DSP4-induced two-way active avoidance impairment in rats: Involvement of central and not peripheral noradrenaline depletion

N-2-chloroethyl-N-ethyl-2-bromobenzylamine (DSP4) (50 mg/kg IP), a new selective noradrenaline (NA) neurotoxin, was found to cause a severe impairment of the acquisition of a two-way active avoidance task 1 week following adminsitration, an effect that was blocked by the selective NA uptake inhibitor desipramine. In a second experiment, systemically injected 6-OHDA (2×30 mg/kg IP) was not found to cause any avoidance impairment, althogh its effects upon peripheral NA were still evident 21 days after administration: The peripheral NA depletion caused by DSP4 almost disappeared 14 days after injection. In a third experiment, the avoidance impairment induced by DSP4 was produced even 10 weeks after treatment. Data from both the shuttlebox experiment and an activity box experiment suggest that the acquisition impairment is not readily explained on the basis of some deficit in spontaneous behavior or an altered perception of pain. The present data suggest that the effect of DSP4 upon active avoidance asquisition is mediated via central, and not peripheral NA neurons.     

Selective brain noradrenaline depletion induced by the neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP 4) does not prevent the memory facilitation induced by a muscarinic agonist in mice

These experiments investigated the effects of central noradrenaline (NA) depletion and its interaction with cholinergic and dopaminergic mechanisms upon retention of a passive-avoidance response in mice. The NA selective neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP 4) (50 mg/kg IP, 7 days) was injected into mice to produce depletion of NA in frontal cortex, hypothalamus, cerebellum, midbrain and brain stem without any significant change in dopamine (DA) levels in frontal cortex, striatum, hypothalamus and midbrain. Depletion of brain NA produced by DSP 4 was significantly but not completely prevented by the NA uptake inhibitor desmethylimipramine (DMI) (10 mg/kg IP, 30 min before DSP 4 injection). Despite the marked NA depletion, DSP 4 neither impaired the retention of a passive-avoidance response in mice nor prevented the enhancement of retention of this response induced by the central muscarinic agonist oxotremorine (OTM) (0.05 mg/kg IP, immediately after training. This lack of effect of DSP 4 on retention was prevented neither by DMI nor by the serotonin uptake inhibitor fluoxetine (5 mg/kg IP, 30 min before DSP 4 injection). The enhancement of retention induced by OTM in the groups of mice injected with either water or DSP 4 was prevented by atropine (0.5 mg/kg IP, 20 min before training) but not by methylatropine in the same experimental conditions. This suggests that both in controls and DSP 4-pretreated mice, the primary effect of OTM is due to an interaction with muscarinic brain receptors.(ABSTRACT TRUNCATED AT 250 WORDS)     

Limited involvement of central noradrenergic pathways in morphine-induced antinociception

Lesioning of cerebral and spinal noradrenergic terminals by the neurotoxin DSP4 (50 mg/kg, 7 days prior to testing) significantly attenuated the effect of morphine (2.5–7.5 mg/kg) in rats tested with the hot-plate test. The effect of DSP4 was prevented by pretreatment with the selective inhibitor of uptake of NA, desipramine. Treatment with DSP4 did not attenuate the effect of morphine (5 mg/kg) in the flinch-jump and tail-flick tests, and did not by itself change the nociceptive thresholds in any of the tests. It is concluded that noradrenergic structures in the central nervous system play a limited role in analgesia induced by morphine.     

Central noradrenaline depletion antagonizes aspects ofd-amphetamine-induced hyperactivity in the rat

The effects of noradrenaline (NA) depletion upon amphetamine-induced hyperactivity were examined in five experiments. Central NA depletion via either systemic DSP4 or neonatal 6-OHDA antagonised the amphetamine-induced (2 mg/kg SC) increase in rearing behaviour, whereas lesions of the dorsal noradrenergic bundle using 6-hydroxydopamine antagonised the increase in locomotor activity. Peripheral NA depletion following systemic 6-hydroxydopamine to adult rats did not cause any changes in motor activity after acute amphetamine administration. Desipramine, the selective NA uptake inhibitor, blocked the effects of DSP4 upon amphetamine-induced rearing. NA depletion antagonised hyperactivity produced by the 2 mg/kg dose of amphetamine, but not the hyperactivity (rearing or locomotion) effects of amphetamine at 1, 4 or 8 mg/kg.     

