The effect of valproate on the amino acids, monoamines, and kynurenic acid concentrations in brain structures involved in epileptogenesis in the pentylenetetrazol-kindled rats.

BACKGROUND: The study aimed to assess the influence of a single valproate (VPA) administration on inhibitory and excitatory neurotransmitter concentrations in the brain structures involved in epileptogenesis in pentylenetetrazol (PTZ)-kindled rats. METHODS: Adult, male Wistar rats were kindled by repeated intraperitoneal (ip) injections of PTZ at a subconvulsive dose (30 mg/kg, three times a week). Due to the different times required to kindle the rats (18-22 injections of PTZ), a booster dose of PTZ was administrated 7 days after the last rats were kindled. Then rats were divided into two groups: acute administration of VPA (400 mg/kg) or saline given ip. The concentration of amino acids, kynurenic acid (KYNA), monoamines, and their metabolites in the prefrontal cortex, hippocampus, amygdala, and striatum was assessed by high-pressure liquid chromatography (HPLC). RESULTS: It was found that a single administration of VPA increased the gamma-aminobutyric acid (GABA), tryptophan (TRP), 5-hydroxyindoleacetic acid (5-HIAA), and KYNA concentrations and decreased aspartate (ASP) levels in PTZ-kindled rats in the prefrontal cortex, hippocampus, amygdala and striatum. CONCLUSIONS: Our results indicate that a single administration of VPA in the PTZ-kindled rats restored proper balance between excitatory (decreasing the level of ASP) and inhibitory neurotransmission (increased concentration GABA, KYNA) and affecting serotoninergic neurotransmission in the prefrontal cortex, hippocampus, amygdala, and striatum.

The role of the dorsal hippocampus in resistance to the development of posttraumatic stress disorder-like behaviours.

This study aimed to determine the activity of the dorsal hippocampus (dHIP) in resistance to the development of posttraumatic stress disorder (PTSD)-like behaviours. Rats were divided into resistant, PTSD(-), and susceptible, PTSD(+) groups based on the time spent in the central area in an open field test and freezing duration during exposure to an aversive context one week after stress experience (electric foot shock). The PTSD(-) rats, compared to the PTSD(+) group, had an increased concentration of corticosterone in plasma and changes in the activity of the dHIP, specifically, increased c-Fos expression in the dentate gyrus (DG) and increased Neuroligin-2 (marker of GABAergic neurotransmission) expression in the DG and CA3 area of the dHIP. Moreover, in the hippocampus, the PTSD(-) group showed decreased mRNA expression for corticotropin-releasing factor receptors type 1 and 2, increased mRNA expression for orexin receptor type 1, and decreased miR-9 and miR-34c levels compared with the PTSD(+) group. This study may suggest that the increase in GABA signalling in the hippocampus attenuates the activity of the CRF system and enhances the function of the orexin system. Moreover, decreased expression of miR-34c and miR-9 could facilitate fear extinction and diminishes the anxiety response. These effects may lead to an anxiolytic-like effect and improve resistance to developing PTSD-like behaviours.

The effects of the recurrent social isolation stress on fear extinction and dopamine D2 receptors in the amygdala and the hippocampus.

BACKGROUND: The present study assessed the influence of recurrent social isolation stress on the aversive memory extinction and dopamine D2 receptors (D2R) expression in the amygdala and the hippocampus subnuclei. We also analyzed the expression of epigenetic factors potentially associated with fear extinction: miRNA-128 and miRNA-142 in the amygdala. METHODS: Male adult fear-conditioned rats had three episodes of 48 h social isolation stress before each fear extinction session in weeks intervals. Ninety minutes after the last extinction session, the D2R expression in the nuclei of the amygdala and the hippocampus (immunocytochemical technique), and mRNA levels for D2R in the amygdala were assessed (PCR). Moreover, we evaluated the levels of miRNA-128 and miRNA-142 in the amygdala. RESULTS: It was found that recurrent social isolation stress decreased the fear extinction rate. The extinguished isolated rats were characterized by higher expression of D2R in the CA1 area of the hippocampus compared to the extinguished and the control rats. In turn, the isolated group presented higher D2R immunoreactivity in the CA1 area compared to the extinguished, the control, and the extinguished isolated animals. Moreover, the extinguished animals had higher expression of D2R in the central amygdala than the control and the extinguished isolated rats. These changes were accompanied by the increase in miRNA-128 level in the amygdala in the extinguished isolated rats compared to the control, the extinguished, and the isolated rats. Moreover, the extinguished rats had lower expression of miRNA-128 compared to the control and the isolated animals. CONCLUSIONS: Our results suggest that social isolation stress impairs aversive memory extinction and coexists with changes in the D2R expression in the amygdala and hippocampus and increased expression of miRNA-128 in the amygdala.

