8-OH-DPAT influences extracellular levels of serotonin and dopamine in the medial preoptic area of male rats

Serotonin (5-HT) is generally inhibitory to male rat sexual behavior. However, the 5-HT_1A agonist 8-hydroxy-di-propylaminotetralin (8-OH-DPAT), injected either systemically or into the medial preoptic area (MPOA), facilitates ejaculation. Three experiments were conducted to test the effects of 8-OH-DPAT on 5-HT and dopamine (DA) neurotransmission in the MPOA, a very important site for the control of male sexual behavior. In Experiment 1, systemically injected 8-OH-DPAT (0.4 mg/kg) decreased extracellular 5-HT levels in the MPOA as measured by in vivo microdialysis. In Experiment 2, 8-OH-DPAT (500 μM) administered directly into the MPOA via reverse dialysis increased extracellular levels of both DA and 5-HT; pretreatment with the selective 5-HT_1A antagonist 4-iodo-N-[2-[4-(methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinyl-benzamide hydrochloride (p-MPPI) failed to prevent 8-OH-DPAT's stimulatory effects on DA and 5-HT levels in the MPOA. In Experiment 3, 8-OH-DPAT (8 μg) co-injected with 5,7-dihydroxytryptamine (5,7-DHT; 6 μg) prevented neurotoxic depletion of 5-HT in the site of injection (MPOA). Because systemic and MPOA injections of 8-OH-DPAT resulted in opposite effects on extracellular 5-HT in the MPOA, yet both can facilitate ejaculation, these data suggest that moderate changes in 5-HT in the MPOA may have relatively little influence on male copulatory behavior. Instead, the facilitative effects of 8-OH-DPAT in the MPOA on male copulatory behavior may result, at least in part, from stimulatory effects of 8-OH-DPAT on DA transmission. Facilitative effects of systemic injections of 8-OH-DPAT may result from decreased 5-HT release in several sites.     

Partial antagonism of 8-OH-DPAT'S effects on male rat sexual behavior with a D2, but not a 5-HT1A, antagonist

The serotonin agonist 8-hydroxy-di-propylaminotetralin (8-OH-DPAT), injected systemically or directly into the medial preoptic area (MPOA), reduces the ejaculatory threshold in male rats. While 8-OH-DPAT has been characterized as an agonist at the 5-HT_1A receptor, it also acts at other receptor sites including the dopamine D₂ receptor. The current experiments investigated whether 8-OH-DPAT injected into the MPOA facilitates male sexual behavior through stimulation of the 5-HT_1A receptor or the dopamine D₂ receptor. Experiment 1 co-administered 8-OH-DPAT (6 μg) with either the 5-HT_1A antagonist 4-iodo-N-[2-[4-(methoxyphenyl)-1-piperazinyl]ethyl]-N-2-pyridinyl-benzamide hydrochloride (MPPI) (10 μg) or the D₂ antagonist raclopride (10 μg). Raclopride blocked 8-OH-DPAT's facilitative effects on ejaculation frequency and latency, while the 5-HT_1A antagonist was ineffective. In Experiment 2, 8-OH-DPAT (500 μM), retrodialyzed into the MPOA through a microdialysis probe, enhanced male copulatory behavior similarly to the microinjection, increasing ejaculation frequency and decreasing ejaculation latency, postejaculatory interval and mount frequency. Retrodialyzing 8-OH-DPAT through a microdialysis probe in the MPOA had been previously shown to increase extracellular levels of dopamine and serotonin. The data from the present studies suggest that the effects of 8-OH-DPAT in the MPOA on male rat copulatory behavior may be mediated, at least in part, either directly through 8-OH-DPAT's activity at D₂ receptors or indirectly through 8-OH-DPAT's ability to increase extracellular dopamine.     

Long-term fluoxetine, but not desipramine, inhibits the ACTH and oxytocin responses to the 5-HT1A agonist, 8-OH-DPAT, in male rats

