Nasser Haddjeri

GABAergic Network Activation of Glial Cells Underlies Hippocampal Heterosynaptic Depression

Tetanus-induced heterosynaptic depression in the hippocampus is a key cellular mechanism in neural networks implicated in learning and memory. A growing body of evidence indicates that glial cells are important modulators of synaptic functions, but very little is known about their role in heterosynaptic plasticity. We examined the role of glial cells in heterosynaptic depression, knowing that tetanization and NMDA application caused depression of synaptic field responses (fEPSPs) and induced Ca2+ rise in glial cells. Here we report that chelating Ca2+ in a glial syncytium interfered with heterosynaptic depression and NMDA-induced fEPSP depression, suggesting that Ca2+ activation of glial cells is necessary for heterosynaptic depression. The NMDA-induced Ca2+ rise in glial cells was sensitive to tetrodotoxin and reduced by the GABAB antagonist CGP55845. Both heterosynaptic depression and simultaneous Ca2+ activation of glial cells were prevented by CGP55845, suggesting an involvement of the GABAergic network in glial activation and heterosynaptic depression. Also, the GABAB agonist baclofen caused both a Ca2+ rise in glial cells and fEPSP depression. Heterosynaptic depression, as well as NMDA- and baclofen-induced depression, were attenuated by an A1 antagonist, cyclopentyl-theophylline, whereas glial cell activation was not, indicating a role of adenosine downstream of glial activation. Finally, heterosynaptic depression requires ATP degradation because ectonucleotidase inhibitors reduced this plasticity. Our work indicates that Ca2+ activation of glial cells is necessary for heterosynaptic depression, which involves the sequential interaction of Schaffer collaterals, the GABAergic network, and glia. Thus, glial and neuronal networks are functionally associated during the genesis of heterosynaptic plasticity at mammalian central excitatory synapses.

Modulation of the firing activity of noradrenergic neurones in the rat locus coeruleus by the 5-hydroxtryptamine system

The aim of the present study was to investigate the putative modulation of locus coeruleus (LC) noradrenergic (NA) neurones by the 5‐hydroxytryptaminergic (5‐HT) system by use of in vivo extracellular unitary recordings and microiontophoresis in anaesthetized rats. To this end, the potent and selective 5‐HT1A receptor antagonist WAY 100635 (N‐{2‐[4(2‐methoxyphenyl)‐1‐piperazinyl]ethyl}‐N‐(2‐pyridinyl)cyclohexanecarboxamide trihydroxychloride) was used. In the dorsal hippocampus, both local (by microiontophoresis, 20 nA) and systemic (100 μg kg−1, i.v.) administration of WAY 100635 antagonized the suppressant effect of microiontophorectically‐applied 5‐HT on the firing activity of CA3 pyramidal neurones, indicating its antagonistic effect on postsynaptic 5‐HT1A receptors. WAY 100635 and 5‐HT failed to modify the spontaneous firing activity of LC NA neurones when applied by microiontophoresis. However, the intravenous injection of WAY 100635 (100 μg kg−1) readily suppressed the spontaneous firing activity of LC NA neurones. The lesion of 5‐HT neurones with the neurotoxin 5,7‐dihydroxytryptamine increased the spontaneous firing activity of LC NA neurones and abolished the suppressant effect of WAY 100635 on the firing activity of LC NA neurones. In order to determine the nature of the 5‐HT receptor subtypes mediating the suppressant effect of WAY 100635 on NA neurone firing activity, several 5‐HT receptor antagonists were used. The selective 5‐HT3 receptor antagonist BRL 46470A (10 and 100 μg kg−1, i.v.), the 5‐HT1D receptor antagonist GR 127935 (100 μg kg−1, i.v.) and the 5‐HT1A/1B receptor antagonist (−)‐pindolol (15 mg kg−1, i.p.) did not prevent the suppressant effect of WAY 100635 on the firing activity of LC NA neurones. However, the suppressant effect of WAY 100635 was prevented by the non‐selective 5‐HT receptor antagonists spiperone (1 mg kg−1, i.v.) and metergoline (1 mg kg−1, i.v.), by the 5‐HT2 receptor antagonist ritanserin (500 μg kg−1, i.v.). It was also prevented by the 5‐HT1A receptor/α1D‐adrenoceptor antagonist BMY 7378 (1 mg kg−1, i.v.) and by the α1‐adrenoceptor antagonist prazosin (100 μg kg−1, i.v.). These data support the notion that the 5‐HT system tonically modulates NA neurotransmission since the lesion of 5‐HT neurones enhanced the LC NA neurones firing activity and the suppressant effect of WAY 100635 on the firing activity of NA neurones was abolished by this lesion. However, the location of the 5‐HT1A receptors involved in this complex circuitry remains to be elucidated. It is concluded that the suppressant effect of WAY 100635 on the firing activity of LC NA neurones is due to an enhancement of the function of 5‐HT neurones via a presynaptic 5‐HT1A receptor. In contrast, the postsynaptic 5‐HT receptor mediating this effect of WAY 100635 on NA neurones appears to be of the 5‐HT2A subtype.

