High Anxiety Rats Research Articles

Wheel-Running Exercise Alleviates Anxiety-Like Behavior via Down-Regulating S-Nitrosylation of Gephyrin in the Basolateral Amygdala of Male Rats.

Physical exercise has beneficial effect on anxiety disorders, but the underlying molecular mechanism remains largely unknown. Here, it is demonstrated that physical exercise can downregulate the S-nitrosylation of gephyrin (SNO-gephyrin) in the basolateral amygdala (BLA) to exert anxiolytic effects. It is found that the level of SNO-gephyrin is significantly increased in the BLA of high-anxiety rats and a downregulation of SNO-gephyrin at cysteines 212 and 284 produced anxiolytic effect. Mechanistically, inhibition of SNO-gephyrin by either Cys212 or Cys284 mutations increased the surface expression of GABAAR γ2 and the subsequent GABAergic neurotransmission, exerting anxiolytic effect in male rats. On the other side, overexpression of neuronal nitric oxide synthase in the BLA abolished the anxiolytic-like effects of physical exercise. This study reveals a key role of downregulating SNO-gephyrin in the anxiolytic effects of physical exercise, providing a new explanation for protein post-translational modifications in the brain after exercise.

Regulation of anxiety-like behaviors by S-palmitoylation and S-nitrosylation in basolateral amygdala

Protein posttranslational modification regulates synaptic protein stability, sorting and trafficking, and is involved in emotional disorders. Yet the molecular mechanisms regulating emotional disorders remain unelucidated. Here we report unknown roles of protein palmitoylation/nitrosylation crosstalk in regulating anxiety-like behaviors in rats. According to the percentages of open arm duration in the elevated plus maze test, the rats were divided into high-, intermediate- and low-anxiety groups. The palmitoylation and nitrosylation levels were detected by acyl-biotin exchange assay, and we found low palmitoylation and high nitrosylation levels in the basolateral amygdala (BLA) of high-anxiety rats. Furthermore, we observed that 2-bromopalmitate (2-BP), a palmitoylation inhibitor, induced anxiety-like behaviors, accompanied with decreased amplitude and frequency of mEPSCs and mIPSCs in the BLA. Additionally, we also found that inhibiting nNOS activity with 7-nitroindazole (7-NI) in the BLA caused anxiolytic effects and reduced the synaptic transmission. Interestingly, diazepam (DZP) rapidly elevated the protein palmitoylation level and attenuated the protein nitrosylation level in the BLA. Specifically, similar to DZP, the voluntary wheel running exerted DZP-like anxiolytic action, and induced high palmitoylation and low nitrosylation levels in the BLA. Lastly, blocking the protein palmitoylation with 2-BP induced an increase in protein nitrosylation level, and attenuating the nNOS activity by 7-NI elevated the protein palmitoylation level. Collectively, these results show a critical role of protein palmitoylation/nitrosylation crosstalk in orchestrating anxiety behavior in rats, and it may serve as a potential target for anxiolytic intervention.

Unveiling the Link: Exploring Mitochondrial Dysfunction as a Probable Mechanism of Hepatic Damage in Post-Traumatic Stress Syndrome.

PTSD is associated with disturbed hepatic morphology and metabolism. Neuronal mitochondrial dysfunction is considered a subcellular determinant of PTSD, but a link between hepatic mitochondrial dysfunction and hepatic damage in PTSD has not been demonstrated. Thus, the effects of experimental PTSD on the livers of high anxiety (HA) and low anxiety (LA) rats were compared, and mitochondrial determinants underlying the difference in their hepatic damage were investigated. Rats were exposed to predator stress for 10 days. Then, 14 days post-stress, the rats were evaluated with an elevated plus maze and assigned to HA and LA groups according to their anxiety index. Experimental PTSD caused dystrophic changes in hepatocytes of HA rats and hepatocellular damage evident by increased plasma ALT and AST activities. Mitochondrial dysfunction was evident as a predominance of small-size mitochondria in HA rats, which was positively correlated with anxiety index, activities of plasma transaminases, hepatic lipids, and negatively correlated with hepatic glycogen. In contrast, LA rats had a predominance of medium-sized mitochondria. Thus, we show links between mitochondrial dysfunction, hepatic damage, and heightened anxiety in PTSD rats. These results will provide a foundation for future research on the role of hepatic dysfunction in PTSD pathogenesis.