6-OHDA lesion to the dorsal noradrenergic bundle alters morphine-induced locomotor activity and catalepsy

Rats show an initial depression in locomotor activity in response to doses of morphine greater than 5 mg/kg during the first hour after injection which is followed by a prolonged hyperactive phase. The effect of bilateral 6-hydroxydopamine (6-OHDA) lesions to the dorsal noradrenergic bundle on this biphasic action of morphine was studied. These lesions were found to significantly potentiate the locomotor depressant effects of morphine at 10.0 and 20.0 mg/kg while leaving the subsequent stimulant action of morphine unchanged. The cataleptic action of morphine at 20.0 mg/kg as measured in a separate test was also potentiated. These lesions were found to deplete hippocampal and cortical noradrenaline (NA) to 3% and hypothalamic NA to 32% of control values and also to cause significant increases in cerebellar and spinal NA. These data suggest a role for NA in the depressant effects of morphine but not in its subsequent stimulant actions which appear to be mediated by other neurochemical systems.     

Separation of the associative and non-associative effects of brain serotonin released by p-chloroamphetamine: Dissociable serotoninergic involvement in avoidance learning,pain and motor function

p-Chloramphetamine (PCA, 0.63–5 mg/kg IP) injected 30–60 min before testing produced a dose-related impairment of avoidance acquisition, prolonged reaction time in the hot-plate test and increased locomotor activity. Pretreatment with the selective serotonin (5-HT) uptake inhibitor zimeldine (10 mg/kg IP) blocked these behavioural effects. Degeneration of brain 5-HT neurons by a high neurotoxic dose of PCA (2×10 mg/kg IP) or inhibition of tryptophan hydroxylase by p-chlorophenylalanine (300 mg/kg IP) also blocked the behavioural effects of PCA. There was a complete blockade of the PCA-induced avoidance deficit following pretreatment with metergoline, a central 5-HT receptor blocking agent. On the other hand, metergoline failed to block the hot-plate analgesia and the increased locomotion caused by PCA. Depletion of brain NA and DA by the tyrosine hydroxylase inhibitor H44/68 did not counteract the PCA effect on avoidance or hot-plate performance, but reduced the locomotor stimulating effect. The selective NA neurotoxin DSP4 (50 mg/kg IP) or the opiate antagonist naloxone (1 mg/kg) failed to affect the PCA-induced modulations of the behaviours studied. In addition, PCA administration in doses that caused avoidance deficits, did not result in motor impairment as assessed by the tread mill test. The above results support the hypothesis that the PCA-induced impairment of active avoidance acquisition does not involve changes in nociception or altered locomotor activity. It is concluded that behavioural processes related to serotonergic neurotransmission can be independently modified, suggesting differences in the underlying 5-HT mechanisms.     

Discontinuation of chronic clonidine treatment: Evidence for facilitated brain noradrenergic neurotransmission