Effects of the co-administration of mirtazapine and paroxetine on serotonergic neurotransmission in the rat brain

The alpha(2)-adrenoreceptor antagonist mirtazapine, which is also a 5-HT(2), 5-HT(3) and H(1) receptors antagonist and the selective serotonin (5-HT) reuptake inhibitor paroxetine are effective antidepressant drugs which enhance 5-HT neurotransmission via different mechanisms. The present studies were undertaken to determine whether the mirtazapine-paroxetine combination could induce an earlier and/or a greater effect on the 5-HT system than either drug alone. Using in vivo electrophysiological paradigms, the firing activity of dorsal raphe 5-HT neurons was decreased by 70% in rats treated with paroxetine (10 mg/kg/day, s.c.) for 2 days and was back to normal after 21 days. In contrast, a 2-day treatment with mirtazapine (5 mg/kg/day, s.c.) did not alter the firing of 5-HT neurons whereas it was increased by 60% after 21 days of treatment. A low dose of mirtazapine (5 mg/kg/day, s.c.x2 days) failed to offset the decremental effect of paroxetine on the 5-HT neuron firing activity, but a higher dose (10 mg/kg/day, s.c.x2 days) did attenuate the decremental effect of paroxetine. In the dorsal hippocampus, neither mirtazapine (5 mg/kg/day, s.c.) nor a paroxetine (10 mg/kg/day, s.c.) treatment altered the responsiveness of 5-HT(1A) receptors to microiontophoretically-applied 5-HT. Both in controls and in rats treated for 2 days with paroxetine alone, the administration of the 5-HT(1A) antagonist WAY 100635 (25-100 microg/kg, i.v.) did not change the firing activity of dorsal hippocampus CA(3) pyramidal neurons. However, WAY 100635 increased significantly the firing activity of these neurons in rats treated with mirtazapine alone but to a greater extent with both mirtazapine and paroxetine for 2 days. After 21 days of treatment, WAY 100635 increased to a greater degree the firing rate of CA(3) pyramidal neurons in rats which received the combination over rats given either drug alone. It is concluded that the mirtazapine-paroxetine combination shortened the delay in enhancing the tonic activation of postsynaptic 5-HT(1A) receptors and produced a greater activation of the postsynaptic 5-HT(1A) receptors than either drug given alone. The present results suggested that mirtazapine may have a faster onset of action than a SSRI, and that the co-administration of mirtazapine and paroxetine may accelerate the antidepressant response and as well as being more effective than either drug alone.

Effects of long-term treatment with the alpha 2-adrenoceptor antagonist mirtazapine on 5-HT neurotransmission.