Molecular pattern of a decrease in the rewarding effect of cocaine after an escalating-dose drug regimen.

BACKGROUND: Long-term cocaine exposure leads to dysregulation of the reward system and initiates processes that ultimately weaken its rewarding effects. Here, we studied the influence of an escalating-dose cocaine regimen on drug-associated appetitive behavior after a withdrawal period, along with corresponding molecular changes in plasma and the prefrontal cortex (PFC). METHODS: We applied a 5 day escalating-dose cocaine regimen in rats. We assessed anxiety-like behavior at the beginning of the withdrawal period in the elevated plus maze (EPM) test. The reinforcement properties of cocaine were evaluated in the Conditioned Place Preference (CPP) test along with ultrasonic vocalization (USV) in the appetitive range in a drug-associated context. We assessed corticosterone, proopiomelanocortin (POMC), ß-endorphin, CART 55-102 levels in plasma (by ELISA), along with mRNA levels for D2 dopaminergic receptor (D2R), κ-receptor (KOR), orexin 1 receptor (OX1R), CART 55-102, and potential markers of cocaine abuse: miRNA-124 and miRNA-137 levels in the PFC (by PCR). RESULTS: Rats subjected to the escalating-dose cocaine binge regimen spent less time in the cocaine-paired compartment, and presented a lower number of appetitive USV episodes. These changes were accompanied by a decrease in corticosterone and CART levels, an increase in POMC and ß-endorphin levels in plasma, and an increase in the mRNA for D2R and miRNA-124 levels, but a decrease in the mRNA levels for KOR, OX1R, and CART 55-102 in the PFC. CONCLUSIONS: The presented data reflect a part of a bigger picture of a multilevel interplay between neurotransmitter systems and neuromodulators underlying processes associated with cocaine abuse.

Neurobiological Basis of Increased Risk for Suicidal Behaviour.

According to the World Health Organization (WHO), more than 700,000 people die per year due to suicide. Suicide risk factors include a previous suicide attempt and psychiatric disorders. The highest mortality rate in suicide worldwide is due to depression. Current evidence suggests that suicide etiopathogenesis is associated with neuroinflammation that activates the kynurenine pathway and causes subsequent serotonin depletion and stimulation of glutamate neurotransmission. These changes are accompanied by decreased BDNF (brain-derived neurotrophic factor) levels in the brain, which is often linked to impaired neuroplasticity and cognitive deficits. Most suicidal patients have a hyperactive hypothalamus-pituitary-adrenal (HPA) axis. Epigenetic mechanisms control the above-mentioned neurobiological changes associated with suicidal behaviour. Suicide risk could be attenuated by appropriate psychological treatment, electroconvulsive treatment, and drugs: lithium, ketamine, esketamine, clozapine. In this review, we present the etiopathogenesis of suicide behaviour and explore the mechanisms of action of anti-suicidal treatments, pinpointing similarities among them.

Sex-Related Predisposition to Post-Traumatic Stress Disorder Development-The Role of Neuropeptides.

Post-traumatic stress disorder (PTSD) is characterized by re-experiencing a traumatic event, avoidance, negative alterations in cognitions and mood, hyperarousal, and severe functional impairment. Women have a two times higher risk of developing PTSD than men. The neurobiological basis for the sex-specific predisposition to PTSD might be related to differences in the functions of stress-responsive systems due to the interaction between gonadal hormones and stress peptides such as corticotropin-releasing factor (CRF), orexin, oxytocin, and neuropeptide Y. Additionally, in phases where estrogens levels are low, the risk of developing or exacerbating PTSD is higher. Most studies have revealed several essential sex differences in CRF function. They include genetic factors, e.g., the CRF promoter contains estrogen response elements. Importantly, sex-related differences are responsible for different predispositions to PTSD and diverse treatment responses. Fear extinction (the process responsible for the effectiveness of behavioral therapy for PTSD) in women during periods of high endogenous estradiol levels (the primary form of estrogens) is reportedly more effective than in periods of low endogenous estradiol. In this review, we present the roles of selected neuropeptides in the sex-related predisposition to PTSD development.

