Spontaneous resurgence of conditioned fear weeks after successful extinction in brain injured mice.

Mild traumatic brain injury (TBI) is a major risk factor for post-traumatic stress disorder (PTSD), and both disorders share common symptoms and neurobiological defects. Relapse after successful treatment, known as long-term fear resurgence, is common in PTSD patients and a major therapeutic hurdle. We induced a mild focal TBI by controlled cortical impact (CCI) in male C57BL/6 J mice and used fear conditioning to assess PTSD-like behaviors and concomitant alterations in the fear circuitry. We found for the first time that mild TBI, and to a lesser extent sham (craniotomy), mice displayed a spontaneous resurgence of conditioned fear when tested for fear extinction memory recall, despite having effectively acquired and extinguished conditioned fear 6 weeks earlier in the same context. Other characteristic symptoms of PTSD are risk-taking behaviors and cognitive deficits. CCI mice displayed risk-taking behaviors, behavioral inflexibility and reductions in processing speed compared to naïve mice. In conjunction with these changes there were alterations in amygdala morphology 3 months post-trauma, and decreased myelin basic protein density at the primary lesion site and in distant secondary sites such as the hippocampus, thalamus, and amygdala, compared to sham mice. Furthermore, activity-dependent brain-derived neurotrophic factor (BDNF) transcripts were decreased in the prefrontal cortex, a key region for fear extinction consolidation, following fear extinction training in both TBI and, to a lesser extent, sham mice. This study shows for the first time that a mild brain injury can generate a spontaneous resurgence of conditioned fear associated with defective BDNF signalling in the prefrontal cortex, PTSD-like behaviors, and have enduring effects on the brain.

Behavioral and neurochemical characterization of TrkB-dependent mechanisms of agomelatine in glucocorticoid receptor-impaired mice.

Growing evidence indicates that impairment of the stress response, in particular the negative feedback regulation mechanism exerted by the hypothalamo-pituitary-adrenal (HPA) axis, might be responsible for the hippocampal atrophy observed in depressed patients. Antidepressants, possibly through the activation of BDNF signaling, may enhance neuroplasticity and restore normal hippocampal functions. In this context, glucocorticoid receptor-impaired (GR-i) mice-a transgenic mouse model of reduced GR-induced negative feedback regulation of the HPA axis-were used to investigate the role of BDNF/TrkB signaling in the behavioral and neurochemical effects of the new generation antidepressant drug, agomelatine. GR-i mice exhibited marked alterations in depressive-like and anxiety-like behaviors, together with a decreased cell proliferation and altered levels of neuroplastic and epigenetic markers in the hippocampus. GR-i mice and their wild-type littermates were treated for 21 days with vehicle, agomelatine (50mg/kg/day; i.p) or the TrkB inhibitor Ana-12 (0.5mg/kg/day, i.p) alone, or in combination with agomelatine. Chronic treatment with agomelatine resulted in antidepressant-like effects in GR-i mice and reversed the deficit in hippocampal cell proliferation and some of the alterations of mRNA plasticity markers in GR-i mice. Ana-12 blocked the effect of agomelatine on motor activity as well as its ability to restore a normal hippocampal cell proliferation and expression of neurotrophic factors. Altogether, our findings indicate that agomelatine requires TrkB signaling to reverse some of the molecular and behavioral alterations caused by HPA axis impairment.

Brain plasticity and cognitive functions after ethanol consumption in C57BL/6J mice.