Summary Clonidine, 0.1 mg/kg i.p., or saline was administered to mice twice daily for 12 days. After withdrawal of the drug (14–18.5 h after the last injection), the accumulation of Dopa during 30 min after inhibition of central aromatic amino acid decarboxylase by NSD 1015 (3-hydroxy-benzyl hydrazine, 150 mg/kg i.p.) was significantly increased in the noradrenaline (NA)-rich, but dopamine (DA)-poor, brain stem but not in the DA-rich, but NA-poor corpus striatum. An increased Dopa accumulation was also found in the limbic system and, probably, in the hemispheres. The central accumulation of 5-hydroxytryptophan (5-HTP) was significantly increased in the limbic system. Clonidine, 0.1 mg/kg i.p., administered to mice during the withdrawal phase caused reduction of the accumulation of Dopa and 5-HTP (during 30 min after NSD 1015, 150 mg/kg i.p.) in all brain regions studied to approximately the same levels, as when given to saline-pretreated controls. The disappearance rates of brain NA and DA after synthesis inhibition (-methyl-p-tyrosine methylester HCl 250 mg/kg i.p., 4 h) were probably not altered by clonidine withdrawal.The locomotor stimulation by the central catecholamine receptor agonists apomorphine plus clonidine (1.5 mg/kg i.p. both) after reserpine (10 mg/kg, 4 h) pretreatment was significantly enhanced in clonidine withdrawal mice. However, the motor stimulation by apomorphine (1.5 mg/kg i.p.) alone after reserpine pretreatment was not affected. The spontaneous motor activity was not significantly altered by the withdrawal of clonidine.The data show that brain NA and 5-HT systems are affected by clonidine withdrawal in the opposite direction to that seen after acute administration of low doses of the drug, when they are inhibited. Since postsynaptic central -adrenoceptors, which are stimulated by high but not by low doses of clonidine, showed an enhanced response to high doses of clonidine following discontinuation of a chronic low dose regimen, supersensitivity due to functional noradrenergic denervation might be implicated.     

DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine)--a useful denervation tool for central and peripheral noradrenaline neurons

The effect of N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4) on monoamine neurons was studied in mice and rats. In mice DSP4 produced acutely a marked reduction of endogenous noradrenaline (NA), [3H]NA uptake and nerve density of the adrenergic nerves in the iris and atrium. Pronounced accumulations of NA were observed in non-terminal axons, which is a degenerative sign, while no changes were found in the NA cell bodies. A marked recovery of all parameters analysed was found as soon as 1 week after DSP4. In the mouse CNS, however, there was a marked and long-lasting NA reduction, especially in the cerebral and cerebellar cortex and spinal cord, leaving dopamine (DA) and serotonin (5-HT) neurons apparently unaffected. Administration of DSP4 to adult rats produced regional changes in the NA levels of the CNS were similar to those observed in the mouse. There were no indications of DSP4 affecting dopamine and adrenaline neurons in rat CNS, although a minor 5-HT depleting effect was noted. DSP4 treatment resulted in an increase in beta-adrenoceptor binding in vitro to homogenates from the cerebral cortex, using [3H]dihydroalprenolol as radioligand. Treatment of newborn rats with DSP4 caused permanent NA disappearance in the cerebral cortex and spinal cord, whereas marked NA increases were found in the pons-medulla and cerebellum. Administration of DSP4 to pregnant rats (gestation day 15) led to a marked and permanent NA depletion in the cerebral cortex and spinal cord in the offspring. The results support the view that DSP4 can produce an acute and relatively selective degeneration of NA nerve terminals in the rat and mouse. The results furthermore indicate that DSP4 (systemically administered) causes a preferential degeneration of NA nerve terminal projections originating from the locus coeruleus in the CNS. Since DSP4 can pass both the blood-brain and blood-placenta barrier and appears to have potent neurotoxic actions on NA neurons, DSP4 may serve as a useful denervation tool for the analysis of NA transmitter functions, particularly in the CNS of both adult and developing animals.     

Effects of antidepressant drugs,selective noradrenaline-or 5-hydroxytryptamine uptake inhibitors,on apomorphine-induced hypothermia in mice

