Effects of the beta3-adrenoceptor (Adrb3) agonist SR58611A (amibegron) on serotonergic and noradrenergic transmission in the rodent: relevance to its antidepressant/anxiolytic-like profile.

SR58611A is a selective beta(3)-adrenoceptor (Adrb3) agonist which has demonstrated antidepressant and anxiolytic properties in rodents. The present study confirmed the detection of Adrb3 mRNA transcript in rodent brain sub-regions and evaluated the effect of SR58611A on serotonergic and noradrenergic transmission in rats and mice in an attempt to elucidate the mechanism(s) underlying these properties. SR58611A (3 and 10 mg/kg, p.o.) increased the synthesis of 5-HT and tryptophan (Trp) levels in several rodent brain areas (cortex, hippocampus, hypothalamus, striatum). Moreover, SR58611A (10 mg/kg, p.o.) increased the release of 5-HT assessed by in vivo microdialysis in rat prefrontal cortex. Systemic (3 mg/kg, i.v.) or chronic administration of SR58611A (10 mg/kg, p.o.), in contrast to fluoxetine (15 mg/kg, p.o.), did not modify the activity of serotonergic neurons in the rat dorsal raphe nucleus. The increase in 5-HT synthesis induced by SR58611A was not observed in Adrb3s knockout mice, suggesting a selective involvement of Adrb3s in this effect. SR58611A (3 and 10 mg/kg, p.o.) did not modify norepinephrine synthesis and metabolism but increased its release in rat brain. Repeated administration of SR58611A (10 mg/kg, p.o.) did not modify basal norepinephrine release in rat prefrontal cortex whereas it prevented its tail-pinch stress-induced enhancement similarly to reboxetine (15 mg/kg, p.o.). Finally SR58611A increased the firing rate of noradrenergic neurons in the rat locus coeruleus following systemic (3 mg/kg, i.v.) or local (0.01 and 1 microM) but not chronic (10 mg/kg, p.o.) administration. These results suggest that the anxiolytic- and antidepressant-like activities of SR58611A involve an increase of brain serotonergic and noradrenergic neurotransmissions, triggered by activation of Adrb3s.

Effects of the vasopressin (V1b) receptor antagonist, SSR149415, and the corticotropin-releasing factor 1 receptor antagonist, SSR125543, on FG 7142-induced increase in acetylcholine and norepinephrine release in the rat.

Arginine vasopressin and corticotropin-releasing factor are two neuroactive peptides that regulate hypothalamic-pituitary-axis and associated stress response. While the potential antidepressant and anxiolytic profiles of corticotropin-releasing factor 1 antagonists have been well studied, the concept of blockade of vasopressin system as another approach for the treatment of emotional processes has only been made available recently by the synthesis of the first non-peptide antagonist at the V1b receptor, SSR149415. In the present study SSR149415 has been compared with the corticotropin-releasing factor 1 antagonist SSR125543 and with anxiolytic and antidepressant drugs on the response of hippocampal cholinergic and cortical noradrenergic systems to the anxiogenic benzodiazepine receptor inverse agonist FG 7142. Acute (0.3-10 mg/kg, i.p.) and long-term administration (10 mg/kg, i.p., 21 days) of SSR149415 and SSR125543 reduced the FG 7142-induced increase in extracellular concentrations of acetylcholine in the hippocampus of anesthetized rats measured by microdialysis. By contrast acute and long-term administration of SSR149415 failed to reduce the FG 7142-induced increase in the release of norepinephrine in the cortex of freely moving rats. The present results demonstrate that the two compounds have similar profiles in a model of activation by an anxiogenic drug of the hippocampal cholinergic system and they suggest that SSR149415 and SSR125543 may have anti-stress anxiolytic and antidepressant effects via a mechanism of action different from classical benzodiazepine ligands and noradrenergic antidepressants.

Control by tachykinin NK(2) receptors of CRF(1) receptor-mediated activation of hippocampal acetylcholine release in the rat and guinea-pig.