Individual susceptibility or resistance to posttraumatic stress disorder-like behaviours.

The aim of this study was to explore the neurobiological background of individual susceptibility and resistance to the development of posttraumatic stress disorder (PTSD)-like behaviours. Rats were divided into susceptible, PTSD(+), and resistant, PTSD(-), groups based on freezing duration during exposure to aversive context and the time spent in the central area in open field test one week after threefold stress experience (modified single prolonged stress). PTSD(-) rats showed increased concentrations of corticosterone in plasma and changes in GAD67 expression: decreased in the infralimbic cortex (IL) and increased in the lateral amygdala (LA), dentate gyrus (DG), and CA1 area of the hippocampus. Moreover, in this group, we found an increase in the number of CRF-positive nuclei in the parvocellular neurons of the paraventricular hypothalamic nucleus (pPVN). The PTSD(+) group, compared to PTSD(-) rats, had decreased concentrations of corticosterone in plasma and reduced CRF expression in the pPVN, higher CRF expression in the CA1, increased expression of CRF-positive nuclei and GR receptors in the CA3 area of the hippocampus, and increased expression of GR receptors in the DG and the central amygdala (CeA). Biochemical analysis showed higher concentrations of noradrenaline, glutamic acid in the dorsal hippocampus and amygdala and lower levels of dopamine and its metabolites in the amygdala of the PTSD(+) group than in the PTSD(-) group. The study revealed different behavioural and biochemical profiles of PTSD(+) and PTSD(-) rats and suggested that individual differences in hypothalamic-pituitary-adrenal (HPA) axis activity may determine hippocampal- and amygdala-dependent memory and fear processing.

Stress changes amphetamine response, D2 receptor expression and epigenetic regulation in low-anxiety rats.

The aim of this study was to assess the influence of chronic restraint stress on amphetamine (AMPH)-related appetitive 50-kHz ultrasonic vocalisations (USVs) in rats differing in freezing duration in a contextual fear test (CFT), i.e. HR (high-anxiety responsive) and LR (low-anxiety responsive) rats. The LR and the HR rats, previously exposed to an AMPH binge experience, differed in sensitivity to AMPH's rewarding effects, measured as appetitive vocalisations. Moreover, chronic restraint stress attenuated AMPH-related appetitive vocalisations in the LR rats but had no influence on the HR rats' behaviour. To specify, the restraint LR rats vocalised appetitively less in the AMPH-associated context and after an AMPH challenge than the control LR rats. This phenomenon was associated with a decrease in the mRNA level for D2 dopamine receptor in the amygdala and its protein expression in the basal amygdala (BA) and opposite changes in the nucleus accumbens (NAc) - an increase in the mRNA level for D2 dopamine receptor and its protein expression in the NAc shell, compared to control conditions. Moreover, we observed that chronic restraint stress influenced epigenetic regulation in the LR and the HR rats differently. The contrasting changes were observed in the dentate gyrus (DG) of the hippocampus - the LR rats presented a decrease, but the HR rats showed an increase in H3K9 trimethylation. The restraint LR rats also showed higher miR-494 and miR-34c levels in the NAc than the control LR group. Our study provides behavioural and biochemical data concerning the role of differences in fear-conditioned response in stress vulnerability and AMPH-associated appetitive behaviour. The LR rats were less sensitive to the rewarding effects of AMPH when previously exposed to chronic stress that was accompanied by changes in D2 dopamine receptor expression and epigenetic regulation in mesolimbic areas.

The role of UCH-L1, MMP-9, and GFAP as peripheral markers of different susceptibility to seizure development in a preclinical model of epilepsy.