Antidepressant drugs which are selective noradrenaline (NA) uptake inhibitors (desipramine, maprotiline, oxaprotiline, talsupram: 0.625–10 mg/kg) antagonized dose-dependently hypothermia induced by 16 mg/kg apomorphine (APO) in mice. Of the two stereoisomers of oxaprotiline, only that inhibiting NA uptake was active. Antidepressants which are selective 5-hydroxytryptamine (5-HT) uptake inhibitors (citalopram, fluvoxamine: 2.5–40 mg/kg) did not affect APO (16 mg/kg)-induced hypothermia. Neither NA nor 5-HT uptake inhibitors counteracted hypothermia induced by 1 mg/kg APO, a dose which is easily antagonized by low doses of dopamine receptor blockers. The antagonistic action of desipramine towards APO (16 mg/kg)-induced hypothermia was prevented by phenoxybenzamine, prazosin and (-)-propranolol, while (+)-propranolol and cyproheptidine were inactive. St 587 (an alpha₁-adrenoceptor agonist) or salbutamol (an agonist of beta-adrenoceptors) attenuated APO (16 mg/kg)-induced hypothermia: given jointly, the drugs completely reversed it.m-CPP, a 5-HT receptor agonist, did not affect APO (16 mg/kg)-induced hypothermia. In conclusion, the antagonistic action of antidepressant drugs towards APO (16 mg/kg)-induced hypothermia in mice did not reflect their antidepressant properties, dopamine antagonism or their action on 5-HT receptors, only their effects on the NA uptake and/or NA transmission. Both alpha₁ and beta-adrenoceptors are involved in this antagonistic action.The results were presented at the IUPHAR 9th International Congress of Pharmacology, London, 29 July–3 August, 1984 Former name: Halina Kwiatek     

Anxiolytic-like effect of milnacipran in the four-plate test in mice: mechanism of action

Milnacipran is a serotonin/noradrenaline reuptake inhibitor (SNRI) which has not yet been systematically studied preclinically or clinically for the treatment of anxiety disorders. In the four-plate test (FPT) which is known to predict anxiolytic-like activity in mice, milnacipran (4, 8, 16 and 32 mg/kg) demonstrated strong anti-punishment effects following acute administration. The anxiolytic-like effect of milnacipran was not reversed by the selective GABA(A) receptor antagonist, flumazenil (2 and 4 mg/kg), the selective alpha1-adrenoceptor antagonist, prazosin (0.5 and 2 mg/kg), the selective alpha2-adrenoceptor antagonist, idazoxan (1 and 4 mg/kg) or the selective 5-HT2B receptor antagonist, SB 206553 (0.1 and 1 mg/kg). In contrast, the selective 5-HT2A receptor antagonist, SR 46349B (0.1 and 1 mg/kg), and the non-selective 5-HT2 receptor antagonist, ketanserin (0.125 and 0.5 mg/kg), completely abolished the anxiolytic-like effect of milnacipran in FPT. Neurochemical depletion of NA or 5-HT completely abolished the activity of milnacipran. These results strongly suggest that activation of 5-HT2A receptors is critically involved in the anxiolytic activity of milnacipran. On the other hand the lack of activity of milnacipran after depletion of NA or 5-HT is consistent with milnacipran acting on the locus coeruleus to induce 5-HT release. The present data suggest a strong connection between 5-HT2A receptors and NA neurotransmission.     

Interaction studies between three antidepressant drugs (chlorimipramine, imipramine and zimelidine) and noradrenaline, tyramine and vagal stimulation on the heart rate and blood pressure in dogs

Anaesthetized Beagle dogs were given increasing intravenous doses of imipramine, chlorimipramine or zimelidine. At each dose interval the interference of the drug administered with the effects on blood pressure and heart rate of vagal stimulation, NA injection and tyramine injection was investigated. Also, the in vitro uptake of 5-HT into platelets after in vivo administration to unanaesthetized dogs of 5 mg chlorimipramine or 5 mg zimelidine was studied. Chlorimipramine and zimelidine were found to be about equipotent as regards 5 HT-uptake into platelets after in vivo administration. Imipramine and chlorimipramine potentiated the effects of NA after the 2 mg/kg dose. Imipramine but not chlorimipramine interfered with the effects of tyramine after the 4 mg/kg dose. Zimelidine did not interfere with either NA or tyramine at any dose level studied (maximal cumulative dose 62 mg/kg). The effect of vagal stimulation was significantly inhibited after 8 mg/kg (cumulative dose 14 mg/kg) of imipramine and 16 mg/kg (cumulative dose 30 mg/kg) of chlorimipramine and zimelidine, respectively. It is concluded that zimelidine in comparison with imipramine and chlorimipramine has no or at most a slight effect on peripheral adrenergic neurones. It has less pronounced anticholinergic properties than imipramine but is about equipotent to chlorimipramine in this respect.     