In vivo microdialysis was employed to explore the effects of different selective non-peptides NK(1),NK(2) and NK(3) receptor antagonists on the corticotropin releasing factor (CRF)-induced release of acetylcholine (ACh) in the hippocampus of rats and guinea-pigs. In both species, the intracerebroventricular (i.c.v.) administration of CRF produced a time- and dose-dependent increase in hippocampal ACh release that was totally suppressed by an intraperitoneally (i.p.) pretreatment with the selective non-peptide CRF(1) receptor antagonist antalarmin (30 mg/kg). Pretreatment with the selective NK(2) receptor antagonist SR48968 (1mg/kg, i.p.) significantly reduced the increase of ACh induced by CRF. In contrast, its low-affinity enantiomer SR48965 (1mg/kg, i.p.) or the NK(1) receptor antagonist, GR205171 (1mg/kg, i.p.) did not exert any antagonist effect. Moreover, administration of the selective NK(3) receptor antagonist SR142801 (1mg/kg, i.p.) did not significantly reduce the CRF-induced hippocampal ACh release in guinea-pigs (the only species studied). The selective activity of SR48968 versus GR205171 or SR142801 indicates that NK(2) receptors play a major role in the control of CRF-induced hippocampal ACh release. Moreover, in freely moving rats, two sessions of stroking of the neck and back of the rat for 30 min, at 90 min intervals, known to be a stressful stimulus, produced a marked and reproducible increase in hippocampal ACh release. This effect was prevented by the administration of the two selective non-peptide CRF1 and NK(2) receptor antagonists antalarmin (30 mg/kg, i.p.) and SR48968 (1mg/kg, i.p.), respectively. This suggests that stress-induced activation of the hippocampal ACh system may be under the control of both endogenously released CRF and NKA, and opens the possibility of the existence of a functional interplay between the pathways containing these peptides as we observed in our experiments on anaesthetized animals.

The preclinical pharmacologic profile of tiapride.

Tiapride is a benzamide derivative that has been used successfully in the clinic for a number of years for the treatment of agitation and aggressiveness in elderly patients. Like many substituted benzamides, tiapride specifically blocks dopamine receptors in the brain. It has affinity for dopamine D(2) (IC(50) = 110-320 nM) and D(3) (IC(50) = 180 nM) receptors in vitro but lacks affinity for dopamine D(1) and D(4) receptors and for non-dopaminergic receptors including H(1), alpha(1), alpha(2)-adrenergic and serotonergic receptors. Tiapride also shows dose-related inhibition of [3H]-raclopride binding in limbic areas and in the striatum of the rat in vivo (ED(50) approximately 20 mg/kg, ip). In microdialysis experiments, tiapride (over the range 10-30 mg/kg, ip) increased extracellular levels of dopamine in the nucleus accumbens and striatum, a reflection of its blockade of postsynaptic dopamine receptors in these brain areas. In behavioral experiments in rats, lower doses of tiapride (ED(50) = 10 mg/kg, ip) antagonised dopamine agonist-induced hyperactivity while higher doses (ED(50) = 60 mg/kg, ip) were required to block stereotyped movements. In addition, doses of tiapride up to 200 mg/kg, ip failed to induce catalepsy, an effect observed with many other drugs which block dopamine receptors. In tests of conditioned behavior in rats, tiapride was found to give rise to an interoceptive stimulus associated with dopamine receptor blockade at doses (ED(50) = 2.2 mg/kg, ip) much lower than those producing motor disturbances or sedation (ED(50) = 40 mg/kg, ip), in striking contrast to a range of conventional or atypical neuroleptics that produced interoceptive stimulus and sedation at similar doses. Furthermore, the acquisition by rats of a place-learning task in a water maze was not affected by tiapride (over the range 3-30 mg/kg, ip), whereas haloperidol (MED = 0.25 mg/kg, ip) and risperidone (MED = 0.03 mg/kg, ip) impaired performance. The preclinical pharmacologic and behavioral profile of tiapride suggests that its clinical activity may be due to a selective blockade of dopamine D(2) and D(3) receptors in limbic brain regions. The results are also consistent with a lack of motor or cognitive side effects.

Neurochemical characteristics of amisulpride, an atypical dopamine D2/D3 receptor antagonist with both presynaptic and limbic selectivity.