In our study, we assessed the potency of the brain-derived proteins ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), matrix metalloproteinase 9 (MMP-9), glial fibrillary acidic protein (GFAP) and the immune activation indicators interleukin 1ß (IL-1ß) and interleukin 6 (IL-6) as peripheral biomarkers of different susceptibilities to kindling in a preclinical model. We observed increased plasma UCH-L1 levels in kindled vs. control animals. Furthermore, MMP-9 and IL-1ß concentrations were the lowest in rats resistant to kindling. In summary, UCH-L1 is an indicator of neuronal loss and BBB disruption after seizure. MMP-9 and IL-1ß may indicate resistance to kindling. UCH-L1, MMP-9 and IL-1ß may have utility as peripheral biomarkers with translational potency in the clinic.

The effect of a corticotropin-releasing factor receptor 1 antagonist on the fear conditioning response in low- and high-anxiety rats after chronic corticosterone administration.

This study aimed to test the hypothesis that high-anxiety (HR) rats are more sensitive to the effects of chronic corticosterone administration and antalarmin (corticotropin-releasing factor (CRF) receptor 1, CRF1 antagonist) injections than low-anxiety (LR) rats, and this effect is accompanied by changes in CRF system activity in brain regions involved in the control of emotions and the hypothalamic-pituitary-adrenal (HPA) axis. Male rats were divided into LR (n = 25) and HR (n = 30) groups according to the duration of conditioned freezing in a contextual fear test. Chronic corticosterone administration (by injection, 20 mg/kg) for 21 d (except weekends) increased freezing duration and number of GR (glucocorticoid receptor)-immunoreactive nuclei in the basal amygdala (BA) and decreased GR-immunoreactive nuclei in the infralimbic cortex (IL), dentate gyrus (DG), and CA3 area, only in the HR group. Moreover, in this group, corticosterone administration decreased number of CRF-immunoreactive neurons of the parvocellular paraventricular hypothalamic nucleus (pPVN), DG, and CA1. Antalarmin (10 mg/kg, i.p., 2 injections) significantly attenuated conditioned fear responses, increased plasma corticosterone concentration, and decreased GR-immunoreactive nuclei in the BA, only in the HR group. Moreover, in this group, antalarmin increased number of GR-immunoreactive nuclei in the IL, DG, and CA3 and increased number of CRF-immunoreactive cells in the pPVN, DG, and CA1. Hence, antalarmin attenuated the fear response and restored HPA axis function in HR rats, which were more sensitive to corticosterone exposure. These data suggest that individual differences in central local CRF system activity may determine the neurobiological mechanisms related to mood and emotional disorders.

A rat model to study maternal depression during pregnancy and postpartum periods, its comorbidity with cardiovascular diseases and neurodevelopmental impact in the offspring.

This study aimed to develop an animal model of human depression during pregnancy and lactation to examine the effect of maternal, perinatal depression on offspring development. Maternal depression during pregnancy affects up to 20% of women and is a risk factor for both the developmental and long-term health issues. It is often comorbid with the cardiovascular disease (CVD) that affects the uteroplacental circulation and impacts offspring development. More than half of the expecting mothers with depression use antidepressants that cross the placenta and may interfere with the neurodevelopmental programming. Thus, depressed pregnant mothers face a difficult choice whether "to use or not to use" antidepressant therapy, since both untreated depression and antenatal antidepressant exposure present increased risks of neurodevelopmental pathologies. The ongoing clinical debate presents inconclusive data, while the existing animal models of maternal depression do not include early gestational periods, and, do not monitor depressive-like behavior nor address the cardiovascular abnormalities. The presented model includes pregestational depressive behavior extending into pregnancy and lactation, periods that have not been previously examined. Rat dams exposed to pre-gestational chronic mild stress (CMS) developed a sustained decrease in self-grooming behavior, correlated with hormonal, behavioral, and cardiac changes persisting through the postpartum period. Preliminary data indicate neurodevelopmental delays, behavioral and cardiac abnormalities, and altered levels of both the brain and the heart markers in the offspring of stressed dams. Furthermore, the preliminary data predict that maternal pregnancy during the perinatal period is likely to impact the neurodevelopmental process in a sex-dependent manner. Thus the presented here model (PG-LAC CMS) fulfills both the face and the construct validity criteria for maternal stress-induced depression during pregnancy and postpartum that may facilitate further studies of the relative risks of untreated vs. antidepressant-treated maternal depression during pregnancy to the mother and her offspring.

Differences in the dopaminergic reward system in rats that passively and actively behave in the Porsolt test.