Effect of DSP4 on hippocampal kindling in rats

The effect of DSP4, a noradrenergic neurotoxin, on hippocampal kindling were examined in rats. The depletion of NA induced by intraperitoneal injection of DSP4 (50 mg/kg) facilitated the rate of hippocampal kindling by reducing the time spent in stage 2. The number of wet-dog shakes was also reduced in experimental animals during kindling stages 1 and 2 compared to control group. The results indicate that in DSP4 treated rats there is a more rapid activation of structures involved in the propagation of the local limbic seizure to motor centers responsible for the behavioral manifestation of the seizure, which is due to the NA depletion.     

Cyclic AMP and cyclic GMP in the rat gastric mucosa

Summary Mice were given various 7-day treatments designed to reduce cerebral noradrenergic function. Following these treatments, alterations in the sensitivity of limbic forebrain adenylate cyclase to noradrenaline (NA), and in the locomotor response of the mice to dopaminergic and noradrenergic agonists were studied. An enhanced response of the NA-sensitive adenylate cyclase to 1-NA (10^–5 M), in comparison to control mice, was demonstrated after treatments producing a chronic reduction in stimulation of -adrenergic receptors (phenoxybenzamine 10 mg/kg per day), -adrenergic receptors (propranolol, 0.05% diet), all noradrenergic receptors (reserpine 5 mg/kg, followed by FLA-63, 25 mg/kg per day), and dopaminergic as well as noradrenergic receptors (chlorpromazine, 5 mg/kg per day). Sensitivity of the NA-stimulated adenylate cyclase was not altered by chronic treatment with FLA-63 alone as a 0.05% diet. However, none of the chronic treatments except chlorpromazine administration increased the locomotor activity recorded for 2 h following apomorphine (1 mg/kg, s.c.) and none of the treatments altered the locomotor activity recorded in the 2 h following administration of the -adrenergic agonist clonidine (1 mg/kg, i.p.), or a combination of apomorphine (1 mg/kg, s.c.) plus clonidine (1 mg/kg, i.p.) The results support the hypothesis that noradrenergic stimulation plays a role secondary to that of dopamine in the production of locomotor activity.     

Hepatoprotective and anti-inflammatory activities of Plantago major L

Objective:

The aim of this study was to investigate anti-inflammatory and hepatoprotective activities of Plantago major L. (PM).

Materials and Methods:

Anti-inflammatory activity: Control and reference groups were administered isotonic saline solution (ISS) and indomethacin, respectively. Plantago major groups were injected PM in doses of 5 mg/kg (PM-I), 10 mg/kg (PM-II), 20 mg/kg (PM-III) and 25 mg/kg (PM-IV). Before and three hours after the injections, the volume of right hind-paw of rats was measured using a plethysmometer.

Hepatoprotective Activity:

The hepatotoxicity was induced by carbon tetrachloride (CCl4) administration. Control, CCl4 and reference groups received isotonic saline solution, CCl4 and silibinin, respectively. Plantago major groups received CCl4 (0.8 ml/kg) and PM in doses of 10, 20 and 25 mg/kg, respectively for seven days. Blood samples and liver were collected on the 8th day after the animals were killed.

Results:

Plantago major had an anti-inflammatory effect matching to that of control group at doses of 20 and 25 mg/kg. It was found that reduction in the inflammation was 90.01% with indomethacin, 3.10% with PM-I, 41.56% with PM-II, 45.87% with PM-III and 49.76% with PM-IV. Median effective dose (ED50) value of PM was found to be 7.507 mg/kg. Plantago major (25 mg/kg) significantly reduced the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels when compared to the CCl4 group. The histopathological findings showed a significant difference between the PM (25 mg/kg) and CCl4 groups.