Acute or chronic administrations of high doses of ethanol in mice are known to produce severe cognitive deficits linked to hippocampal damage. However, we recently reported that chronic and moderate ethanol intake in C57BL/6J mice induced chromatin remodeling within the Bdnf promoters, leading to both enhanced brain-derived neurotrophic factor (BDNF) expression and hippocampal neurogenesis under free-choice protocol. We performed here a series of cellular and behavioral studies to analyze the consequences of these modifications. We showed that a 3-week chronic free-choice ethanol consumption in C57BL/6J mice led to a decrease in DNA methylation of the Bdnf gene within the CA1 and CA3 subfields of the hippocampus, and upregulated hippocampal BDNF signaling pathways mediated by ERK, AKT and CREB. However, this activation did not affect long-term potentiation in the CA1. Conversely, ethanol intake impaired learning and memory capacities analyzed in the contextual fear conditioning test and the novel object recognition task. In addition, ethanol increased behavioral perseveration in the Barnes maze test but did not alter the mouse overall spatial capacities. These data suggested that in conditions of chronic and moderate ethanol intake, the chromatin remodeling leading to BDNF signaling upregulation is probably an adaptive process, engaged via epigenetic regulations, to counteract the cognitive deficits induced by ethanol.

Hippocampal and behavioral dysfunctions in a mouse model of environmental stress: normalization by agomelatine.

Stress-induced alterations in neuronal plasticity and in hippocampal functions have been suggested to be involved in the development of mood disorders. In this context, we investigated in the hippocampus the activation of intracellular signaling cascades, the expression of epigenetic markers and plasticity-related genes in a mouse model of stress-induced hyperactivity and of mixed affective disorders. We also determined whether the antidepressant drug agomelatine, a MT1/MT2 melatonergic receptor agonist/5-HT2C receptor antagonist, could prevent some neurobiological and behavioral alterations produced by stress. C57BL/6J mice, exposed for 3 weeks to daily unpredictable socio-environmental stressors of mild intensity, were treated during the whole procedure with agomelatine (50 mg kg(-1) per day, intraperitoneal). Stressed mice displayed robust increases in emotional arousal, vigilance and motor activity, together with a reward deficit and a reduction in anxiety-like behavior. Neurobiological investigations showed an increased phosphorylation of intracellular signaling proteins, including Atf1, Creb and p38, in the hippocampus of stressed mice. Decreased hippocampal level of the repressive epigenetic marks HDAC2 and H3K9me2, as well as increased level of the permissive mark H3K9/14ac suggested that chronic mild stress was associated with increased gene transcription, and clear-cut evidence was further indicated by changes in neuroplasticity-related genes, including Arc, Bcl2, Bdnf, Gdnf, Igf1 and Neurod1. Together with other findings, the present data suggest that chronic ultra-mild stress can model the hyperactivity or psychomotor agitation, as well as the mixed affective behaviors often observed during the manic state of bipolar disorder patients. Interestingly, agomelatine could normalize both the behavioral and the molecular alterations induced by stress, providing further insights into the mechanism of action of this new generation antidepressant drug.

Effects of exercise on mitochondrial function, neuroplasticity and anxio-depressive behavior of mice.

The present study was aimed at analyzing the effects of physical exercise on mitochondrial physiology, anxio-depressive-like behaviors and neuroplasticity in mice. Adult C57BL/6J male mice were isolated in home cages equipped or not with free-running wheels. After 6weeks of exercise, mice were tested in various behavioral paradigms to evaluate anxiety- and depressive-like behaviors. The hippocampi were dissected for neurochemical assays, including mitochondrial activity, monoamines content and the expression of genes involved in energy metabolism and brain-derived neurotrophic factor (BDNF) regulation. Exercise decreased anxiety-like behaviors in the open field and elevated plus maze, and exerted antidepressant-like effects in the tail suspension test. Exercise stimulated brain mitochondrial activity and increased resistance against rotenone, an inhibitor of complex I activity. Furthermore, mRNA expression of Bdnf, Gdnf, Tfam (mitochondrial transcription factor A), and Ndufa6 (mitochondrial I subunit) genes, as well as the phosphorylation of cAMP response element-binding protein were increased after exercise. In summary, exercise appears to engage mitochondrial pathways and to potentiate neuroplasticity and might be associated to mood improvement.