The benzamide derivative amisulpride shows a unique therapeutic profile being antipsychotic, at high doses, and disinhibitory, at low doses, while giving rise to only a low incidence of extrapyramidal side effects. In vitro, amisulpride has high affinity and selectivity for the human dopamine D2 (Ki = 2.8 nM) and D3 (Ki = 3.2 nM) receptors. Amisulpride shows antagonist properties toward D3 and both pre- and postsynaptic D2-like dopamine receptors of the rat striatum or nucleus accumbens in vitro. At low doses (< or = 10 mg/kg) amisulpride preferentially blocks presynaptic dopamine autoreceptors that control dopamine synthesis and release in the rat, whereas at higher doses (40-80 mg/kg) postsynaptic dopamine D2 receptor occupancy and antagonism is apparent. In contrast, haloperidol is active in all of these paradigms within the same dose range. Amisulpride preferentially inhibits in vivo binding of the D2/D3 antagonist [3H]raclopride to the limbic system (ID50 = 17 mg/kg) in comparison to the striatum (ID50 = 44 mg/kg) of the rat, increases striatal and limbic tissue 3,4-dihydroxyphenylacetic acid levels with similar potency and efficacy, and preferentially increases extracellular 3,4-dihydroxyphenylacetic acid levels in the nucleus accumbens when compared to the striatum. Haloperidol shows similar potency for the displacement of in vivo [3H]raclopride binding in striatal and limbic regions and preferentially increases striatal tissue 3,4-dihydroxyphenylacetic acid levels. The present data characterize amisulpride as a specific dopamine receptor antagonist with high and similar affinity for the dopamine D2 and D3 receptor. In vivo, it displays a degree of limbic selectivity and a preferential effect, at low doses, on dopamine D2/D3 autoreceptors. This atypical profile may explain the therapeutic efficacy of amisulpride in the treatment of both positive and negative symptoms of schizophrenia.

DTG-induced circling behaviour in rats may involve the interaction between sigma sites and nigro-striatal dopaminergic pathways.

The distribution of sigma sites in brain areas enriched in dopamine, and the finding that circling behaviour can be elicited by specific sigma ligands such as DTG (di-o-tolylguanidine) suggest a modulatory role of these sites in the dopaminergic system. The present study was carried out to investigate further this hypothesis. Circling behaviour induced in rats by unilateral intranigral injection of DTG (10 nmol/rat) was decreased by haloperidol (0.1 mg/kg, i.p.), clebopride (0.25 mg/kg, i.p.) and SCH 23390 (0.03 mg/kg, s.c.) indicating that an interaction between sigma sites and the midbrain dopaminergic system may be involved in this rotational behaviour. Microdialysis experiments in freely moving rats showed that unilateral intranigral injection of DTG (5, 10, 20 nmol/rat) produced increases in extracellular levels of dopamine metabolites (DOPAC, HVA) in the ipsilateral striatum which correlated with the number of rotations. In addition intranigral injection of DTG (10 nmol/rat) produced increases in tissular dopamine metabolite levels in the ipsilateral striatum without affecting dopamine metabolite levels in limbic structures. These results indicate that sigma sites may be involved in the modulation of the dopaminergic motor system.

Alpha 1-adrenoceptor antagonists differentially control serotonin release in the hippocampus and striatum: a microdialysis study.

Using the in vivo microdialysis technique, we have studied the effect of the systemic administration of several alpha 1-adrenoceptor antagonists on the extracellular levels of serotonin (5-HT) in the rat hippocampus. Prazosin, and to a lesser extent, terazosin, decreased these levels by 50-65% for 0.03-0.4 mg/kg, i.v. and by 30-40% for 0.1-0.4 mg/kg, i.v., respectively. In contrast, alfuzosin, an alpha 1-adrenoceptor antagonist with poor brain penetration, did not significantly affect these levels even at the high dose of 0.4 mg/kg, i.v. When perfused into the hippocampus through the dialysis probe, prazosin (1-10 microM) induced a more limited (20-30%) and delayed decrease in 5-HT outflow. These results support the existence of a central noradrenergic facilitatory influence, mediated by alpha 1-adrenoceptors, on serotonergic neurons projecting to the hippocampus. In the striatum prazosin (0.4 mg/kg, i.v.) decreased 5-HT levels to a smaller extent (-35%) than in the hippocampus (-65%), suggesting the existence of differences in the degree of noradrenergic influence on median and dorsal raphé nuclei, which preferentially project to the hippocampus and striatum, respectively.

Muscarinic receptor-mediated increase in extracellular inositol 1,4,5-trisphosphate levels in the rat hippocampus: an in vivo microdialysis study.

The extracellular concentration of inositol 1,4,5-trisphosphate (IP3) has been monitored in the ventral hippocampus of the anesthetized rat by using a microdialysis technique coupled to a radioreceptor assay. Three hours after the implantation of the cannula, basal extracellular concentration of IP3 (corrected for a 9% recovery) was 71 nM (0.39 pmol/60-microliters fraction) and remained stable for at least 5 h. Local infusion of carbachol for 60 min caused a significant concentration-related increase in extracellular IP3 levels (0, 24, and 57% at 1, 50, and 100 microM, respectively). Acetylcholine (100 microM) and muscarine (100 microM) increased IP3 outflow by 40 and 42%, respectively. The effect of carbachol was fully prevented by coinfusion of 10 microM pirenzepine and reduced by 1 microM tetrodotoxin indicating that the carbachol response is mediated by neuronal muscarinic receptors. These data demonstrate the feasibility of using microdialysis and a radioreceptor assay to measure IP3 in the extracellular space. This approach could prove useful for the study of the in vivo operation of muscarinic and, by extension, a number of receptors coupled to phosphoinositide turnover.