Conclusion:

The results showed that PM had a considerable anti-inflammatory and hepatoprotective activities.     

Possible mechanism of the hypotensive action of 2,6-dichlorobenzylidene aminoguanidine: evidence for central noradrenaline receptor stimulation

The effects of 2,6-dichlorobenzylidene aminoguanidine acetate (DCBAG, Wy 8678, Wyeth Laboratories) have been studied on central monoamine neurons of rats in chemical and physiological experiments. Catecholamine (CA) and 5-hydroxytryptamine (5-HT) turnover was studied by the use of the tyrosine hydroxylase inhibitor, α-methyltyrosine methylester (H 44/68) and the tryptophane hydroxylase inhibitor, α-propyldopacetamide (H 22/54). The degree of CA and 5-HT depletion was studied biochemically and histochemically. Arterial pressure, heart and respiration rate were studied in rats anaesthetized with halothane, and the flexor hindlimb reflex was tested in acutely spinalized rats.Biochemically and histochemically, a dose dependent reduction of H 44/68-induced noradrenaline (NA) depletion was found both in the cerebral cortex and in the hypothalamus suggesting a reduction of NA turnover by DCBAG in doses of 0.1–10 mg/kg. The H 44/68-induced disappearance of dopamine (DA) and the H 22/54-induced disappearance of 5-HT was not changed by DCBAG, 1 mg/kg, but a significant reduction was found after 10 mg/kg. These results suggest a decrease of DA and 5-HT turnover with higher doses. The flexor hindlimb reflex, which is dependent on NA receptor activity, was increased by DCBAG (1–20 mg/kg) in spite of previous reserpine and H 44/68 treatment. It is therefore suggested that the decrease in NA turnover observed could be due to a NA receptor stimulating effect of DCBAG, eliciting a feedback to reduce NA release and turnover.DCBAG (5–500 μg/g) produced a dose-dependent reduction in arterial pressure and heart rate; higher doses also reduced respiratory rate. The CA receptor blocking agent chlorpromazine (5 mg/kg), but not the DA receptor blocking agent spiroperidol (1 mg/kg), blocked the effects of DCBAG on arterial pressure and heart rate. These findings suggest that DCBAG lowers arterial pressure and heart rate by its NA receptor stimulating action. These results give further evidence for the existence of a central NA vasodepressor mechanism.     

Influence of serotonin 5-HT(7) receptor blockade on the behavioral and neurochemical effects of imipramine in rats

The aim of the present study was to examine the effect of the selective 5-HT(7) receptor antagonist (2R)-1-[(3-hydroxyphenyl)sulfonyl]-2-[2-(4-methyl-1-piperidinyl)ethyl]pyrrolidine (SB-269970), administered alone or in combination with imipramine, on the immobility time of rats in the forced swim test as well as on the extracellular levels of dopamine (DA), noradrenaline (NA), serotonin (5-HT) and their metabolites in the prefrontal cortex of freely moving rats. Both compounds were administered intraperitoneally (ip). Like imipramine (30 mg/kg, but not 20 mg/kg), SB-269970 (1.25 and 2.5 mg/kg, but not 0.625 mg/kg) significantly shortened the immobility time of rats without affecting their exploratory locomotor activity measured in the open field test. SB-269970 (0.625 and 1.25 mg/kg) raised the extracellular levels of DA, NA, 5-HT and their metabolites in rat prefrontal cortex. In that structure, imipramine (20 mg/kg) produced an increase in all the neurotransmitters measured, but failed to affect the levels of their metabolites. A combination of the inactive doses of SB-269970 (0.625 mg/kg) and imipramine (20 mg/kg) found in the forced swim test produced antidepressant-like effect, which did not stem from the increased exploratory locomotor activity. At the same time, that combination voked a vast increase in the output of NA - but not DA and 5-HT - compared to the effects of both those drugs given alone. These results open up a possibility that the stimulating effect of SB-269970 on DA, NA and 5-HT transmission in the prefrontal cortex plays some role in the antidepressant-like activity of this compound. Moreover, these findings suggest that the increase in cortical NA level seems to account for the anti-immobility action observed after joint administration of the selective 5-HT(7) receptor antagonist and imipramine in rats.     