Anxiety-related activity of ventral hippocampal interneurons.

Anxiety is an aversive mood reflecting the anticipation of potential threats. The ventral hippocampus (vH) is a key brain region involved in the genesis of anxiety responses. Recent studies have shown that anxiety is mediated by the activation of vH pyramidal neurons targeting various limbic structures. Throughout the cortex, the activity of pyramidal neurons is controlled by GABA-releasing inhibitory interneurons and the GABAergic system represents an important target of anxiolytic drugs. However, how the activity of vH inhibitory interneurons is related to different anxiety behaviours has not been investigated so far. Here, we integrated in vivo electrophysiology with behavioural phenotyping of distinct anxiety exploration behaviours in rats. We showed that pyramidal neurons and interneurons of the vH are selectively active when animals explore specific compartments of the elevated-plus-maze (EPM), an anxiety task for rodents. Moreover, rats with prior goal-related experience exhibited low-anxiety exploratory behaviour and showed a larger trajectory-related activity of vH interneurons during EPM exploration compared to high anxiety rats. Finally, in low anxiety rats, trajectory-related vH interneurons exhibited opposite activity to pyramidal neurons specifically in the open arms (i.e. more anxiogenic) of the EPM. Our results suggest that vH inhibitory micro-circuits could act as critical elements underlying different anxiety states.

Paradoxical Anxiety Level Reduction in Animal Chronic Stress: A Unique Role of Hippocampus Neurobiology.

A paradoxical reduction in anxiety levels in chronic predator stress paradigm (PS) in Sprague–Dawley rats has recently been shown in previous works. In this paper, we studied the possible neurobiological mechanism of this phenomenon. We segregated PS-exposed Sprague–Dawley rats into the high- and low-anxiety phenotypes. The long-lasting effects of PS on corticosterone levels, blood flow speed in the carotid arteries, diffusion coefficient, and 1H nuclear magnetic resonance spectra in the hippocampus were compared in the high-anxiety and low-anxiety rats. In addition, we evaluated the gene BDNF expression in the hippocampus which is considered to be a main factor of neuroplasticity. We demonstrated that in low-anxiety rats, the corticosterone level was decreased and carotid blood flow speed was increased. Moreover, in the hippocampus of low-anxiety rats compared to the control group and high-anxiety rats, the following changes were observed: (a) a decrease in N-acetyl aspartate levels with a simultaneous increase in phosphoryl ethanol amine levels; (b) an increase in lipid peroxidation levels; (c) a decrease in apparent diffusion coefficient value; (d) an increase in BDNF gene expression. Based on these findings, we proposed that stress-induced anxiety reduction is associated with the elevation of BDNF gene expression directly. Low corticosterone levels and a rise in carotid blood flow speed might facilitate BDNF gene expression. Meanwhile, the decrease in apparent diffusion coefficient value and decrease in N-acetyl aspartate levels, as well as an increase in the lipid peroxidation levels, in the hippocampus possibly reflected destructive changes in the hippocampus. We suggested that in Sprague–Dawley rats, these morphological alterations might be considered as an impetus for further increase in neuroplasticity in the hippocampus.

A Rat Model of Post-Traumatic Stress Syndrome Causes Phenotype-Associated Morphological Changes and Hypofunction of the Adrenal Gland.