The aim of the study was to assess appetitive responses and central dopaminergic neurotransmission in passive and active rats divided according to their immobility time in the Porsolt swim test and exposed to restraint stress. Passive rats had more episodes of appetitive 50-kHz ultrasonic vocalization (USV) during rat encounter after social isolation and spent significantly more time in the amphetamine-associated context in conditioned place preference test, compared to active rats. Restraint stress decreased sucrose preference, but increased appetitive vocalization and reinforced the conditioned place preference only in passive animals that was associated with increased dopamine concentration in the amygdala. Restraint stress increased also the level of Cocaine- and Amphetamine Regulated Transcript (CART) peptide, a neuromodulator linked to dopamine neurotransmission, in the central nucleus of amygdala, while decreasing it the nucleus accumbens shell in passive rats. In the parvocellular region of paraventricular nucleus of the hypothalamus passive animals had a higher expression of CART compared to passive restraint rats and active control rats. The obtained results show that active and passive rats in the Porsolt test differ significantly in response to appetitive stimuli, which can be additionally changed under stress conditions. The underlying mechanisms are probably associated with differences in dopaminergic activity and CART signaling in reward system.

Dopaminergic system activity under stress condition - seeking individual differences, preclinical studies. / Funkcjonowanie ukladu dopaminergicznego w warunkach stresu ­ poszukiwanie podstaw róznic indywidualnych, badania przedkliniczne.

Dopaminergic system activity in limbic structures (reward system) is related to motivational processes and adaptation to changing environmental conditions. Stress conditions can cause dopaminergic dysfunction, reduce motivational processes and induce compensatory drug use. The susceptibility to stress is characterized by individual variability. Psychostimulants such as cocaine, amphetamine and its derivatives act as positive reinforcers, affecting mood changes. Prolonged use of psychoactive substances can cause persistent plastic changes in the limbic system (disruption of neurogenesis, neurons atrophy), resulting in addictions or other forms of psychopathology like mood disorders. One of the reason is dysregulation of the dopaminergic system and dysfunction of local dopamine release in the nucleus accumbens. Stress factors also inhibit neuronal plasticity. In turn, antidepressants may increase brain-derived neurotrophic factor (BDNF) and TrkB receptors expression and improve neuronal proliferation, restoring proper functioning of the limbic regions. An important manifestation of the distinct functioning of the dopaminergic mesolimbic system is the difference between the sexes and the aging process. Epidemiological studies indicate that depression, anxiety disorders, and other emotional disorders often accompany drug abuse. The search for neurobiological basis of affective disorders and identification of factors, including epigenetic ones (interdependence of genetic and environmental factors), associated with different susceptibility to stress and predisposition to addiction to psychoactive substances is currently being carried out by many researches. Understanding the neurobiological factors of individual differences related to susceptibility to psychostimulants may aid in developing future therapies adapted to the patient's needs and more effective treatment of addiction.

The amphetamine-associated context exerts a stronger motivational effect in low-anxiety rats than in high-anxiety rats.

This study used the conditioned place preference test to explore the effects of subchronic amphetamine administration on drug-associated cues in rats with different emotional reactivity. We also examined the changes in markers of dopaminergic activity in brain regions in response to the amphetamine-paired context, after a withdrawal period preceded by subchronic amphetamine treatment. We used low-anxiety (LR) and high-anxiety (HR) rats, which are known to exhibit distinct levels of susceptibility to amphetamine. Compared to HR rats, LR rats spent significantly more time in the amphetamine-paired compartment after the withdrawal period preceded by subchronic amphetamine treatment. Compared to HR control rats, LR control rats showed higher expression of the D1 receptor in the nucleus accumbens core (NAC core) and basolateral amygdala and higher expression of the D2 receptor in the NAC core. After the amphetamine treatment and withdrawal period, the LR rats showed higher D1 receptor expression in the NAC core, an increased level of homovanilic acid (HVA) in the prefrontal cortex, the NAC and the central amygdala than HR rats, as well as lower D2 receptor expression in the NAC core and the amygdala than LR control rats. These results indicate that the differences in the activity of the dopaminergic mesolimbic system in the HR and LR rats are maintained and even enhanced after a multi-day break in the use of the drug, indicating the occurrence of sensitisation. These findings show that the innate reactivity of the limbic dopaminergic innervations, dependent on the level of emotional reactivity, may significantly and chronically modify the development and maintenance of sensitisation to amphetamine.