The present studies determined whether serotonin 5-HT_1A receptor-mediated function is modified by chronic exposure to antidepressants. Hormone responses to the 5-HT_1A agonist, 8-OH-DPAT, were evaluated after long-term exposure to two antidepressants, the 5-HT uptake blocker, fluoxetine, and the norepinephrine uptake blocker, desipramine (DMI). In addition, the density and affinity of 5-HT_1A receptors in the hypothalamus and cerebral cortex were measured. Male rats received fluoxetine (10 mg/kg i.p.), DMI (5 mg/kg i.p) or saline injections once daily for 21 days. 8-OH-DPAT (0–500 μg/kg s.c.) was administered 18 h after the final antidepressant injection and 15 min before sacrifice. 8-OH-DPAT significantly increased plasma ACTH, corticosterone, oxytocin and prolactin, but not renin or vasopressin concentrations. Chronic injections of fluoxetine inhibited the ACTH, corticosterone and oxytocin responses to 8-OH-DPAT. suggesting reduced 5-HT_1A receptor function. In contrast, chronic DMI did not alter the hormone responses to 8-OH-DPAT. The density and affinity of 5-HT_1A receptors in the frontal cortex or hypothalamus were not altered by either fluoxetine or DMI. To verify that the observed effects require prolonged exposure to fluoxetine, rats received a single injection of fluoxetin (10 mg/kg, i.p.), 3 h before 8-OH-DPAT (0–500 μg/kg s.c.). Acute fluoxetine did not reduce any of the hormone responses to 8-OH-DPAT. In conclusion, the results suggest that chronic, but not acute, exposure to fluoxetine decreases 5-HT_1A receptor function. This effects is not seen in rats chronically exposed to DMI. The mechanism of the effects of chronic fluoxetine on 5-HT_1A receptor-mediated hormone responses is not likely due to reduced density of 5-HT_1A receptors, because no changes were observed in [³H]8-OH-DPAT binding in hypothalamus and cortex, but could be due to changes in signal transduction.     

Differential serotonergic inhibition of in vitro striatal [3H]acetylcholine release in prenatally cocaine-exposed male and female rats

Previous research indicates that prenatal cocaine (pCOC)-exposure results in greater 5-HT₃ agonist-induced inhibition of electrically evoked [³H]acetylcholine (ACh) overflow in rat striatal slices. The present study examines the effects of fluoxetine (FLU)-induced and exogenous serotonin (5-HT) on electrically evoked ACh release from striatal slices prepared from adult male and female (in periods of diestrus or proestrus) rats exposed to saline or cocaine in utero. Additionally, we assessed the impact of monoaminergic receptor stimulation on evoked ACh release by superfusion with selective 5-HT₂, 5-HT₃ and D₂ receptor antagonists in the presence of FLU-induced and exogenous 5-HT and measuring the capacity of these drugs to reverse inhibitory effects of 5-HT. Given our previous findings of accentuated inhibition of ACh release by 5-HT₃ agonism in striata of pCOC-exposed adult rats, we hypothesized that superfusion of endogenous and exogenous 5-HT would lead to greater suppression of evoked ACh release in this group of animals. Our results indicated that ACh release from slices of all prenatal saline (pSAL) rats was inhibited comparably by FLU (10 μM)-elicited increases in endogenous 5-HT or by increases elicited with application of exogenous 5-HT (5 μM). Robust FLU-mediated inhibition of ACh release was evident in slices from pCOC male and pCOC diestrus female rats vs. their respective PSAL control groups. Superfusion of striatal slices with 5-HT (5 μM) produced a pattern of ACh inhibition similar to that produced by FLU; however, the magnitude of ACh inhibition was consistently greater than that observed with FLU. Inhibition of ACh overflow by FLU was blocked by co-superfusion with ketanserin, a 5-HT₂ receptor antagonist, ICS-205,930, a 5-HT₃ receptor antagonist or sulpiride, a D₂ receptor antagonist. Conversely, serotonergic inhibition of ACh overflow was only blocked by a high concentration of ICS-205,930 (5 μM) and was completely reversed by sulpiride (1 μM). Collectively, these findings demonstrate serotonergic modulation of cholinergic neurons varying as a function of prenatal treatment, sex and, for females, phase of estrous. Inhibition of ACh release by 5-HT appears to be mediated by a complex relationship between 5-HT₂, 5-HT₃ and D₂ receptor regulation, as the blockade of any of these receptors reversed the inhibitory effects of FLU on ACh release. Conversely, in the case of exogenous 5-HT-induced inhibition, only blockade of D₂ receptors and high concentrations of the 5-HT₃ receptor antagonists were capable of reversing monoaminergic inhibition. These data support the hypothesis that the enhanced serotonergic modulation of ACh neurons in pCOC-exposed animals is largely mediated by dopamine (DA) and reflect a major biochemical persistence of neurodevelopmental adaptations elicited by early cocaine exposure.     