RNA splicing and editing modulation of 5-HT(2C) receptor function: relevance to anxiety and aggression in VGV mice.

Changes in serotonin(2C) receptor (5-HTR2c) editing, splicing and density were found in conditions such as depression and suicide, but mechanisms explaining the changes in 5-HTR2c function are unknown. Thus, mice expressing only the fully edited VGV isoform of 5-HTR2c, in which clinically relevant behavioral changes are associated with alterations in splicing and receptor density, were studied. VGV mice displayed enhanced anxiety-like behavior in response to a preferential 5-HTR2c agonist in the social interaction test. Nearly half of interactions between pairs of VGV congeners consisted of fighting behaviors, whereas no fighting occurred in wild-type (WT) mice. VGV mice also exhibited a striking increase in freezing behaviors in reaction to an innately aversive ultrasonic stimulus. This behavioral phenotype occurred in conjunction with decreased brain 5-HT turnover during stress. These functional data were put in relation with the 5-HTR2c mRNA splicing process generating a truncated protein (5-HTR2c-Tr) in addition to the full-length receptor (5-HTR2c-Fl). 5-HTR2c-Tr mRNA was less abundant in many brain regions of VGV mice, which concomitantly had more 5-HTR2c than WT mice. Fluorescence resonance energy transfer and bioluminescence resonance energy transfer studies in transfected living HEK293T cells showed that 5-HTR2c-Tr interacts with 5-HTR2c-Fl. The 5-HTR2c-Tr was localized in the endoplasmic reticulum where it retained 5-HTR2c-Fl, preventing the latter to reach the plasma membrane. Consequently, 5-HTR2c-Tr decreased (3)H-mesulergine binding to 5-HTR2c-Fl at the plasma membrane in a concentration-dependent manner and more strongly with edited 5-HTR2c-Fl. These results suggest that 5-HTR2c pre-mRNA editing and splicing are entwined processes determining increased 5-HTR2c levels in pathological conditions through a deficit in 5-HTR2c-Tr.

Building a virtual archive using brain architecture and Web 3D to deliver neuropsychopharmacology content over the Internet.

The vast amount of heterogeneous data generated in various fields of neurosciences such as neuropsychopharmacology can hardly be classified using traditional databases. We present here the concept of a virtual archive, spatially referenced over a simplified 3D brain map and accessible over the Internet. A simple prototype (available at http://aquatics.crs4.it/neuropsydat3d) has been realized using current Web-based virtual reality standards and technologies. It illustrates how primary literature or summary information can easily be retrieved through hyperlinks mapped onto a 3D schema while navigating through neuroanatomy. Furthermore, 3D navigation and visualization techniques are used to enhance the representation of brain's neurotransmitters, pathways and the involvement of specific brain areas in any particular physiological or behavioral functions. The system proposed shows how the use of a schematic spatial organization of data, widely exploited in other fields (e.g. Geographical Information Systems) can be extremely useful to develop efficient tools for research and teaching in neurosciences.

Comparison of dietary fiber contents of selected baby foods from two major brands in Canada using three methods.

Of the two major brands of baby foods in Canada, one reports lower dietary fiber values than the other, although the products appear to be similar. To investigate the reasons for this discrepancy, seven selected samples of baby foods from both brands were analyzed for total dietary fiber (TDF) according to the Mongeau (rapid Health Protection Branch; HPB) method. Two cereals were also analyzed by using the Prosky and the Englyst (nonstarch polysaccharide; NSP) methods as an internal check on the methodology as well as a means of investigating the reasons for the discrepancies. The sampling included at least four different lots of each product (cereals, fruits, vegetables, and legumes). Each lot was analyzed individually. The TDF values determined using the rapid HPB method were in agreement with those obtained by other dietary fiber methods. Comparison between manufacturer-reported and measured values showed that the low values reported in brand A products were due, in part, to under-reporting of TDF content: measured TDF values were significantly higher than manufacturer-reported values. For brand B products, the manufacturer-reported and measured TDF values were in general agreement. This shows that a large part of the discrepancy between the two brands was due to methodological problems associated with measuring TDF in brand A. Differences in TDF content were also apparent as shown by the fact that brand A TDF values were consistently lower than those of brand B when both were measured by the same method. The differences in TDF content were not explained by differences in the polysaccharide composition of the fiber residues or by differences in water content. Although the limited number of samples does not allow any general conclusion about the TDF content of specific brands, the results show that formulation and/or manufacturing differences may influence TDF values in processed baby foods.