Background: Rats exposed to chronic predator scent stress mimic the phenotype of complex post-traumatic stress disorder (PTSD) in humans, including altered adrenal morphology and function. High- and low-anxiety phenotypes have been described in rats exposed to predator scent stress (PSS). This study aimed to determine whether these high- and low-anxiety phenotypes correlate with changes in adrenal histomorphology and corticosteroid production. Methods: Rats were exposed to PSS for ten days. Thirty days later, the rats’ anxiety index (AI) was assessed with an elevated plus-maze test. Based on differences in AI, the rats were segregated into low- (AI ≤ 0.8, n = 9) and high- (AI > 0.8, n = 10) anxiety phenotypes. Plasma corticosterone (CORT) concentrations were measured by ELISA. Adrenal CORT, desoxyCORT, and 11-dehydroCORT were measured by high-performance liquid chromatography. After staining with hematoxylin and eosin, adrenal histomorphometric changes were evaluated by measuring the thickness of the functional zones of the adrenal cortex. Results: Decreased plasma CORT concentrations, as well as decreased adrenal CORT, desoxyCORT and 11-dehydroCORT concentrations, were observed in high- but not in low-anxiety phenotypes. These decreases were associated with increases in AI. PSS led to a significant decrease in the thickness of the zona fasciculata and an increase in the thickness of the zona intermedia. The increase in the thickness of the zona intermedia was more pronounced in low-anxiety than in high-anxiety rats. A decrease in the adrenal capsule thickness was observed only in low-anxiety rats. The nucleus diameter of cells in the zona fasciculata of high-anxiety rats was significantly smaller than that of control or low-anxiety rats. Conclusion: Phenotype-associated changes in adrenal function and histomorphology were observed in a rat model of complex post-traumatic stress disorder.

Age-Related Individual Behavioural Characteristics of Adult Wistar Rats.

Simple SummaryRats are considered adults from 2 to 5 months. During this period, they are used for experimentation in physiology and pharmacology. Adult rats, depending on their age, can be in a different physiological state, which can influence the results of experiments carried out on them. Despite this, age-related changes in adult rats have not yet been examined. Our results showed that as male and female rats progressed from 2 to 5 months of age there was a decrease in the level of motor and exploratory activities, and an increase in the level of anxiety-like behaviour. Age-related changes were dependent upon initial individual characteristics of behaviour. For example, animals that demonstrated high motor activity at 2 months become significantly less active by 5 months, and animals that showed a low level of anxiety at 2 months become more anxious by 5 months. Low-activity and high-anxiety rats did not show any significant age-related changes from 2 to 5 months of age. The results of this work should be taken into account when choosing the age of rats for conducting behavioural experiments.The aim of this work was to study age-related changes in the behaviour of adult Wistar rats using the open field (OF) and elevated plus maze (EPM) tests. Behavioural changes related to motor activity and anxiety were of particular interest. Results showed that as male and female rats progressed from 2 to 5 months of age, there was a decrease in the level of motor and exploratory activities and an increase in their level of anxiety. Age-related changes were dependent upon initial individual characteristics of behaviour. For example, animals that demonstrated high motor activity at 2 months become significantly less active by 5 months, and animals that showed a low level of anxiety at 2 months become more anxious by 5 months. Low-activity and high-anxiety rats did not show any significant age-related changes in OF and EPM tests from 2 to 5 months of age, except for a decrease in the number of rearings in the EPM. Thus, the behaviour of the same adult rat at 2 and 5 months of age is significantly different, which may lead to differences in the experimental results of physiological and pharmacological studies using adult animals of different ages.

Differential effects of early exposure to alcohol on alcohol preference and blood alcohol levels in low- and high-anxious rats.

Emotional disturbances emerge following alcohol withdrawal. The anxiolytic effect of alcohol may be one important motivation for its consumption in conditions where alcohol intake is anxiety reducing. Besides, early alcohol experience will predict future alcohol-related problems at some point in their lives. Rats classified according to their anxiety-like behavior phenotype show a higher preference for alcohol. Yet, despite preclinical studies have identified the behavioral and physiological effects of alcohol withdrawal, little has been shown on relapse to alcohol after a period of abstinence following intermittent long-term alcohol consumption in low-(LA) and high-anxiety (HA) phenotype rats. This study attempts to assess the role of emotional reactivity of early-aged LA and HA rats on later alcohol preference, through an operant response procedure. For this, a sweetened liquid fading procedure associated with a long-term and intermittent voluntary alcohol drinking was used, with the animals being tested on different withdrawal periods. Alcohol withdrawal symptoms were recorded, and blood alcohol levels were assessed at two intervals to examine the influence of anxiety phenotype on blood alcohol clearance. Data from HA control and HA withdrawn rats show that emotionality selectively influences alcohol preference. Blood alcohol decay was faster in HA than in LA alcohol pretreated rats. HA rats showed an ascending curve of alcohol intake, exhibiting a high level of alcohol drinking on withdrawal and withdrawal symptoms. Moreover, HA alcohol experienced rats show a high frequency of lever-pressing response during extinction, associated with a facilitation of bar-pressing recovery trials, an indication of alcohol-seeking behavior.