Pharmacological characterization of grooming induced by a selective NK-1 tachykinin receptor agonist

Bilateral intranigral administration of the selective NK-1 tachykinin receptor agonist [AcArg⁶, Sar⁹, Met(O₂)¹¹]SP_6–11 (0–11 nmol total bilateral dose) selectively induced grooming in rats. This response was blocked by concurrent intranigral administration of the NK-1 tachykinin receptor antagonist RP 67580 (2 nmol), but not by NK-2 (L-659, 877) or NK-3 ([Trp⁷, β-Ala⁸]NKA_4–10) antagonists. Pretreatment with systemic opioid (naloxone 1.5 mg/kg) and D₁ dopamine (SCH 23390 100 μg/kg) receptor antagonists also attenuated tachykinin-induced grooming, which was unaffected by D₂ dopamine (sulpiride 30 mg/kg) or 5-HT_2A+C (ritanserin 2 mg/kg) antagonists. Grooming induced by intranigral [AcArg⁶, Sar⁹, Met(O₂)¹¹]SP_6–11 was also attenuated by bilateral 6-hydroxydopamine lesions of te substantia nigra. These findings indicate that grooming induced by intranigral tachykinins reflects activation of NK-1 receptors and is dependent upon endogenous dopamine and consequent selective stimulation of D₁ dopamine receptors.     

Sleep/waking effects of a selective 5-HT1A receptor agonist given systemically as well as perfused in the dorsal raphe nucleus in rats

Sleep/waking stages and behavior were studied following the selective 5-HT_1A agonist 8-OH-DPAT given subcutaneously (s.c.) (0.010–0.375 mg/kg) as well as perfused continuously (10 μM) for 6 h into the dorsal raphe nucleus (DRN) using microdialysis. Given systemically, 8-OH-DPAT at 0.375 mg/kg s.c. induced 5-HT behavioral syndrome, increased waking to 149% and reduced slow wave sleep (SWS) to 86%, transition to 76% and rapid eye movement (REM) sleep to 73%. The effect on deep SWS (SWS-2) was biphasic, with an increase after 2 h. 8-OH-DPAT at 0.010 mg/kg did not have any vigilance effects. 8-OH-DPAT perfusion in DRN produced a fourfold increase in REM sleep compared to perfusion of artificial cerebrospinal fluid. This is consistent with the hypothesis that reduced 5-HT neurotransmission following 5-HT_1A autoreceptor stimulation will disinhibit cholinergic REM-promoting mesopontine neurons and thereby lead to a REM sleep increase. The other sleep/waking stages were not significantly affected by 8-OH-DPAT perfusion in DRN.     

Autoradiographic analyses of 5-HT1A and 5-HT2A receptors after social isolation in mice

Social isolation of rodents is used to model human psychopathological processes. In the present study, the effects of intermediate and long term isolation housing on postsynaptic 5-HT_1A and 5-HT_2A receptors were analyzed in male mice housed in groups or isolation for 4 and 12 weeks. [³H]8-OH-DPAT and [³H]ketanserin were used to label 5-HT_1A and 5-HT_2A receptors. Four representative sagittal sections (planes 1–4) were scored by in vitro autoradiography. Whereas after 4 weeks of housing both receptor densities were lowered significantly in isolated mice, after 12 weeks of housing only marginal isolation effects were seen. Intermediate isolation reduced 5-HT_1A receptors especially in the lateral frontal, parietal and entorhinal cortex (−63%), in the lateral CA1–3 and dentate gyrus region of the hippocampus (−68%), in the basolateral, basomedial, central and medial amygdaloid nuclei (between −38 and −66%), and in the hypothalamus (−28%). 5-HT_2A receptors were strongly reduced in the frontal cortex (between −47 and −74%), in the hippocampus (between −47 and −95%), in the striatum (between −66 and −76%), and in the accumbens nucleus (between −59 and −73%) in comparison to group housed control mice. After 12 weeks of isolation in the hippocampus continuously decreased 5-HT_1A receptor densities were demonstrated (between −24 and −61%). But increased 5-HT_2A receptor densities were seen in the lateral striatum (+86%) compared to control mice. Age-dependent effects were also found. After 12 weeks of group housing the 5-HT_1A and 5-HT_2A receptor densities were decreased (between −28 and −54%) in all analyzed brain regions in comparison to 4 weeks of group housing. Isolated animals showed diminished 5-HT_1A receptor densities in the cortex (−14%) and hippocampus (−15%), but increased 5-HT_1A receptor densities in the amygdala (+33%) after 12 weeks. The 5-HT_2A receptor densities were increased in all analyzed regions (between +31 and +96%) after 12 weeks of isolation compared to 4 weeks. To explain these dynamic, time-dependent pattern of isolation-induced changes different regulation processes are supposed regarding 5-HT_1A and 5-HT_2A receptors. Besides metabolism-related adaptation processes also neurotransmitter and hormonal (e.g., glucocorticoid) interactions especially in limbic regions have to be considered.     