Abstract Mirtazapine (ORG 3770, Remeron®) is a non-selective α2-adrenoceptor antagonist with antidepressant activity in major depression. The aim of the present study was to assess, using an in vivo electrophysiological paradigm, the effect of long-term treatment with mirtazapine on pre- and postsynaptic α2-adrenoceptors and on 5-HT neurotransmission in male Sprague-Dawley rats. A 21-day treatment with mirtazapine (5u2009mg/kg/day, s.c., using osmotic minipumps) increased the spontaneous firing activity of locus coeruleus noradrenaline (NA) neurons. Their firing activity was back to normal 48u2009h after removing the minipump. However, this treatment did not modify the dose-response curve of the suppressant effect of the α2-adrenoceptor agonist clonidine on the firing activity of NA neurons. The spontaneous firing activity of dorsal raphe 5-HT neurons was also markedly increased in mirtazapine-treated rats, and was back to normal 48u2009h after removing of the minipump. The dose-response curve of the suppressant effect of clonidine on the firing activity of 5-HT neurons was altered in mirtazapine-treated rats. Furthermore, it was further shifted to the left after a 48-h washout. Long-term mirtazapine treatment did not modify the suppressant effects of microiontophoretically-applied NA and 5-HT on the firing activity of CA3 dorsal hippocampus pyramidal neurons. However, this mirtazapine treatment antagonized both the enhancing effect of a low dose (10u2009μg/kg, i.v.) and the reducing effect of a high dose (100u2009μg/kg, i.v.) of the α2-adrenoceptor agonist clonidine on the effectiveness of the electrical stimulation of the ascending 5-HT pathway in suppressing the firing activity of dorsal hippocampus CA3 pyramidal neurons. After a 48-h washout, only the effect of the high dose of clonidine was attenuated, suggesting a desensitization of the terminal α2-adrenergic heteroreceptor, but not of the terminal α2-adrenergic autoreceptor. The decrease in the effectiveness of the stimulation upon increasing its frequency from 1 to 5u2009Hz (due to the activation of terminal 5-HT autoreceptors) was unaltered after the long-term mirtazapine treatment. In conclusion, the tonic activation of postsynaptic 5-HT receptors is enhanced by a 21-day treatment with mirtazapine, as a result of a sustained increase in 5-HT neuron firing activity in the presence of decreased function of α2-adrenergic heteroreceptors located on 5-HT terminals in the dorsal hippocampus.

Acute and long-term actions of the antidepressant drug mirtazapine on central 5-HT neurotransmission.

Mirtazapine (ORG 3770, Remeron) is a new alpha 2-adrenoceptor antagonist which has been shown to be an effective antidepressant drug. The aims of the studies were to assess, using an in vivo electrophysiological paradigm in the rat, the effects of acute and long-term treatment with mirtazapine on pre- and postsynaptic alpha 2-adrenoceptors and to determine whether this drug could modulate serotonin (5-HT) neurotransmission. Acute administration of mirtazapine produced a transient increase of the firing activity of dorsal raphe 5-HT neurons. This effect was mediated via norepinephrine (NE) neurons because it was abolished in NE-lesioned rats. In fact, this increased firing rate of 5-HT neurons was due to their activation by the enhanced release of NE resulting from the blockade of alpha 2-adrenergic autoreceptors of locus coeruleus neurons. Furthermore, acute mirtazapine injection transiently enhanced the firing activity of locus coeruleus NE neurons and attenuated the suppressant effect of the alpha 2-adrenoceptor agonist clonidine on these NE neurons. Sustained administration of mirtazapine for 21 days (5 mg/kg/day, s.c., using minipumps) lead to a marked increase in the firing rate of 5-HT neurons (75%) but a more modest increase in the firing rate of NE neurons (30%), as well as to a desensitization of alpha 2-adrenergic heteroreceptors on 5-HT terminals in the hippocampus. The desensitization of these heteroreceptors, resulting from an increased synaptic availability of NE induced by mirtazapine would free 5-HT terminals from the inhibitory influence of NE on 5-HT release. These modifications of 5-HT neurons lead to an increased tonic activation of postsynaptic 5-HT1A receptors. The latter conclusion was based on the capacity of the selective 5-HT1A receptor antagonist WAY 100635 to enhance the firing activity of dorsal hippocampus CA3 pyramidal neurons in mirtazapine-treated rats but not in controls. This enhanced 5-HT neurotransmission may underlie to the antidepressant effect of mirtazapine.

Increased Tonic Activation of Rat Forebrain 5-HT1A Receptors by Lithium Addition to Antidepressant Treatments

