the cholinergic and serotonergic ones are altered. Using rats aged 3 or 24-26 months, this study investigated whether the well-described 8-OH-DPAT-induced increase of hippocampal acetylcholine release was altered in aged rats and whether it may vary according to the magnitude of age-related cognitive deficits. Long-Evans female rats aged 24-26 months were classified as good or bad performers on the basis of their reference-memory performance in a Morris water-maze task. Subsequently, the efficiency of 5-HT(1A) receptor agonist 8-OH-DPAT (0.5 mg/kg, s.c.) in triggering hippocampal acetylcholine release was evaluated by in vivo microdialysis and high performance liquid chromatography analysis. Besides a reduced baseline release in aged rats and a correlation between the baseline release and probe-trial performance in all rats, the results demonstrated that 8-OH-DPAT produced a significant increase of hippocampal acetylcholine release (peak value) in all rats, whether aged or young. While significant in bad performers (+56%), this increase did not reach significance in good performers (+32%). The results suggest that (i) some aspects of cognitive alterations related to aging might be linked to the baseline release of acetylcholine in the hippocampus, and (ii) the cholinergic innervation of the hippocampus of aged rats responds almost normally to systemic activation of 5-HT(1A) receptors, and (iii) differential alterations of cholinergic/serotonergic interactions assessed by determination of the 8-OH-DPAT-induced release of acetylcholine in the hippocampus could not be linked with clarity to the cognitive status of aged rats.     

Effects of 21 days treatment with fluoxetine on stimulated endogenous 5-hydroxytryptamine overflow in the rat dorsal raphe and suprachiasmatic nucleus studies using fast cyclic voltammetry in vitro

Changes in the extracellular concentration of 5-HT evoked by electrical stimulation of brain slices containing either dorsal raphe nucleus (DRN) or suprachiasmatic nucleus (SCN) from rats treated for 21 days with fluoxetine (5 mg/kg; i.p.) or water were monitored using fast cyclic voltammetry (FCV). Stimulated 5-HT overflow was enhanced significantly in both brain regions after 21 days treatment with fluoxetine but there was no change in the half time for re-uptake (t1/2). Concentration response curves for inhibition of electrically stimulated 5-HT overflow by 8-OH-DPAT (5-HT_1a receptor agonist) or RU24969 (5-HT_1b receptor agonist) in the DRN or SCN respectively were obtained in slices prepared from both groups of animals. There was a significant shift to the right in the dose-response curve for RU24969 in the SCN in fluoxetine treated animals but a shift to the left for the dose-response curve for 8-OH-DPAT in the DRN. These data suggest that down regulation of the 5-HT_1b autoreceptors occurs in an axon terminal region (SCN) but that there is a sensitisation of 5-HT_1a autoreceptor mechanisms controlling 5-HT overflow in the DRN.     

Release of serotonin induced by 3,4-methylenedioxymethamphetamine (MDMA) and other substituted amphetamines in cultured fetal raphe neurons: further evidence for calcium-independent mechanisms of release

The substituted amphetamines 3,4-methylenedioxyamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), p-chloroamphetamine (PCA) and fenfluramine (FEN) all exert their effects by releasing serotonin (5-HT) from presynaptic nerve terminals. In the current study, we examined the ability of these agents to induce the release of 5-HT in cultured fetal raphe neurons. The data indicate that the rank order of release potencies for these agents was (±)PCA > (+)MDMA = (+)MDA = (±)FEN. Studies examining the role of calcium in 5-HT release demonstrate that preventing calcium influx with L- and N-type calcium channel blockers inhibits potassium-stimulated release of [³H]5-HT but has no effect on release induced by the substituted amphetamines. Furthermore, omitting calcium from the extracellular media or depleting the vesicular pool of neurotransmitter with continual potassium stimulation did not affect the release of [³H]5-HT induced by these compounds. Administration of fluoxetine prior to the substituted amphetamines significantly attenuated the releasing effects of these agents, while producing no effect on potassium-stimulated release. These results are consistent with the notion that the amphetamines induce release of cytoplasmic 5-HT via the plasma membrane transporter.