Low-anxiety rats are more sensitive to amphetamine in comparison to high-anxiety rats.

This study utilised the two injection protocol of sensitisation (TIPS) and the conditioned place preference test to validate and extend previous findings on the effects of amphetamine on positive reinforcement-related 50 kHz ultrasonic vocalisation (USV) in rats. We also examined changes in the expression of c-Fos and the NMDA receptor 2B (GluN2B) subunit, markers of neuronal activity and plasticity, in brain regions of rats in response to TIPS. We used low anxiety-responsive (LR) and high anxiety-responsive (HR) rats, which are known to exhibit different fear-conditioned response strengths, different susceptibilities to amphetamine in the TIPS procedure and different amphetamine-dependent 50 kHz USV responses. The LR rats, compared to the HR rats, not only vocalised much more intensely but also spent significantly more time in the amphetamine-paired compartment. After the second dose of amphetamine, the LR rats exhibited more c-Fos and GluN2B activation in layers II and III of the M1/M2 motor cortex area and prefrontal cortex (PRE, PRL, IL) and also presented with more GluN2B activation in the basal amygdala. These data reveal that HR and LR rats exhibit different levels of reactivity in the cortical-limbic pathway, which controls reward-related motivational processes. These findings contribute to the general hypothesis that heterogeneity in emotional processes is one of the causes of sensitisation to amphetamine and drug addiction.

The co-expression of GluN2B subunits of the NMDA receptors and glucocorticoid receptors after chronic restraint stress in low and high anxiety rats.

The aim of this study was to assess the mechanisms underlying behavioural differences between high- (HR) and low- (LR) anxiety rats, selected according to their behaviour in the contextual fear test (i.e., the duration of the freezing response was used as a discriminating variable), after a chronic restraint procedure (21days, 3h daily). We analysed the expression of the GluN2B subunits of the NMDA and glucocorticoid receptors (GRs) in selected brain structures (immunofluorescence). Following chronic restraint stress in the HR rats, we observed a decrease in the expression of the GRs and GluN2B subunits of the NMDA receptor in the prefrontal cortical areas and the hippocampus compared to the HR-control and the LR-restraint groups. These effects coincided with an increase in passive depressive-like behaviour in the Porsolt test of the HR rats. Moreover, in the hippocampus, the HR-restraint animals demonstrated decreased glutamate levels and a decreased glutamate/glutamine ratio compared to the LR-restraint rats. Furthermore, the HR-restraint group had increased GRs/GluN2B subunits colocalisation in the basolateral amygdala (BLA) compared to the HR-control and the LR-restraint rats. The present results suggest that in HR rats exposed to chronic restraint stress, the hippocampal and cortical glutamatergic system components are changed. These effects could have a negative influence on the feedback mechanisms regulating the hypothalamic-pituitary-adrenal axis as well as on the behavioural processes expressed as depressive-like symptoms.

Behavioral effects and CRF expression in brain structures of high- and low-anxiety rats after chronic restraint stress.

The aim of our study was to investigate the influence of chronic restraint stress (5 weeks, 3h/day) on behavior and central corticotropin-releasing factor (CRF) expression in rats selected for high (HR) and low anxiety (LR). The conditioned freezing response was used as a discriminating variable. Moreover, we assessed the influence of acute restraint on CRF expression in the brain in HR and LR rats. We found that chronic restraint induced symptoms of anhedonia (decreased consumption of 1% sucrose solution) in HR rats. In addition, HR restraint rats showed an increased learned helplessness behavior (immobility time in the Porsolt test) as well as neophobia in the open field test vs. LR restraint and HR control rats. These behavioral changes were accompanied by a decreased expression of CRF in the paraventricular nucleus of the hypothalamus (pPVN) and the dentate gyrus of the hippocampus (DG) compared to the HR control and LR restraint rat groups, respectively. The acute restraint condition increased the expression of CRF in the pPVN of HR rats compared to the HR control group, and enhanced the expression of CRF in the CA1 area and DG of LR restraint animals compared to the HR restraint and LR control rats, respectively. The present results indicate that chronic restraint stress in high anxiety rats attenuated CRF expression in the pPVN and DG, which was probably due to detrimental actions on the hippocampus-hypothalamus-pituitary-adrenal gland feedback mechanism, thus modulating the stress response and inducing anhedonia and depressive-like symptoms.