5-HT-Dependent myoclonus in guinea pigs: Mediation through 5-HT1A-5-HT2 receptor interaction

Investigations utilizing agonists for 5-HT receptor subtypes have been conducted to determine which 5-HT receptor subtype(s) subserve myoclonus in the guinea pig. Administration of a nonselective 5-HT agonist such as 5-MeODMT (5-HT_1A/5-HT₂ agonist) induces a dose-dependent behavior characterized by head jerking at low doses (1–2 mg/kg, SC) and full-blown myoclonus (continuous rhythmic whole-body jerking) at higher doses (2.5–5 mg/kg, SC). In contrast, the selective 5-HT_1A receptor agonist 8-OH-DPAT and the selective 5-HT₂ receptor agonist DOI do not induce myoclonus, and elicit only limited head jerking across an otherwise behaviorally active range of doses (1–5 mg/kg, SC). Importantly, the coadministration of both 8-OH-DPAT and DOI results in the emergence of dose-dependent myoclonic behavior. These data suggest that coactivation of 5-HT,A and 5-HT2 receptors may be required for the induction of myoclonus in the guinea pig.     

Further evidence that extracellular serotonin in the rostral ventrolateral medulla modulates 5-HT(1A) receptor-mediated attenuation of exercise pressor reflex

We determined changes in extracellular levels of glutamate, serotonin (5-HT), norepinephrine (NE), and dopamine (DA) within rostral ventrolateral medulla (RVLM) during 5-HT_1A-receptor stimulation-mediated inhibition of cardiovascular responses to static muscle contraction using anesthetized rats. In ten rats, muscle contraction significantly increased (P<0.01) mean arterial pressure (MAP) by 29±4 mmHg, heart rate (HR) by 25±3 bpm, and glutamate levels by 4.5±0.8 ng/5 μl. Microdialysis of a 5-HT_1A receptor agonist, 8-OH-DPAT (10 mM), into the RVLM for 30 min attenuated cardiovascular responses to muscle contraction and had no effect on glutamate levels. A subsequent administration of 10 mM WAY100635, a 5-HT_1A antagonist, into the RVLM antagonized the attenuating effects of 8-OH-DPAT. In another ten rats, muscle contraction significantly increased (P<0.01) MAP and HR by 20±2 mmHg and 25±8 bpm, respectively. In addition, levels of 5-HT, NE, and DA in the RVLM significantly increased (P<0.01) by 3.6±0.3, 3.2±0.3, and 3.3±0.4 pg/10 μl, respectively. Administration of 8-OH-DPAT (10 mM) into the RVLM for 30 min attenuated cardiovascular responses to muscle contraction and had no effects on NE and DA levels. However, the drug significantly attenuated 5-HT levels following a muscle contraction. Microdialysis of 10 mM WAY100635 into the RVLM reversed both cardiovascular and 5-HT changes. These results suggest that stimulation of 5-HT_1A-receptors within the RVLM attenuates cardiovascular responses to static exercise via a reduction of extracellular 5-HT concentration and most likely not through changes in glutamate, NE or DA levels.     

Relationship of increased food intake and plasma ACTH levels to 5-HT1A receptor activation in rats

Various putative agonists of the 5-HT_1A receptor subtype induce feeding in rats, probably by activating raphé somatodendritic 5-HT autoreceptors. These drugs also produce a marked increase in plasma concentrations of corticotropin (ACTH). In the present experiment we attempted to localize the site of action of 5-HT_1A agonists on the secretion of ACTH and examined the relationship between 5-HT_1A agonist-induced feeding and ACTH secretion. Rats were injected with either the high affinity 5-HT_1A agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) (0.016-1.0 mg/kg, s.c.) or the novel anxiolytics buspirone, gepirone or ipsapirone (2.0–16.0 mg/k/g, s.c.), and either had their food intake measured 2 hr post injection or were sacrificed 30–40 min post injection for measurement of plasma ACTH. Plasma ACTH also was measured in rats pretreated with the serotonin synthesis inhibitor, para-chlorophenylalanine (PCPA) for three days (150 mg/kg, i.p. per day) and subsequently injected with 8-OH-DPAT (0.3 mg/kg, s.c.).