The present study was undertaken to determine whether lithium addition to long-term treatment with different classes of antidepressant drugs could induce a greater effect on the serotonin (5-HT) system than the drugs given alone. Because 5-HT1A receptor activation hyperpolarizes and inhibits the firing activity of CA3 pyramidal neurons in the dorsal hippocampus, the degree of disinhibition produced by the selective 5-HT1A receptor antagonist WAY 100635 was determined using in vivo extracellular recordings. In controls, as well as in rats receiving a lithium diet for 3 days, the administration of WAY 100635 (25-100 μg/kg, IV) did not modify the firing activity of dorsal hippocampus CA3 pyramidal neurons. When the tricyclic antidepressant imipramine (10 mg/kg/day, SC), the monoamine oxidase inhibitor tranylcypromine (2.5 mg/kg/day, SC) and the selective 5-HT reuptake inhibitor paroxetine (10 mg/kg/day, SC) were administered alone for 21 days, a dose of 50 μg/kg of WAY 100635 was needed to increase significantly the firing activity of these neurons. On the other hand, WAY 100635, at a dose of only 25 μg/kg, increased significantly the firing rate of CA3 pyramidal neurons in rats receiving both a long-term antidepressant treatment and a short-term lithium diet. It is concluded that the addition of lithium to antidepressant treatments produced a greater disinhibition of dorsal hippocampus CA3 pyramidal neurons than any treatments given alone. The present results support the notion that the addition of lithium to antidepressants may produce a therapeutic response in treatment-resistant depression by enhancing 5-HT neurotransmission.

Effect of sustained administration of the 5-HT1A receptor agonist flesinoxan on rat 5-HT neurotransmission

A short-term treatment with flesinoxan (2.5 and 5 mg/kg/day x 2 days, s.c., delivered using osmotic minipumps) decreased significantly the spontaneous firing activity of dorsal raphe serotonin (5-HT) neurons of male Sprague-Dawley rats. This firing was still decreased following 1 week of treatment with flesinoxan (5 mg/kg/day) but was back to normal after a treatment of 2 weeks. This recovery of firing was associated with a 3-fold shift to the right of the dose-response curve of the effect of the 5-HT autoreceptor agonist lysergic acid diethylamide on the firing activity of 5-HT neurons, indicating a desensitization of somatodendritic 5-HT1A autoreceptors. At the postsynaptic level, long-term treatment with flesinoxan (5 mg/kg/day x 14 days) did not modify the responsiveness of dorsal hippocampus CA3 pyramidal neurons to microiontophoretic applications of 5-HT and flesinoxan nor to endogenous 5-HT released by the electrical stimulation of the ascending 5-HT pathway, indicating an unchanged sensitivity of postsynaptic 5-HT1A receptors. Finally, in rats treated with flesinoxan for 2 weeks, the administration of the selective 5-HT1A receptor antagonist (N-{2-[4(2-methoxyphenyl)-1-piperazinyl]ethyl}-N-(2-pyridinyl)cyclohe xanecarboxamide trihydroxychloride (WAY 100635, 100 and 500 microg/kg, i.v.) did not increase the firing activity of dorsal hippocampus CA3 pyramidal neurons, thus failing to reveal an enhanced tonic activation of postsynaptic 5-HT1A receptors as for other antidepressant drugs, including the 5-HT1A receptor agonist gepirone. The marked potency and the long dissociation constant of flesinoxan for the 5-HT1A receptors may account for the latter discrepancy. In conclusion, as for selective 5-HT re-uptake inhibitors, monoamine oxidase inhibitors and 5-HT1A receptor agonists, flesinoxan produced most of the adaptive changes exerted by these antidepressant drugs on the 5-HT system.

Modulation of the firing activity of rat serotonin and noradrenaline neurons by (±)pindolol

BACKGROUNDn(+/-)Pindolol is a beta-adrenergic/5-HT1A receptor antagonist used in combination with certain antidepressant drugs to accelerate the onset of the antidepressive response.nnnMETHODSnThe aim of the present study was to assess, using an in vivo electrophysiologic paradigm, the effect of (+/-)pindolol on the spontaneous firing activity of rat dorsal raphe serotonin (5-HT) and locus coeruleus noradrenaline (NA) neurons.nnnRESULTSn(+/-)Pindolol did not modify the firing activity of dorsal raphe 5-HT neurons at low doses (10 and 200 micrograms/kg, i.v.), but it prevented the suppressant effect of the 5-HT autoreceptor agonist lysergic acid diethylamide (LSD, 10 micrograms/kg, i.v.) but not that of the 5-HT1A receptor 8-hydroxy-N,N-dipropyl-aminotetralin (8-OHDPAT, 5 micrograms/kg, i.v.). At a higher dose (500 micrograms/kg, i.v.), (+/-)pindolol decreased 5-HT neuronal firing and this effect was reversed by the selective 5-HT1A receptor antagonist WAY 100635 (100 micrograms/kg, i.v.), suggesting that it could act as a partial 5-HT1A autoreceptor agonist. In the locus coeruleus, the high dose of (+/-)pindolol decreased the firing activity of NA neurons and this effect was reversed by the 5-HT2A receptor antagonist MDL 100907 (200 micrograms/kg, i.v.). Finally, both a lesion of NA neurons and the administration of MDL 100907 prevented the suppressant effect of (+/-)pindolol on the firing of 5-HT neurons.nnnCONCLUSIONSnIt is suggested that, at low doses, (+/-)pindolol acts as a somatodendritic 5-HT1A autoreceptor antagonist whereas at a higher dose, it decreases the tonic excitatory input from NA neurons to 5-HT neurons.