Effect of local injection of 8-OH-DPAT into the dorsal or median raphe nuclei on extracellular levels of serotonin in serotonergic projection areas in the rat brain.

Using in vivo microdialysis, we have examined the effects of local administration of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) into either the dorsal (DRN) or the median (MRN) raphe nucleus on extracellular levels of serotonin (5-HT) in the corresponding projection fields, namely striatum and ventral hippocampus in the anaesthetized rat. Local injection of 8-OH-DPAT (0.5 microgram/0.1 microliter) in the DRN reduced extracellular 5-HT levels in the striatum (-55%) and to a lesser extent in the hippocampus (-22%). When injected at the same dose in the MRN, 8-OH-DPAT caused a marked decrease in 5-HT output in the hippocampus (-41%) and had no effect in the striatum. Autoradiographic studies performed on brains from animals that received a local injection of [8H]8-OH-DPAT into either the DRN or MRN under similar experimental conditions indicated that the radioactivity remained localized within each midbrain raphe nucleus. These results confirm anatomical data demonstrating that the striatum and the ventral hippocampus receive their serotonergic innervation preferentially from the DRN and the MRN, respectively.

Effect of the putative 5-HT1A receptor antagonist NAN-190 on rat brain serotonergic transmission.

Based on the fact that 1-(2-methoxyphenyl)-4(4-(2-phtalimido)butyl)piperazine (NAN-190), a high-affinity ligand for 5-HT1A and alpha 1-adrenoceptors, antagonizes the behavioural effects of the 5-HT1A receptor agonist 8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin), it has been suggested that this drug behaves as a 5-HT1A receptor antagonist. In the present study we examined the effects of this putative 5-HT1A receptor antagonist on rat brain serotonergic neurotransmission. In hippocampal slices of immature rats, NAN-190 but not prazosin potently antagonized (IC50 = 29 nM) the inhibitory effect of 8-OH-DPAT (1 microM) on carbachol-stimulated phosphoinositide turnover but (up to 1 microM) failed to alter the carbachol response. Similarly, NAN-190 (0.1 microM) almost totally prevented the inhibition by 8-OH-DPAT (1 microM) of forskolin-stimulated adenylate cyclase activity in adult rat hippocampal slices but, per se, was without effect on the forskolin response. These results indicate that NAN-190 is a potent antagonist at postsynaptic 5-HT1A receptors in vitro. However, NAN-190 also potently antagonized (IC50 = 0.16 nM) the stimulation by norepinephrine of phosphoinositide turnover in rat cortical slices. In this respect NAN-190 was a 250-fold more potent antagonist than prazosin (IC50 = 49 nM). Thus, in addition to its 5-HT1A receptor antagonist properties, NAN-190 has potent alpha 1-adrenoceptor blocking properties.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence for differing origins of the serotonergic innervation of major cerebral arteries and small pial vessels in the rat.

We have studied the nature and origin of the serotonergic innervation of two distinct anatomical cerebrovascular compartments, namely, small pial vessels and major cerebral arteries, in the rat. To this end, the levels of serotonin [5-hydroxytryptamine (5-HT)] and 5-hydroxyindoleacetic acid (5-HIAA) were measured by HPLC in both cerebrovascular compartments after either bilateral sympathectomy or destruction of the ascending serotonergic pathways, which originate from the raphe nuclei. We first showed that the small pial vessel samples were not contaminated by underlying cortical tissues through the use of an immunohistochemical approach that revealed the glia limitans, the most superficial cortical layer. Superior cervical ganglionectomy caused a marked decrease in noradrenaline concentrations in major cerebral arteries (-77%), although the reduction was less pronounced (-34%) in small pial vessels. Sympathectomy decreased by 33% 5-HT concentrations in the major cerebral arteries but was without effect on 5-HT levels in the small pial vessels. Destruction of the ascending serotonergic pathways (via local administration of 5,7-dihydroxytryptamine into the ventral tegmental area) produced a dramatic fall in 5-HT and 5-HIAA concentrations in both vascular compartments. To establish the authenticity of the serotonergic innervation, the synthesis of 5-HT [as assessed by measuring the accumulation of 5-hydroxytryptophan (5-HTP) after decarboxylase inhibition] was measured in the two vascular beds under control conditions and after destruction of the ascending serotonergic pathways. The rate of accumulation of 5-HTP was higher in the small pial vessels than in major cerebral arteries, an observation that indicates an important de novo synthesis of 5-HT in small pial vessels.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential effects of electrical stimulation of the dorsal raphe nucleus and of cervical sympathectomy on serotonin and noradrenaline concentrations in major cerebral arteries and pial vessels in the rat.