Differential effects of neurokinin-1 receptor activation in subregions of the periaqueductal gray matter on conditional and unconditional fear behaviors in rats.

Central neurokinin-1 (NK-1) receptors are thought to modulate aversion, whereas the periaqueductal gray matter (PAG) is a common pathway for the integration of fear behaviors. The authors determined whether injection of an NK-1 agonist (GR73632) into subregions of the PAG would alter fear-related behaviors. Behavioral inactivity was increased by GR73632 injected into the caudodorsal PAG or the dorsal raphe. Flight behavior induced by stimulation of the dorsal PAG or by a footshock was decreased after injection of GR73632 into the dorsal PAG. Rats that had 6 pairings of a tone with a footshock after injection of GR73632 into the dorsal PAG displayed more freezing behavior than controls at the beginning of the session. However, there was no change in the shock- or the tone-induced freezing because some GR73632-treated rats, but no controls, froze during the baseline period. It is concluded that NK-1 receptors in the dorsal PAG modulate the unconditional but not the mnemonic aspects of fear behaviors.

Effect of acute, short- and long-term milnacipran administration on rat locus coeruleus noradrenergic and dorsal raphe serotonergic neurons.

The effect of milnacipran on the firing activity of dorsal raphe serotonin (5-HT) neurons and locus coeruleus norepineprine (NE) neurons was assessed using extracellular unitary recording in chloral hydrate anesthetized rats. A 2-day treatment with milnacipran (20 or 60 mg/kg/day, s.c.) markedly decreased the firing rate of NE neurons, and it remained reduced after a 7- or a 14-day treatment. Although the suppressant effect of the alpha2-adrenergic agonist clonidine on the firing rate of NE neurons was markedly reduced following long-term milnacipran (60 mg/kg/day x 14 days, s.c.), that of NE remained unchanged. The firing rate of 5-HT neurons was reduced following a 2-day treatment with milnacipran (20 mg/kg/day, s.c.), but there was a partial recovery after a 7-day treatment (20 mg/kg/day, s.c.) and a complete one after a 14-day treatment (20, 40 or 60 mg/kg/day, s.c.). The suppressant effect of 5-HT and of the 5-HT1A agonist 8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin) on the firing rate of 5-HT neurons was also unaltered after milnacipran (60 mg/kg/day x 14 days, s.c.). The latter milnacipran treatment did not affect the uptake of [3H]5-HT but it inhibited that of [3H]NE by 30% in hippocampal slices. The NE system was thus investigated in an attempt to explain the effects of milnacipran on the firing activity of 5-HT neurons. Acute injection of milnacipran suppressed the firing activity of 5-HT neurons (with an ED50 of 5.7+/-1.5 mg/kg, i.v.), but not in NE-denervated rats. Furthermore, the inhibitory effect of clonidine on 5-HT neuron firing activity was markedly reduced by the long-term milnacipran treatment, whereas the inhibition of electrically evoked release of [3H]NE as well as that of [3H]5-HT produced by the alpha2-adrenoceptor agonist UK 14.304 from preloaded mesencephalic slices containing the dorsal raphe was unaltered. The latter results indicate that the alpha2-adrenergic autoreceptor and heteroreceptor were unaffected in the raphe area by milnacipran. In conclusion, milnacipran had profound effects on the function of 5-HT and NE neurons, and the mechanism by which 5-HT neurons regained their normal firing during milnacipran treatment appeared to implicate the NE system.