The Effects of Mirtazapine on the Interactions between Central Noradrenergic and Serotonergic Systems

SummarySeveral lines of evidence have established that antidepressant drug treatments alter the serotonin (5-hydroxytryptamine; 5-HT) and noradrenaline (norepinephrine) systems. However, very few studies on the interactions between these two systems have been carried out. The characterisation of these interactions seems to be crucial to a better understanding of the mechanism of action of antidepressant drugs.We have studied the effects of the novel antidepressant mirtazapine (Remeron®, Org 3770) on the interaction between central noradrenergic and serotonergic systems using an in vivo electrophysiological paradigm in the rat. Our results suggest that (±)mirtazapine is an antagonist of both α2-adrenergic auto- and heteroreceptors, whereas (−)mirtazapine is selective for the α2-heteroreceptor and, according to the dose used, can activate terminal serotonin autoreceptors. In view of the important role of α2-adrenoceptors in controlling serotonergic neurotransmission, selective α2-adrenoceptor antagonists with indirect serotonin enhancing effects may represent a new generation of antidepressant drugs.

Effect of the reversible monoamine oxidase-A inhibitor befloxatone on the rat 5-hydroxytryptamine neurotransmission.

The aim of the present study was to assess, using in vivo electrophysiological paradigms, the effect of sustained administration of the selective and reversible monoamine oxidase-A inhibitor beflotaxone on serotonin (5-hydroxytryptamine, 5-HT) neurotransmission. In male Sprague-Dawley rats with the osmotic minipumps in place, a treatment with befloxatone (0.75 mg/kg per day, s.c.) for 2 days decreased the spontaneous firing activity of dorsal raphe 5-HT neurons. The combination of befloxatone and the 5-HT1A/1B receptor antagonist (-)-pindolol (15 mg/kg per day, s.c.) for 2 days slightly increased the firing activity of 5-HT neurons, whereas a treatment with (-)-pindolol alone for 2 days did not modify this parameter. The suppressant effects on the firing activity of 5-HT neurons of the 5-HT autoreceptor agonist lysergic acid diethylamide (LSD), injected intravenously, and of both 5-HT and the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), applied by microiontophoresis, were attenuated in rats treated with befloxatone for 2 days, suggesting an early desensitization of the somatodendritic 5-HT1A receptors. The firing activity of 5-HT neurons was back to normal after a treatment for 21 days with befloxatone but the suppressant effects of LSD, 5-HT or 8-OH-DPAT was the same as in controls. In contrast, the suppressant effect of the alpha2-adrenoceptor agonist clonidine on the firing activity of 5-HT neurons was significantly attenuated after the treatment with befloxatone for 21 days. At the postsynaptic level, the administration of the selective 5-HT1A receptor antagonist (N-[2-[4(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohe xanecarboxamide trihydroxychloride (WAY 100635, 100 microg/kg, i.v.) did not modify the firing activity of quisqualate-activated dorsal hippocampus CA3 pyramidal neurons in control rats. In contrast, in rats treated with befloxatone in combination with (-)-pindolol for 2 days as well as with befloxatone alone for 21 days, WAY 100635 significantly increased the firing of CA3 pyramidal neurons. In conclusion, these data suggest that when the firing activity of 5-HT neurons is normal in the presence of befloxatone, either after a two-day treatment together with (-)-pindolol or alone for 21 days, the tonic activation of postsynaptic 5-HT1A receptors is enhanced.