Effects of chronic stress on reinstatement of palatable food seeking: Sex differences and relationship to trait anxiety

Previous research in our lab has established a causal role for chronic stress exposure in subsequent increases in relapse-like behaviors in male rats with a history of palatable food self-administration. Given that many of the neurobehavioral consequences of stress are sex dependent, we aimed to determine whether sex differences exist with regard to the effects of chronic stress on relapse. Additionally, because high trait anxiety confers vulnerability to stress-related disorders, we examined whether individual differences in trait anxiety were related to differences in relapse-like behavior after chronic stress exposure. Following elevated plus maze testing for classification into high- or low-anxiety phenotypes, male and female rats responded for highly palatable food pellets. During subsequent extinction training, stress was manipulated (0 or 90 min restraint/day for 7 days). Rats were then tested for cue- and pellet priming-induced reinstatement of palatable food seeking. Results showed that female rats displayed higher levels of responding during cue-induced reinstatement tests compared to males, and that a history of chronic stress caused an attenuation of cue-induced reinstatement in female, but not male, rats. Regarding pellet priming-induced reinstatement, there was a three-way interaction such that neither stress history nor anxiety phenotype was related to reinstatement in females, but a history of stress in males caused increased and decreased responding in low- and high-anxiety rats, respectively. These results suggest that biological sex and trait anxiety level may help to explain differences in vulnerability to relapse among individuals exposed to chronic stress. Such information may be useful in designing more personalized and effective treatments for obesity and eating disorders.

5-HT1A and α2 adrenergic receptor levels are associated with high anxiety-like patterns and impulsivity in selectively bred Long Evans rats

Impulsivity and anxiety are psychological traits involved in many aspects of the drug addiction cycle. However, few preclinical models exist for examining both impulsive and anxiety patterns. In the current study, we investigated whether 6th generation rats selectively bred for high anxiety (HAn)-like behavior would display amphetamine (AMPH) hyperactivity. In the same generational line, we also determined if HAn animals would display impulsivity in an operant task. Filial 5 male Long Evans rats phenotyped as HAn and low anxiety (LAn) were tested on the elevated plus maze (EPM) and in locomotor chambers following a low dose of AMPH (0.5 mg/kg, IP). Next, a separate group of F5 animals was exposed to a differential reinforcement of low rate of responding (DRL: 30 s) operant schedule to assess impulsivity. Postmortem, 5-HT1A and α2 adrenergic receptor protein levels were measured in the medial prefrontal cortex (mPFC), nucleus accumbens (NAc) core and shell, and α2 adrenergic counts were assessed in the locus coeruleus (LC), and the paraventricular nucleus (PVN) of the hypothalamus. F5 outbred HAn rats had decreased percent open arm time and entries on the EPM and elevated AMPH-induced locomotion. In the DRL, HAn rats displayed an impulsive profile, they attained fewer total rewards, had more inter-response times, and showed greater burst ratios. We found that HAn rats had a higher number of 5-HT1A receptor immunostained cells in the mPFC but were not different than LAn in NAc core or shell. By contrast, levels of the α2 adrenergic receptor protein were no different in the mPFC while HAn rats had greater levels in the LC and lower levels in the PVN. Overall, these data further validate our outbred trait anxiety rats: HAn males show anxiety-like behavior, AMPH hypersensitivity, greater impulsivity, and varying levels of limbic and midbrain 5-HT1A and α2 adrenergic receptor proteins.