Effect of central and peripheral administrations of cholecystokinin-tetrapeptide on panic-like reactions induced by stimulation of the dorsal periaqueductal grey area in the rat.

Administration of cholecystokinin-tetrapeptide (CCK-4) triggers panic attacks in humans, but it is not known whether CCK-4 acts in the brain to produce this effect. Panic-like reactions (flight and tachycardia) induced in rats by injecting D, L-homocysteic acid (DLH) into the dorsal periaqueductal grey area (DPAG), were used as an animal model to investigate this issue. CCK-4 (2 micrograms) infused into the DPAG did not change these panic-like reactions. The DLH-induced tachycardia was prolonged by intracerebroventricular injection of CCK-4 (40 or 4 micrograms); however, the DLH-induced flight behavior was not changed by similar central injections of CCK-4 (40, 4, or 0.4 micrograms). Peripheral injection of t-butoxycarbonyl (BOC)-CCK-4 (40 micrograms) potentiated the flight behavior, but did not alter the tachycardia response. It was concluded that CCK tetrapeptide potentiates panic-like behaviors by acting on a peripheral target or on a circumventricular area of the brain. In contrast, increased brain CCK-4 prolongs tachycardia by acting in the brain at a level distinct from the DPAG.

Effect of imipramine treatments on the 5-HT1A-receptor-mediated inhibition of panic-like behaviours in rats.

The escape behaviour induced in rats by injecting D,L-homocysteic acid (DHL) into the dorsal periaqueductal grey area (DPAG) was used as an animal model of panic attacks to investigate the effect of imipramine, a drug used for the treatment of panic disorder, on the sensitivity of 5-HT1A receptors in the DPAG. Rats given imipramine (10 mg/kg per day SC for 3 weeks or IP for 2-3 days) received 250 nl saline or the 5-HT1A agonist 8-OH-DPAT (8.6 nmol) into the DPAG 10 min before inducing the escape response with DLH. As expected, 8-OH-DPAT produced a marked decrease in the average speed of the DLH-induced flight response. The short-term treatment with imipramine changed neither the DLH-induced escape behaviour nor the effect of prior 8-OH-DPAT administration on this response. In contrast, long-term treatment with imipramine enhanced the 5-HT1A-mediated inhibition, as the decrement in the amplitude of the flight response produced by 8-OH-DPAT was 96% after this treatment compared to 41% in controls. The injection of 8-OH-DPAT also significantly decreased the amplitude of the freezing behaviour observed at the end of the flight response in rats given imipramine for 3 weeks, but not in controls. The long-term imipramine treatment, however, did not significantly decrease the amplitude of DLH-induced flight and freezing behaviours in absence of prior 8-OH-DPAT administration. Finally, 8-OH-DPAT failed to inhibit the DLH-induced flight and freezing behaviours in rats withdrawn from imipramine after long-term treatment (10 mg/kg per day x 21 days). It is suggested that an alteration at the level of the DPAG-5-HT1A receptor system is implicated in the therapeutic and withdrawal effect of imipramine in panic disorder.

The serotonergic and noradrenergic systems of the hippocampus: their interactions and the effects of antidepressant treatments.

Previous reviews have well illustrated how antidepressant treatments can differentially alter several neurotransmitter systems in various brain areas. This review focuses on the effects of distinct classes of antidepressant treatments on the serotonergic and the noradrenergic systems of the hippocampus, which is one of the brain limbic areas thought to be relevant in depression: it illustrates the complexity of action of these treatments in a single brain area. First, the basic elements (receptors, second messengers, ion channels, ...) of the serotonergic and noradrenergic systems of the hippocampus are revisited and compared. Second, the extensive interactions occurring between the serotonergic and the noradrenergic systems of the brain are described. Finally, issues concerning the short- and long-term effects of antidepressant treatments on these systems are broadly discussed. Although there are some contradictions, the bulk of data suggests that antidepressant treatments work in the hippocampus by increasing and decreasing, respectively, serotonergic and noradrenergic neurotransmission. This hypothesis is discussed in the context of the purported function of the hippocampus in the formation of memory traces and emotion-related behaviors.