In vivo, noradrenaline is a substrate for rat brain monoamine oxidase A and B

In vivo clorgyline (5 mg/kg) and (-)-deprenyl (5 mg/kg) selectively inhibit monoamine oxidase (MAO) type A and B activities in rat brain hypothalamus and caudate nucleus using 5-hydroxytryptamine (5-HT), noradrenaline (NA), and beta-phenylethylamine (PEA) as substrates. Clorgyline induces a significant increase in NA concentrations of hypothalamus and caudate nucleus; however (-)-deprenyl is without effect. The combination of clorgyline and (-)-deprenyl at the above doses completely inhibits both forms of MAO, resulting in an even greater increase in NA levels in both brain areas than observed with clorgyline. The non-selective inhibitor tranylcypromine (5 mg/kg) produced a similar effect. Rats pretreated with the selective or the non-selective inhibitors but given L-DOPA (50 mg/kg) have a similar pattern of brain NA, but its concentrations are higher in both brain regions. The results indicate that although in vitro NA may be an exclusive substrate for MAO type A, in vivo, when this enzyme form is selectively inhibited, NA at high concentrations can be a substrate for MAO type B.     

Effects of acute and chronic administration of MCI-225, a new selective noradrenaline reuptake inhibitor with 5-HT3 receptor blocking action, on extracellular noradrenaline levels in the hypothalamus of stressed rats

In the present study, we investigated the effects of acute and chronic systemic administration of MCI-225 (4-(2-fluorophenyl)-6-methyl-2-(1-piperazinyl)thieno[2,3-d]pyrimidine monohydrate hydrochloride), a newly-developed selective noradrenaline (NA) reuptake inhibitor with 5-HT3-receptor-blocking action, on extracellular NA levels in the hypothalamus of stressed and non-stressed rats by utilizing intracerebral microdialysis. Acute administration of MCI-225 (3 and 10 mg/kg, p.o.) significantly and dose-dependently increased extracellular NA levels in the hypothalamus in non-stressed rats. Footshock for 20 min also significantly increased NA levels in the hypothalamus of both groups of rats pretreated with vehicle and MCI-225. Although chronic administration of MCI-225 (3 or 10 mg/kg, p.o. for 14 days) did not alter the basal extracellular NA levels in the hypothalamus, the stress-induced increases in extracellular NA levels were significantly lower in rats chronically treated with MCI-225 (10 mg/kg) than those of rats pretreated with vehicle for the same period. The increase in extracellular NA levels induced by MCI-225 challenge (3 or 10 mg/kg, p.o.) were not different between rats chronically treated with MCI-225 or vehicle. These results suggest that MCI-225 enhances extracellular NA levels in the hypothalamus in both non-stressed and stressed rats by inhibiting NA uptake and that chronic systemic administration of MCI-225 did not alter basal extracellular NA levels, but reduced the increase in NA release caused by footshock stress. These data suggest the possibility that MCI-225 might possess anxiolytic and/or antidepressant properties.     