Endocrine, metabolic, and endocannabinoid correlates of obesity in rats exhibiting high anxiety-related behaviors

Abstract Anxiety disorders are major risk factors for obesity. However, the mechanisms accounting for this susceptibility remain unclear. Animal models have proved to be useful tools for understanding the role of emotional functioning in the development and maintenance of metabolic alterations implicated in obesity. Here we sought to determine the predictive value of behavioral indices of anxiety for hormonal and metabolic imbalances in rats. Adult Lewis rats were screened on the elevated plus maze (EPM). K-means clustering was used to divide the rats into two groups based on their anxiety index in the EPM: low (LA) and high anxiety (HA) rats. This proxy of anxiety combines individual EPM parameters and accepted ratios into a single score. Four weeks later, we measured markers of endocrine and metabolic function. We found that relative LA rats, the HA rats exhibited reduced latencies to exit a modified light-dark conflict test. Our results show that the HA rats displayed increased corticosterone levels when compared to LA rats. Furthermore, the HA rats weighted more and exhibited an enhanced glycemic response to exogenously administered glucose during the glucose tolerance test, indicating glucose intolerance. Notably, when compared to LA rats, the HA rats showed higher circulating levels of the endogenous cannabinoid, 2-arachidonoyl-sn-glycerol (2-AG). Together, these data indicate that patterns of emotional reactivity associated with anxiety may share common pathological pathways with metabolic complications implicated in obesity. Uncovering metabolic risk factors for anxiety disorders have the potential to strongly impact how we treat mental illnesses. Objective To test the hypothesis that indices of anxiety in the elevated plus maze have predictive value for alterations implicated in obesity, which include heightened emotional reactivity, and dysregulated corticosterone, glucose metabolism, and circulating endocannabinoid levels.

ВЛИЯНИЕ БЛОКАДЫ ГАМКА-РЕЦЕПТОРОВ НА ИЗМЕНЕНИЯ ОРИЕНТИРОВОЧНО-ИССЛЕДОВАТЕЛЬСКОЙ АКТИВНОСТИ И ТРЕВОЖНОСТИ, ВЫЗВАННЫХ КОРТИКОЛИБЕРИНОМ

The effect of the GABAA-receptor blockade with picrotoxin on the change in anxiety level under stress conditions induced by CRF was studied. In the T-maze test from the population of Wistar rats were selected animals with an active strategy of adaptive behavior, which in the elevated plus maze were divided into three groups: low anxiety (LA), high anxiety (HA) and mean anxiety (MA). Picrotoxin and CRF were injected intracerebroventricularly (ICV). The LA rats reacted to ICV administration of CRF with a decrease in the exploration activity, while the activity of the HA rats after the CRF injection was increased. The GABAA-receptor blockade with picrotoxin increased the exploration activity in HA and LA of rats, while simultaneously reducing the level of anxiety. After injection of CRF against of picrotoxin administration, the HA and LA rats demonstrated the decrease of exploration activity, and the level of anxiety increased only in LA rats. In animals from the MA group, the change in the behavioral response was observed only after the CRF and picrotoxin administration: the level of anxiety decreased, and the exploration activity increased. It is concluded that the vector of anxiety changes caused by CRF against the decrease in the activity of the stress-limiting GABA-ergic system depends on the initial psycho-emotional status of the animal.

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.

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.

Effects of Blockade of D1 and D2 Receptors in the Basolateral Amygdala on the Behavior of Rats with High and Low Levels of Anxiety and Fear