As previously reported, the 5-HT_1A agonists increased both food agonists increased both food intake and plasma ACTH concentrations. After 8-OH-DPAT, ipsapirone and gepirone the amount of food consumed was positively correlated with the concentration of plasma ACTH. No such correlation was evident following buspirone. PCPA pretreatment resulted in near total depletion of brain 5-HT content but had no effect on the ACTH rise induced by 8-OH-DPAT. Therefore, in contrast to the presynaptic site previously proposed for 5-HT_1A agonist-induced feeding, the present results suggest a agonist-induced feeding, the present results suggest a postsynaptic location for the 5-HT_1A receptor mediating ACTH release.     

Suppression of the micturition reflex in urethane-anesthetized rats by intracerebroventricular injection of WAY100635, a 5-HT(1A) receptor antagonist

The influence of supraspinal 5-HT_1A receptors on reflex bladder activity was evaluated in anesthetized rats by studying the effects of intracerebroventricular (i.c.v.) administration of WAY100635 (1–100 μg), a selective 5-HT_1A receptor antagonist. The drug dose-dependently decreased the frequency and/or amplitude of isovolumetric reflex bladder contractions. Low doses (1–10 μg) increased the interval between contractions but only slightly reduced the amplitude of the contractions. However, 100 μg of WAY100635 elicited an initial complete block of bladder reflexes followed by a recovery period lasting 10–15 min during which the frequency of reflex contractions was normal but the amplitude was markedly suppressed by 70–80%. Mesulergine (0.1 mg/kg, i.v.), a 5-HT_2C antagonist, which transiently eliminated bladder activity in some rats (five of 11), blocked the inhibitory effect of WAY100635 (10 or 100 μg, i.c.v.) in only two of six rats. Our data coupled with the results of previous studies suggest that spinal and supraspinal 5-HT_1A receptors are involved in multiple inhibitory mechanisms controlling the spinobulbospinal micturition reflex pathway. The regulation of the frequency of bladder reflexes is presumably mediated by a suppression of afferent input to the micturition switching circuitry in the pons, whereas the regulation of bladder contraction amplitude may be related to an inhibition of the output from the pons to the parasympathetic nucleus in the spinal cord.     

Melatonin reversal of DOI-induced hypophagia in rats; possible mechanism by suppressing 5-HT(2A) receptor-mediated activation of HPA axis

Serotonin type 2A (5-HT_2A) receptor-mediated neurotransmitter is known to activate hypothalamic–pituitary–adrenal (HPA) axis, regulate sleep–awake cycle, induce anorexia and hyperthermia. Interaction between melatonin and 5-HT_2A receptors in the regulation of the sleep–awake cycle and head-twitch response in rat have been reported. Previous studies have shown that melatonin has suppressant effect on HPA axis activation, decreases core body temperature and induces hyperphagia in animals. However, melatonin interaction with 5-HT_2A receptors in mediation of these actions is not yet reported. We have studied the acute effect of melatonin and its antagonist, luzindole on centrally administered (±)-1-(2,5-dimethoxy-4-iodophenyl) 2-amino propane (DOI; a 5-HT_2A/2C agonist)-induced activation of HPA axis, hypophagia and hyperthermia in 24-h food-deprived rats. Like ritanserin [(1 mg/kg, i.p.) 5-HT_2A/2C antagonist], peripherally administered melatonin (1.5 and 3 mg/kg, i.p.) did not affect the food intake, rectal temperature or basal adrenal ascorbic acid level. However, pretreatment of rats with it significantly reversed DOI (10 μg, intraventricular)-induced anorexia and activation of HPA axis. But the hyperthermia induced by DOI was not sensitive to reversal by melatonin. Mel₁ receptor subtype antagonist luzindole (5 μg, intraventricular) did not modulate the DOI effect but antagonized the melatonin (3 mg/kg, i.p.) reversal of 5-HT_2A agonist response. The present data suggest that melatonin reversal of DOI-induced hypophagia could be due to suppression of 5-HT_2A mediated activation of HPA axis.     