GABAergic control of the activity of the central nucleus of the amygdala in low- and high-anxiety rats.

The aim of this study was to examine the role of GABAergic neurotransmission in amygdala nuclei in low- (LR) and high-anxiety (HR) rats after repeated corticosterone administration and acute injection of the benzodiazepine midazolam. The animals were divided into LR and HR groups based on the duration of their conditioned freezing in a contextual fear test (CFT). Repeated daily administration of corticosterone (20 mg/kg s.c.) for 21 injections increased anxiety-like behavior in the open field and reduced body weight in both the LR and HR groups. These effects of corticosterone administration were more pronounced in the HR group. Moreover, in the HR group, chronic corticosterone administration increased the duration of freezing in the CFT test compared with the appropriate control group and treated LR rats. The behavioral effects in HR rats were accompanied by an increase in the expression of c-Fos in the lateral (LA) and central (CeA) nuclei of the amygdala and by a decrease in GABA-A alpha-2 subunit density in the CeA. Acute midazolam administration significantly attenuated the neophobia and conditioned fear responses, decreased c-Fos expression in the LA and CeA, and increased alpha-2 subunit density in the CeA only in the HR group. These studies have shown that HR rats are more susceptible to the anxiogenic effects of chronic corticosterone administration, which are associated with the attenuation of GABAergic control over the amygdala output that controls emotional responses. The current data may increase understanding of the neurobiological mechanisms responsible for individual differences in the psychopathological processes induced by repeated administration of high doses of glucocorticoids or by elevated levels of these hormones, which are associated with chronic stress and affective pathology.

Chronic restraint increases apoptosis in the hippocampus of rats with high responsiveness to fear stimuli.

This study examined the effects of chronic restraint stress and corticosterone treatment on the apoptosis-related processes in the dentate gyrus of the rat hippocampus. This study compared high (HR) and low anxiety rats (LR) (as defined by their behaviour during the contextual fear test, i.e., the duration of a freezing response was the discriminating variable). The results demonstrate that chronic restraint stress increased the number of caspase-3 immunoreactive cells in the HR group, whereas repeated corticosterone treatment increased the number of caspase-3 immunoreactive cells in both the HR and LR groups. This finding suggests that higher susceptibility to fear stimuli predisposes rats to increased apoptosis in the hippocampus after exposure to chronic stressors. This new animal model of HR and LR rats can be used to study the mechanisms underlying the relationship between higher levels of anxiety and greater vulnerability to stress.

Midazolam treatment before re-exposure to contextual fear reduces freezing behavior and amygdala activity differentially in high- and low-anxiety rats.

The aim of this study was to examine the effects of benzodiazepine (midazolam) administration on rat conditioned fear responses and on local brain activity (c-Fos and CRF expressions) of low- (LR) and high- (HR)anxiety rats after the first and second contextual fear test sessions. The animals were divided into LR and HR groups based on the duration of their conditioned freezing response in the first contextual fear test. The fear-re-conditioned LR and HR animals (28 days later) had increased freezing durations compared with those durations during the first conditioned fear test. These behavioral effects were accompanied by increased c-Fos expression in the medial amygdala (MeA), the basolateral amygdala (BLA), and the paraventricular hypothalamic nuclei and elevated CRF expression in the MeA. All these behavioral and immunochemical effects of fear re-conditioning were stronger in the LR group compared with the effects in the HR group. Moreover, in the LR rats, the re-conditioning led to decreased CRF expression in the primary motor cortex (M1) and to increased CRF expression in the BLA. The pretreatment of rats with midazolam before the second exposure to the aversive context significantly attenuated the conditioned fear response, lowered the serum corticosterone concentration, decreased c-Fos and CRF expressions in the MeA and in the BLA, and increased CRF complex density in M1 area only in the LR group. These studies have demonstrated that LR rats are more sensitive to re-exposure to fear stimuli and that midazolam pretreatment was associated with modified brain activity in the amygdala and in the prefrontal cortex in this group of animals. The current data may facilitate a better understanding of the neurobiological mechanisms responsible for individual differences in the psychopathological processes accompanying some anxiety disorders characterized by stronger reactivity to re-exposure to stressful challenges, e.g., posttraumatic stress disorder.