The effects of local bilateral administration of D1 or D2 receptor antagonists into the basolateral nucleus of the amygdala on anxiety behavior and the acquisition and extinction of conditioned reflex fear were studied in rats. Testing in the elevated plus maze identified groups of rats with high and low anxiety levels and assessment of the freezing duration after acquisition of conditioned reflex fear identified groups with low and high levels of freezing. Administration of a D1 receptor antagonist (SCH 23390, 1 μg/0.5 μl) decreased the signs of conditioned reflex fear in low-freezing rats in response to sound, accelerated its extinction, and degraded repeat learning in all animals. Administration of a D2 receptor antagonist (raclopride, 1 μg/0.5 μl) accelerated extinction of conditioned reflex fear to the experimental context in all rats and had weak anxiolytic actions in high-anxiety rats in the elevated plus maze. These data provide evidence for an important role for D1 receptors in the acquisition, manifestation, and extinction of conditioned reflex fear in response to the stimulus, while D2 receptors have an important role in the occurrence of anxiety states and fear in response to the context. Differences in the sensitivities to administration of dopamine receptor antagonists into the amygdala were seen in animals with different levels of anxiety and fear, which may be linked with differences in dopaminergic transmission.

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.

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.

Effects of oxytocin on background anxiety in rats with high or low baseline startle.

Oxytocin has antianxiety properties in humans and rodents. However, the antianxiety effects have been variable. To reduce variability and to strengthen the antianxiety effect of oxytocin in fear-potentiated startle, two experiments were performed. First, different amounts of light-shock pairings were given to determine the optimal levels of cue-specific fear conditioning and non-predictable startle (background anxiety). Second, the antianxiety effects of oxytocin were examined in rats with high and low pre-fear conditioning baseline startle to determine if oxytocin differentially affects high and low trait anxiety rats. Baseline pre-fear conditioning startle responses were first measured. Rats then received 1, 5, or 10 light-shock pairings. Fear-potentiated startle was then tested with two trial types: light-cued startle and non-cued startle trials. In the second experiment, rats fear conditioned with 10 light-shock pairings were administered either saline or oxytocin before a fear-potentiated startle test. Rats were categorized as low or high startlers by their pre-fear conditioning startle amplitude. Ten shock pairings produced the largest non-cued startle responses (background anxiety), without increasing cue-specific fear-potentiated startle compared to one and five light-shock pairings. Cue-specific fear-potentiated startle was unaffected by oxytocin. Oxytocin reduced background anxiety only in rats with low pre-fear startle responses. Oxytocin has population selective antianxiety effects on non-cued unpredictable threat, but only in rats with low pre-fear baseline startle responses. The low startle responses are reminiscent of humans with low startle responses and high trait anxiety.

Differential Effect of Unilateral Amygdalar GABA<sub>A</sub> Receptor Agonist Injection on Low- and High-Anxiety Rats

The influence of γ-aminobutyric type-A (GABAA) receptors agonist (muscimol hydrobromide, 0.1 μg/0.5 μl) injections into the right or left basolateral amygdala (BLA) on the behavior of high-anxiety (HA) and low-anxiety (LA) rats subjected to the elevated plus-maze (EPM) test was investigated. Anxiolytic-like effects (increase of open-arm entries and open-arm time) was revealed only after administration of muscimol into the left (but not right) amygdala of HA animals. No effect was observed upon administration of muscimol to LA rats. These findings suggest an important role in anxiety regulation of the amygdalar GABA levels, and the assumed GABA hemispheric lateralization.

Behavior of High- and Low-Anxiety Rats on the Background of Modulation of GABAergic Transmission in the Basolateral Nucleus of the Amygdala

The effects of local administration into the right or left basolateral amygdalar nucleus of a GABAA receptor agonist (muscimol hydrobromide, 0.1 μg/0.5 μl) or antagonist (bicuculline methiodide, 0.2 μg/0.5 μl) on behavior in the elevated plus maze were studied in high- and low-anxiety rats. High- and low-anxiety rats had different sensitivities to these substances. Administration of muscimol had anxiolytic actions only in high-anxiety rats, increasing the duration of excursions into the open arms of the maze. Administration of bicuculline led signs of aggressivity in all rats, with increases in motor and exploratory activity in high-anxiety rats and increases in the duration of excursions into the open arms of the maze and emotional tension in low-anxiety animals. More effective changes in behavior were seen with administration of muscimol into the left amygdala of high-anxiety rats and with bicuculline into the right amygdala of low-anxiety rats. These results provide evidence of the existence of individual-typological and interhemisphere differences in the functioning of the GABAergic system of the amygdala.