Anxiolytic-like effects of 5-HT2 ligands on three mouse models of anxiety

The behavioural effects of 5-HT₂ receptor agonists, 5-HT_2A and 5-HT_2C receptor antagonists were investigated in the mouse four plates test (FPT), light/dark paradigm (L/D) and the elevated plus maze (EPM), in order to elucidate the role of the 5-HT₂ receptor subtypes in these models and to address the inconclusive results previously reported using rat psychopharmacological models. All compounds were administered intraperitoneally 30 min before each test. DOI, a preferential 5-HT_2A agonist (0.5–8 mg/kg) and BW 723C86, a 5-HT_2B agonist (8 and 16 mg/kg) provoked an anxiolytic-like response in the FPT. In the EPM, an anxiolytic-like effect was observed for DOI (0.5, 1 and 2 mg/kg), BW 723C86 (0.5, 4, 8 and 16 mg/kg), RO 60-0175 a 5-HT_2C agonist (4 mg/kg) and the non-selective 5-HT₂ receptor agonist mCPP (0.25 mg/kg.). Ketanserin, a 5-HT_2A/2C non-selective receptor antagonist (0.015 and 0.03 mg/kg), induced an anxiogenic-like effect in the L/D paradigm. The 5-HT_2C antagonists (RS 10-2221, SDZ SER082 and SB 206553) were without effect in all three tests. These behavioural results are indicative of an anxiolytic-like action of 5-HT₂ receptor agonists, an anxiogenic-like effect of 5-HT_2A receptor antagonism, whereas the blockade of 5-HT_2C receptors are without effect in the mouse models studied.     

Presynaptic D2 dopaminergic receptors mediate inhibition of excitatory synaptic transmission in rat neostriatum

The effect of dopamine (DA) on excitatory synaptic transmission was studied in rat neostriatal neurons using intracellular- and whole-cell voltage clamp-recording methods. Depolarizing excitatory postsynaptic potentials (EPSPs) were evoked by cortical stimulation. Superfusion of DA (0.01–10 μM) reversibly decreases EPSP in a concentration-dependent manner and with a estimated IC₅ of 0.3 μM. In addition, the inhibitory effect induced by DA at a low concentratiion (0.1 μM) was antagonized by sulpiride (1–10 nM), a selective D₂ dopaminergic receptor antagonist. However, D₁ dopaminergic receptor antagonist SKF-83566 (1–5 μM) did not affect the blocking effect by DA 0.1 μM. Based on these findings, we conclude that DA at a low concentration ( 0.1 μM) reduced the excitatory response of neostriatal neurons following cortical stimulation via the activation of D₂, but not D₁ dopaminergic receptors, located on the terminals of corticostriatal neurons.     

Inhibitory effects of tandospirone, a 5-HT1A agonist, on medial vestibular nucleus neurons responding to lateral roll tilt stimulation in rats

An electrophysiological study was performed using chloral hydrate-anesthetized rats to determine whether tandospirone, a 5-HT_1A agonist, affects neuronal activities of the medial vestibular nucleus (MVN), since serotonergic innervation and 5-HT_1A receptors are present in this nucleus. Tandospirone applied microiontophoretically at a current of 20–60 nA caused an inhibition of tilt-induced firing of -type neurons, which showed increased and decreased firing with lateral tilt ipsilateral and contralateral to the recording site, respectively, along with that of β-type neurons which exhibited the reverse responses to ipsilateral and contralateral tilt stimulation. The inhibition was antagonized during simultaneous, iontophoretic application of WAY-100635 (20–60 nA), a 5-HT_1A receptor antagonist, although WAY-100635 alone rarely affected spontaneous or tilt-induced firing in either type of neurons. These results suggest that tandospirone acts on a 5-HT_1A receptor to inhibit transmission of otolith information to - and β-type MVN neurons.     

Lack of potentiation of the anti-alcohol effects of fluoxetine by pindolol in alcohol-preferring caa rats

1. 1. Recent clinical evidence suggests that the nonselective 5-HT_1A receptor antagonist pindolol may enhance the antidepressive efficacy of selective serotonin reuptake inhibitors (SSRIs); an effect generally ascribed to a presumed blockade of somatodendritic 5-HT_1A receptors by pindolol.

2. 2. The present study investigated whether blockade of 5-HT_1A receptors similarly potentiates the previously reported anti-alcohol effects of the SSRI fluoxetine.

3. 3. Pindolol and the selective 5-HT_1A receptor antagonist WAY-100635 were tested alone and in combination with fluoxetine in cAA rats, a genetic model of alcoholism. However, as pindolol has also a high affinity to 5-HT_1B receptors, the effects of selective 5-HT_1B receptor agonists and antagonists were evaluated as well.