The dopamine autoreceptor agonist, B-HT 920, preferentially reduces brain dopamine release in vivo: biochemical indices of brain dopamine, noradrenaline and serotonin in ventriculocisternal perfusates in the cat

B-HT 920 (6-allyl-2-amino-5,6,7,8-tetrahydro-4H-thiazolo[4,5-d]azepine), a candidate for selective dopamine (DA) autoreceptor agonist activity, was tested for its interactions with biochemical parameters of brain dopaminergic, noradrenergic and serotoninergic systems as measured in ventriculocisternal perfusates of chloralose-anaesthetized cats. DA, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), noradrenaline (NA) and 5-hydroxyindolic acid (5-HIAA) were measured in samples of 30 min collection periods by high-pressure liquid chromatography with electrochemical detection. B-HT 920, in the dose range of 0.03-1 mg/kg i.v., promptly inhibited the efflux of DA and DOPAC in a dose-dependent manner. The 1 mg/kg dose of B-HT 920 reduced the DA levels below 25% of control levels for the whole length of the experiments. The HVA levels were reduced less and in a protracted manner. Only the highest dose of B-HT 920 tested (1 mg/kg) had a significant effect on the level of NA (marked, prompt reduction) and 5-HIAA (delayed, moderate reduction), reflecting its well known alpha 2-adrenoceptor agonist property. The effects of B-HT 920 on the dopaminergic indices were DA receptor-mediated as they were reversed by a low dose (0.05 mg/kg i.v.) of haloperidol. In contrast, the alpha 2-adrenoceptor blocking drug, idazoxan, 4 mg/kg i.v., while it reversed the NA and 5-HIAA reductions did not modify the effect of B-HT 920 on DA, DOPAC and HVA. Thus B-HT 920, in the dose range between 0.03-0.1 mg/kg, selectively affected brain dopaminergic parameters. Our experiments demonstrated that B-HT 920 causes an effective, long lasting and selective suppression of extracellular brain DA levels in vivo. B-HT 920 represents a promising compound for clinical use in pathological conditions known to be ameliorated by a reduction of brain DA activity, such as Huntington's disease, mania and schizophrenia.     

Idazoxan and 8-OH-DPAT modify the behavioral effects induced by either NA, or 5-HT, or dual NA/5-HT reuptake inhibition in the rat forced swimming test.

The rat forced swimming test (FST) predicts the efficacy of antidepressants, which decrease immobility duration in the test, and can distinguish selective serotonin (5-HT) and noradrenaline (NA) reuptake inhibitors, which, respectively, increase swimming and climbing behaviors. However, dual 5-HT and NA reuptake-inhibition produces climbing behavior solely, thereby suggesting with other data that the NA-system mediates inhibiting interactions on 5-HT-induced swimming in the FST. Since alpha(2)-adrenoreceptors and 5-HT(1A)-receptors have important regulatory functions and are involved in 5-HT/NA interactions, we examined whether the alpha(2)-receptor-antagonist idazoxan and the 5-HT(1A)-receptor-agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) would modify the behavioral pattern induced in the FST by either selective or non-selective antidepressant treatments. The rats were treated subacutely (3 injections IP over 48 h) with: (a) idazoxan (0.5-10 mg/kg) alone, and in combination with desipramine (10 mg/kg), or desipramine + fluoxetine (10/10 mg/kg), or the dual serotonin/noradrenaline reuptake-inhibitor milnacipran (20 mg/kg). (b) 8-OH-DPAT (0.25-1 mg/kg) alone, and in combination with either desipramine (10 mg/kg) or fluoxetine (10 mg/kg). The results indicated: (a) Idazoxan (0.5, 5, 10 mg/kg) produced no anti-immobility effects per se in the FST, antagonized the effects of the NA-reuptake-inhibitor desipramine, and allowed desipramine + fluoxetine, as well as milnacipran, to increase swimming behavior. (b) 8-OH-DPAT produced non-significant effects per se, potentiated desipramine-induced antidepressant-like effects on immobility and climbing, and both antagonized swimming and produced climbing behavior in combination with fluoxetine. Our data support clinical trials suggesting that alpha(2)-receptor-antagonists and 5-HT(1A)-receptor-agonists may be of interest in augmentation strategies for antidepressant treatments. The scoring of active behaviors in the FST appears to be an interesting tool for studying 5-HT/NA interactions induced by antidepressants, as well as for the testing of augmentation strategies.