Importance of cooking temperature and pancreatic amylase in determination of dietary fiber in dried legumes.

Total dietary fiber (TDF) was measured in large lima, roman, black turtle, light red kidney, white navy, pinto, black-eyed, and soya beans and in chick peas by the Mongeau rapid method (A), the Prosky method (B), and the Lee method (C). When the samples were soaked and cooked according to package instructions (gentle boiling, 95 degrees C), TDF values by method A were all within 19.7-22.1%, except for black-eyed beans (9.9%) and chick peas (11.3%) (g/100 g, cooked dry matter). For large lima beans (20.0-21.3%) and soya beans (19.2-19.7%), TDF values by methods A, B, and C were in agreement. For 7 samples, however, TDF values were up to 81% higher by method B (17.4-34.7%) and up to 122% higher by method C (21.1-39.8%) than those by method A (P < or = 0.01). For 6 legumes, TDF values by method C were 15-28% higher (P < or = 0.013) than by method B. White navy beans were analyzed also after different cooking conditions, varying from no cooking to autoclaving for 15 min at 120 degrees C. TDF values by method A were independent from cooking conditions and remained between 20.2 and 22.4%. For navy beans cooked at 95 degrees C, TDF values by method B (up to 34.7 +/- 1.4%) and C (up to 39.8 +/- 0.3%) were unpredictable, but autoclaving at 120 degrees C reduced them to about 22%. Incorporation of a pancreatic amylase in methods B and C consistently decreased the aforementioned analytical discrepancies, as did the absence of cooking. Only autoclaving (for at least 15 min at 120 degrees C) fully restored agreement among methods A-C.

Effects of amylose and wheat bran on the levels of blood serum urea nitrogen (BUN), other blood parameters, growth and fecal characteristics in rats.

A basal diet (containing 20% soybean protein isolate) was supplemented with limiting amino acids (AA, methionine, threonine and tryptophan); wheat bran (WB, 24.3%); amylose (AM, 10%) or a combination of AA and WB or AM to investigate their effects on the levels of blood serum urea nitrogen (BUN), other blood parameters, growth and fecal characteristics in young rats. After 3 weeks feeding, supplementation of the basal diet with AA or WB resulted in improved growth (gain/food ratio) while the addition of AM had no effect on rat growth. Addition of WB to the basal diet had no effect on BUN while the addition of AA and AM caused about 20 and 12% reduction in BUN values, respectively. Rats fed the AA-AM diet had the lowest BUN values (42% lower than those fed the basal diet). Fresh volume, fresh weight, moisture and dry weight of feces in rats fed the WB diets were about four to five fold higher than in those fed the AM diets. The results suggested that BUN may not be a good predictor of growth and protein quality in diets containing AM or a source of dietary fiber (WB).

Effect of long-term administration of antidepressant drugs on the 5-HT3 receptors that enhance the electrically evoked release of [3H]noradrenaline in the rat hippocampus.