4. 4. Neither pindolol (3–30 mg/kg, IP), nor WAY-100635 (1–10 mg/kg, IP) affected alcohol intake when tested alone. In contrast, the 5-HTm receptor agonists CP-94,253 and TFMPP (both 1–10 mg/kg, IP), and antagonists metergoline (1–10 mg/kg, IP) and GR 127935 (3–30 mg/kg, IP) were found to reduce alcohol intake with different degrees of selectivity (that is, the extent to which reductions in ethanol intake could be separated from reductions in food- and/or total fluid intake) and specificity (that is, the degree to which effects on ethanol intake coincided with effects on ethanol preference).

5. 5. Because the behavioral profile of pindolol resembles that of WAY-100635, and not that of the 5-HT_1B receptors ligands, combination experiments with fluoxetine were only performed with the former two compounds. Neither pindolol (30 mg/kg), nor WAY-100635 (3 mg/kg) potentiated the anti-alcohol effects of fluoxetine (10 mg/kg, IP). Moreover, WAY-100635 tended to shift the anti-alcohol effect of fluoxetine towards a less selective and specific profile.

It is concluded that acute blockade of 5-HT_1A receptors does not potentiate the anti-alcohol effects of fluoxetine. In addition, it is suggested that different mechanisms underly the antidepressive and anti-alcohol effects of SSR.     

Destruction of the nucleus raphe obscurus and potentiation of serotonin-mediated behaviors following administration of the neurotoxin 3-acetylpyridine

Systemic administration of the neurotoxin 3-acetylpyridine (3-AP) to rats produced spontaneous episodes of spasmodic movement involving the trunk and limbs including torticollis, contortions of the trunk and rigid extension of the limbs. Because the neurotransmitter serotonin (5-HT) has been implicated in various human involuntary movement disorders, the functional and anatomical integrity of the 5-HT system in rats treated with 3-AP were examined. 5-HT-containing neurons in the brain stem were studied using immunohistochemical labeling with antiserum to 5-HT. Cells in the nucleus raphe obscurus were found to be altered following 3-AP treatment as shown by a decrease in 5-HT immunoreactivity as compared to control rats. No changes in 5-HT immunoreactivity were observed in any other region containing 5-HT cell bodies. Behaviorally, rats treated with 3-AP were 2.5-fold more sensitive to the ability of the 5-HT_1A agonist 8-OH-2-(di-n-propylamino)tetralin (8-OH-DPAT; 0.33–3.3 mg/kg) to produce the 5-HT syndrome. Similarly, 3-AP-treated rats were 2-fold more sensitive to the selective 5-HT₂ agonist 1-(2,5-dimethoxy-4-bromophenyl)-2-aminopropane (DOB; 0–1.0 mg/kg) at producing the head shake response. Although these behaviors associated with brain stem 5-HT receptors were potentiated by 3-AP, the hypothermic effect of 8-OH-DPAT which involves ascending mesencephalic 5-HT neurons was unchanged following 3-AP treatment. Treatment with 3-AP did not produce significant alterations of 5-HT or 5-hydroxyindoleacetic acid (5-HIAA) content in any brain region studied. Quantitative autoradiographic analysis of the density of 5-HT_1A receptors labeled with [³H]8-OH-DPAT revealed that these sites were largely unchanged in regions of the brain (frontal cortex, hippocampus and brain stem) and in the spinal cord. Similarly, few changes in the density of 5-HT₂ receptors measured with [³H]ketanserin were observed in various brain regions. These results suggest that neurons from the nucleus raphe obscurus are involved in the elicitation of 5-HT-mediated behavioral resonses by 5-HT_1A and 5-HT₂ receptor agonists that are thought to be mediated through brain stem and spinal cord mechanisms. In addition, because of the close neuroanatomical relationship of the nucleus raphe obscurus with various brain regions known to be involved in motor control, the destruction of this region by 3-AP may contribute to the spasmodic motor behaviors observed following 3-AP treatment.     