The levels of noradrenaline (NA), serotonin (5-HT), and 5-hydroxyindoleacetic acid were measured by HPLC and compared between the large arteries of the circle of Willis and the small pial vessels in the rat, following either electrical stimulation of the dorsal raphe nucleus or bilateral superior cervical ganglionectomy. With electrical stimulation, the 5-HT concentrations were reduced (-48%) in the small pial vessels, but were unchanged in the major cerebral arteries. NA concentrations were dramatically reduced following cervical sympathectomy in the large arteries (-77%), though the reduction was less pronounced (-34%) in the small vessels. Sympathectomy caused a significant decrease in the 5-HT concentration of the major cerebral arteries (-33%), but was without effect on the 5-HT levels of the small pial vessels. These results show that an appreciable fraction of the perivascular 5-HT measured in the small pial and the large cerebral arteries originates from different sources.

Pharmacological characterization of serotonin-stimulated phosphoinositide turnover in brain regions of the immature rat.

The effects of serotonin (5-HT) and related agonists and antagonists on phosphoinositide turnover have been investigated in several brain regions of the immature rat. In the presence of LiCl, 5-HT caused a marked increase in total [3H]inositol phosphate levels in cortical (maximal effect + 420%, EC50 = 7 microM) and to a lesser extent in hippocampal and striatal slices prepared from the immature (8-day-old) rat; the cortical 5-HT-induced phosphoinositide response was tetrodotoxin resistant. The magnitude of the increase in the cortical phosphoinositide response caused by 5-HT was maximal at 1 day postnatal and progressively declined to reach 6% of this maximal response in the adult. After incubation of immature (8-day-old) rat cortical slices for 2.5 min with 5-HT (in the absence of LiCl), inositol 1-phosphate, inositol 1,4-bisphosphate, inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate levels increased about 2-fold. A variety of 5-HT2 or mixed 5-HT1/5-HT2 agonists stimulated total [3H]inositol phosphate formation in the immature rat cortex and hippocampus with a rank order of potency [alpha(+)-methyl-5-HT greater than quipazine greater than MK 212 greater than 5-HT] which resembles their potencies at the 5-HT2 binding site. In contrast, the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin, the 5-HT1B agonists 1-(m-trifluoromethylphenyl)piperazine and 1-(m-chlorophenyl)-piperazine and the 5-HT3 agonist 2-methyl-5-HT were inactive.(ABSTRACT TRUNCATED AT 250 WORDS)

Concentrations of putative neurovascular transmitters in major cerebral arteries and small pial vessels of various species.

The levels of noradrenaline, neuropeptide Y, 5-hydroxytryptamine, and substance P were measured and compared between the large arteries of the circle of Willis and the small cerebral vessels of the pia mater in the rat, rabbit, cat, and monkey. In all species, noradrenaline and neuropeptide Y concentrations were greater in the larger arteries than in small pial vessels. Noradrenaline concentrations were dramatically reduced following cervical sympathectomy, with the extent of diminution differing greatly in the various species; the effects of cervical ganglionectomy on neuropeptide Y concentrations were less pronounced. 5-Hydroxytryptamine concentrations in rats, cats, and rabbits were significantly greater in the small pial vessels, although measurable concentrations existed in the circle of Willis. In cats and monkeys, substance P was found in major arteries, but was not detectable at the level of the small pial vessels. The differences in the regional distribution of the various neurotransmitter candidates in the cerebrovascular bed may reflect their physiological significance.

Lack of correlation between plasma DOPEG and urinary MOPEG levels in depressed patients.