The present study investigated the effects of various classes of antidepressant drugs (10 mg/kg per day, s.c. during 21 days) on the electrically evoked release of [3H]noradrenaline and on its modulation by the 5-HT3 receptor agonist 2-methyl-5-hydroxy-tryptamine (2-methyl-5-HT) using preloaded rat hippocampal slices. Treatments with either fluoxetine, a selective serotonin (5-HT) reuptake inhibitor, or moclobemide, a reversible type A monoamine oxidase inhibitor, increased the evoked release of [3H]noradrenaline. These two antidepressant treatments did not change, however, the magnitude of the enhancing effect of 2-methyl-5-HT on the electrically evoked release of [3H]noradrenaline. Desipramine produced a much larger increase of the electrically evoked release of [3H]noradrenaline than fluoxetine or moclobemide, and desensitized the 5-HT3 receptors that modulate this release. Trimipramine, which like desipramine has a tricyclic structure but does not block the reuptake of noradrenaline or that of 5-HT, did not increase the evoked release of [3H]noradrenaline and did not desensitize the 5-HT3 receptors that enhance the release of [3H]noradrenaline. Maprotiline, a selective noradrenaline reuptake inhibitor, did not produce the same changes as desipramine, but maprotiline inhibited noradrenaline reuptake to a lesser extent (50%) than desipramine (80%). These results suggest that the high potency noradrenaline reuptake blocker desipramine desensitizes 5-HT3 receptors modulating [3H]noradrenaline release, but that this effect is not common to all antidepressant drugs.

Comparison and assessment of the difference in total dietary fiber in cooked dried legumes as determined by five methods.

Dried beans (kidney and great northern) and dried peas (chick, green, and yellow) were cooked according to package instructions. Total dietary fiber (TDF) was measured by the Mongeau (AOAC 992.16), Prosky (AOAC 985.29), and Lee (AOAC 991.43) methods (A, B, and C, respectively). Nonstarch polysaccharides (NSP) were measured by the Englyst gas-chromatographic method that included dimethyl sulfoxide treatment, and the lignin measured separately was added to NSP (method D). TDF was also measured by the Li method (E). TDF values ranged from 7 to 48 g/100 g dry weight. Methods B and C gave similar TDF values, which were higher than those from other methods. For kidney beans I, TDF values by methods B and C were up to 2.5 times higher because of the inclusion of starch. The fiber residues from methods A and C contained the same amounts of arabinose, xylose, mannose, galactose, and uronic acid, but glucose was 4 times higher in residue C than in residue A. When alpha-amylase from porcine pancreas was incorporated in methods B or C for 5 samples, the discrepancies among methods A, B, and C were reduced by 60-98%.

Effects of long-term alpha-2 adrenergic antagonists and electroconvulsive treatments on the alpha-2 adrenoceptors modulating serotonin neurotransmission.

Previous results from our laboratory indicate that small doses of the alpha-2 adrenergic agonist clonidine increase serotonin (5-HT) neurotransmission by attenuating the release of endogenous norepinephrine (NE), as a result of the activation of alpha-2 adrenergic autoreceptors on NE neurons, and that high doses decrease 5-HT neurotransmission by activating directly alpha-2 adrenergic heteroreceptors on 5-HT terminals. In addition, we have shown that long-term treatments with a monoamine oxidase inhibitor or a selective NE, but not a 5-HT, reuptake inhibitor abolish the effect of a high dose of clonidine, but not that of a small dose of clonidine. The aim of the present study was to determine whether the alpha-2 adrenergic antagonists idazoxan (10 mg/kg/day x 21 days s.c.) and mianserin (5 mg/kg/day x 21 days s.c.), or electroconvulsive shocks (6 or 7 over a 2-week period) would also affect the alpha-2 adrenoceptors modulating 5-HT neurotransmission in the rat hippocampus. The responsiveness of these hetereceptors was tested in parallel with those of the terminal 5-HT1B autoreceptors and of the postsynaptic 5-HT1A and alpha-2 adrenergic receptors. None of the above treatments altered the responsiveness of the 5-HT1B autoreceptors, as assessed by comparing the differential effectiveness of 1 and 5 Hz electrical stimulations of the 5-HT pathway. Idazoxan and mianserin did not affect the responsiveness of the postsynaptic 5-HT1A and alpha-2 adrenergic receptors as indicated by the unchanged suppressant effects of microiontophoretically applied 5-HT and NE.(ABSTRACT TRUNCATED AT 250 WORDS)