Protection against hippocampal kainate excitotoxicity by intracerebral administration of an adenosine A2A receptor antagonist

We have previously shown that the peripheral administration of an A_2A receptor agonist 2-p-(2-carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine hydrochloride (CGS 21680) protected the hippocampus against kainate-induced excitotoxicity. The present study utilises the intrahippocampal route to further investigate CGS 21680-mediated protection as well as examining the role of adenosine and both A₁ and A_2A receptors in kainate-induced excitotoxicity. Injections were made directly into the hippocampus of anaesthetised male Wistar rats. Following surgery and the administration of 0.25 nmol kainate in 1 μl of solution, the animals were left to recover for seven days before perfusion and brain slicing. Haematoxylin and eosin staining revealed substantial damage to the CA3 region. Co-administration of the A_2A receptor agonist CGS 21680 over a range of doses did not protect the region to any degree. Similarly neither the A₁ receptor agonist R-phenylisopropyladenosine (R-PIA), nor adenosine itself reduced kainate-induced damage. The intrahippocampal injection of the selective A_2A receptor antagonist, 4-(2-[7-amino-2-{2-furyl}{1,2,4}triazolo{2,3-a}{1,3,5}triazin-5-yl-amino]ethyl)phenol (ZM241385) however, significantly decreased kainate damage to the CA3 region. These results show that adenosine A_2A receptor-induced protection is most likely to be mediated peripherally and is probably not due to activation of A_2A receptors within the hippocampus. The lack of protection observed with either R-PIA or adenosine may be due to an inhibitory action of the A_2A receptor on the neuroprotective A₁ receptor. Importantly, this study also questions the role of endogenously released adenosine in protecting the hippocampus from excitotoxic damage.     

Effects of serotonergic drugs in experimental brain ischemia: evidence for a protective role of serotonin in cerebral ischemia

We have examined the significance of the serotonergic system in the pathophysiology of ischemic brain damage. Permanent occlusion of the middle cerebral artery (MCA) was performed in male NMRI mice. After 48 h, the animals received a transcardiac injection of carbon black. The area of ischemia was restricted to the neocortex and its size was determined planimetrically by means of an image analyzing system. In control experiments, the NMDA antagonist dizocilpine (MK-801), the AMPA/kainate antagonist NBQX (2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(F)-quinoxaline) and the L-type calcium channel blocker nimodipine all produced a significant reduction in ischemic injury of the mouse neocortex. Interestingly, all of the 5-HT_1A agonists tested (ipsapirone, CM 57493 [4-(3-trifluoromethylphenyl)-1-(2-cyanoethyl)-1,2,3,6-tetrahydropyridine] and urapidil) were equally efficacious in reducing ischemic injury. On the other hand, the 5-HT₂ antagonist naftidrofuryl failed to protect the brain tissue significantly against ischemic brain damage. Roxindole, a 5-HT_1A agonist and 5-HT uptake inhibitor, was the most potent serotonergic compound tested. In order to examine the effects of 5-HT_1A receptor activation in a different context, 10 min of forebrain ischemia was induced in male Wistar rats by a bilateral occlusion of the common carotid arteries combined with systemic hypotension. Administration of the 5-HT_1A agonist CM 57493 reduced the neuronal damage within the ventral hippocampus and the entorhinal cortex as assessed histologically 7 days after ischemia. Finally, we found that 5-HT_1A agonists are capable of reducing neuronal damage of cultured neocortical and hippocampal neurons subjected to a chemical hypoxia or glutamate in a dose dependent manner. These data suggest that 5-HT, released during ischemia, may have protective effects in the pathophysiology of ischemic brain damage through a direct action on neurons mediated via the inhibitory 5-HT_1A receptor subtype. The results obtained from different in vivo and in vitro models indicate that 5-HT_1A agonists are promising agents for the treatment of ischemic brain disorders.     

A reliable method for the preferential activation of C- or A-fibre heat nociceptors

The thyroid hormone triiodothyronine (T3) augments and accelerates the effects of antidepressant drugs. Although the majority of studies showing this have used tricyclics, a few studies have shown similar effects with the selective serotonin re-uptake inhibitor (SSRI) fluoxetine. In this study we investigated the effects of fluoxetine (5 mg/kg), T3 (20 μg/kg) and the combination of these drugs, each administered daily for 7 days, on serotonergic function in the rat brain, using in vivo microdialysis. Fluoxetine alone induced a trend towards desensitization of 5-HT_1A autoreceptors as shown by a reduction in the effect of 8-OH-DPAT to lower 5-HT levels in frontal cortex, and desensitized 5-HT_1B autoreceptors in frontal cortex. The combination of fluoxetine and T3 induced desensitization of 5-HT_1B autoreceptors in hypothalamus. Since there is evidence linking hypothalamic function and depression, we suggest that this effect may partly account for the therapeutic efficacy of the combination of an SSRI and T3.