Twenty-four-hour urinary excretion of 3-methoxy,4-hydroxyphenylethyleneglycol (MOPEG) and levels of free and conjugated plasma 3,4-dihydroxyphenylethyleneglycol (DOPEG) were measured in 56 depressed patients to find a possible correlation between these two peripheral indices of cerebral noradrenergic activity. Plasma DOPEG was measured at 9:00 AM on the same day that urine was collected for the measurement of MOPEG. All depressed patients were diagnosed as having affective disorders according to DSM-III. No correlation was found between plasma free or conjugated DOPEG levels and urinary MOPEG output. This lack of correlation was found in the total sample of depressed patients (56), in 45 patients diagnosed as having major depressive episodes, and in 24 depressed patients diagnosed as major depressive with melancholia. The authors discuss the significance of this lack of correlation between two peripheral indices of central noradrenergic metabolism.

Reduction of cortical dopamine, noradrenaline, serotonin and their metabolites in Parkinson's disease.

Dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, noradrenaline, serotonin and 5-hydroxyindoleacetic acid concentrations were measured in several cortical areas, hippocampus and, for comparison, in the caudate nucleus, from control subjects and parkinsonian patients. Substantial amounts of these compounds were detected in hippocampus, and entorhinal, cingulate and frontal cerebral cortices of control subjects. In patients who had discontinued L-DOPA at least 4 days before death (group I), the levels of dopamine and its metabolites were reduced in these cortical areas, although to a lesser extent than in the caudate nucleus. In patients on continuous L-DOPA treatment (i.e. having received the last dose of L-DOPA 0-24 h before death, group II), cortical dopamine levels were less reduced than in group I patients and dopamine metabolite levels were similar to those of controls. The ratio of the concentrations of homovanillic acid to dopamine was increased in the caudate nucleus and entorhinal cortex but not in the other cortical areas of group I parkinsonian patients. Cortical noradrenaline concentrations were also diminished, the decrement being similar in groups I and II. A reduction of serotonin and its metabolite in the caudate nucleus and hippocampus and a diminution of serotonin levels in the frontal cortex were observed in group I patients. In these patients, the 5-hydroxyindoleacetic acid to serotonin ratio was increased in the caudate nucleus and frontal cortex but not in the other cortical areas. The results are discussed in relation to the pathophysiology of the psychiatric and cognitive disturbances observed in some parkinsonian patients.

Dopamine deficiency in the cerebral cortex in Parkinson disease.

We measured the concentrations of dopamine, 3,4-dihydroxyphenylacetic acid, and homovanillic acid in several cortical areas from controls and parkinsonian patients. Substantial amounts of dopamine and its metabolites were detected in hippocampus and entorhinal, cingulate, and frontal cortex of controls. In parkinsonian patients the levels of dopamine and its metabolites were reduced in these neocortical areas and hippocampus. Diminution of cortical dopaminergic transmission may play a role in the mental impairment of some Parkinson patients.

Differential effects of inescapable footshocks and of stimuli previously paired with inescapable footshocks on dopamine turnover in cortical and limbic areas of the rat.

The effect of electric footshocks and of exposure to environmental stimuli paired with electrical shocks upon the dopaminergic activity in various cortical and limbic areas of the rat were evaluated by measuring dihydroxyphenylacetic acid (DOPAC) levels in these areas. In animals exposed to a 20 min electric footshock session DOPAC concentrations were significantly increased in the antero-medial and sulcal frontal cortices, olfactory tubercle, nucleus accumbens and amygdaloid complex (by 66, 37, 28, 55 and 90% respectively). Re-exposure of rats to an environment where they had been shocked 24 h earlier induced an elevation of DOPAC content only in the anteromedial frontal cortex (by 47%). Plasma corticosterone levels were elevated in both situations. No change in serotonin or 5-hydroxyindolacetic acid content of these areas could be detected in either situation. The results show that electric footshocks and environmental stimuli associated to previous shocks both activate central dopaminergic systems, although the patterns of activation are different.

Determination of 5-hydroxytryptophan, serotonin and 5-hydroxyindoleacetic acid in rat and human brain and biological fluids by reversed-phase high-performance liquid chromatography with electrochemical detection.

A rapid and sensitive method for the concurrent determination of 5-hydroxytryptophan, serotonin and 5-hydroxyindole acetic acid by reversed-phase high-performance liquid chromatography with electrochemical detection has been developed. The separation of the indolic compounds was achieved using a phosphate-citric acid eluent containing 5% methanol. Detection limits in the low picogram range were found. The method has been applied to the determination of the indolic compounds in rat and human brain tissues, as well as in human plasma and cerebrospinal fluid. Tissue and plasma preparation required only deproteinization before chromatography, while cerebrospinal fluid was directly applied to the column.