Long-term early life adverse experience impairs responsiveness to exteroceptive stimuli in adult rats.

It has been shown that early life traumatic events strongly alter the physiology and behavior in adult rats. In the present study, the effect of postnatal stressor on the spontaneous behavior of adult male rats was evaluated. A method of positive habituation based on a detailed analysis of behavioral patterns and attention of animals to a stimulus object was used. Twenty-four dams and twenty-four of their male progeny were used. Pups were divided into three groups (n = 8): controls (C); maternal social stressor (S); maternal social and physical stressors (SW). Animals (postnatal day 70-80) were individually placed in the open field arena in two habituation sessions with a 24-h delay between them (Test day 1 and Test day 2). Before the start of third session (Test day 3) a solid object was fixed in the center of the arena. Each test lasted 10 min. Our results showed the habituation effect in both stressed-groups. Although there were no significant differences in the number of investigations of the novel object among all tested groups, stress-exposed rats spent less time investigating the object. In conclusion, our findings indicate that long-term neonatal stress may impair an animal's ability to sustain attention to stimuli.

Can Anxiety Tested in the Elevated Plus-maze Be Related to Nociception Sensitivity in Adult Male Rats?

Methamphetamine (MA) is one of the most addictive psychostimulant drugs with a high potential for abuse. Our previous studies demonstrated that MA administered to pregnant rats increases pain sensitivity and anxiety in their adult offspring and makes them more sensitive to acute administration of the same drug in adulthood. Because individuals can differ considerably in terms of behaviour and physiology, such as rats that do not belong in some characteristics (e.g. anxiety) to average, can be described as low-responders or high-responders, are then more or less sensitive to pain. Therefore, prenatally MA-exposed adult male rats treated in adulthood with a single dose of MA (1 mg/ml/kg) or saline (1 ml/kg) were tested in the present study. We examined the effect of acute MA treatment on: (1) the anxiety in the Elevated plus-maze (EPM) test and memory in EPM re-test; (2) nociception sensitivity in the Plantar test; (3) the correlation between the anxiety, memory and the nociception. Our results demonstrate that: (1) MA has an anxiogenic effect on animals prenatally exposed to the same drug in the EPM; (2) all the differences induced by acute MA treatment disappeared within the time of 48 hours; (3) there was no effect of MA on nociception per se, but MA induced higher anxiety in individuals less sensitive to pain than in animals more sensitive to pain. In conclusion, the present study demonstrates unique data showing association between anxiety and nociceptive sensitivity of prenatally MA-exposed rats that is induced by acute drug administration.

Do the effects of prenatal exposure and acute treatment of methamphetamine on anxiety vary depending on the animal model used?

The aim of the present study was an evaluation of prenatal exposure to acute methamphetamine (MA) treatment on manifestations of anxiety. Anxiety was evaluated in adult animals in three different experimental models: the Elevated plus-maze (EPM), Social interaction test (SIT) and Ultrasound vocalization (USV). Female rats were administered saline (S) or MA (5 mg/kg) daily throughout their entire gestation period. The male progeny, in adulthood, were administered with challenge dose of S or MA (1 mg/kg) prior to evaluation of anxiety. The study demonstrated that prenatal MA exposure increased the anxiogenic effect on evaluated behaviour patterns in the USV model and to a lesser degree in the EPM model. In addition, the acute MA challenge in adulthood decreased the time spent during social interaction suggesting an anxiogenic effect in the SIT model as well. On the other hand, some of the evaluated parameters (e.g. the number of head-dipping in the EPM and number of dropped boluses in the SIT) also suggest MA-induced anxiolytic effects. Sensitization to a MA challenge was apparent in several parameters of the EPM (e.g. increased number of entries to the closed arms, increased stretched attend postures and increased approach-avoid conflicts) and SIT (total social interaction and following). The present data demonstrate that prenatal MA exposure and adult challenge of the same drug have diverse effects on animal behaviour that depends on the type of anxiety model used.

Effects of psychostimulants on social interaction in adult male rats.

Psychostimulants are known to have a huge impact on different forms of social behaviour. The aim of the present study was to compare the effects of three different psychostimulants [amphetamine, cocaine and 3,4 methylenedimethoxyamphetamine (MDMA)] on social interaction (SI) in adult male rats. The SI test was performed in a familiar arena and under low-stress environmental conditions. Experimental animals received amphetamine (0.5, 1.0, 1.5 mg/kg), cocaine (0.5, 1.0, 1.5, 2.5, 5.0, 10.0 mg/kg) or MDMA (2.5, 5.0, 10 mg/kg) and control animals received saline (1 ml/kg) 45 min before the SI test. Time spent in SI (individual patterns of social behaviour) and nonsocial activities (locomotion and rearing) were video recorded and then analysed offline, with the following results: (a) all doses of amphetamine decreased SI. Specifically, all doses of amphetamine decreased mutual sniffing, and the higher doses also decreased allo-grooming and following behaviours. (b) The higher doses of cocaine decreased SI, especially mutual sniffing, allo-grooming and climbing over. Cocaine at the dose of 5.0 mg/kg increased genital investigation compared with lower doses. (c) All doses of MDMA decreased mutual sniffing and climbing over; the two higher doses decreased allo-grooming behaviour, and only the highest dose decreased following. The two higher doses of amphetamine and all the doses of MDMA increased locomotion and rearing; cocaine did not affect locomotion, but increased rearing at higher doses. In conclusion, the results confirm the well-known finding that psychostimulants suppress SI, but also show novel differences in the effects of psychostimulants on specific patterns of SI.

An isolated epileptic seizure elicits learning impairment which could be prevented by melatonin.

We tested the relation between a single short tonic-clonic seizure elicited by flurothyl vapors and changes of learning in Morris water maze (MWM) in Wistar rats. Oxidative stress usually accompanies seizures. Large melatonin doses were applied immediately before and after seizures to test consequences on learning impairment. One hour of hypobaric hypoxia (8000 m) three days prior to the seizure served as an activator of intrinsic antioxidant systems. Learning in MWM (7 days) started 24 h after seizures. Following seizures, latencies in MWM were longer than in controls and were shortened by hypoxia and preventive melatonin application. Melatonin was also applied before hypoxia to influence free radical (FR) production and intrinsic antioxidant activation. Some behavioral characteristics were changed and preconditioning effect of hypoxia was reduced. Melatonin after seizure (150 s and 6 h) had negligible effect. Results allow us to hypothesize about the role of FR and the beneficial effect of melatonin on the behavioral consequences of seizures.

Modulations of behavioral consequences of minor cortical ischemic lesion by application of free radicals scavengers.

Functional and morphological consequences of ischemic lesions are partially related to the production of reactive oxygen species (ROS). The aim of the study was to create a unilateral photothrombic lesion with minimal morphological changes and minor sensorimotor and cognitive deficits and also to test whether the application of ROS scavengers after the end of induction of ischemia had improved the functional outcome. Adult Wistar male rats were randomly divided into five groups: naive control, sham operated animals, animals with induced ischemia, and two groups of animals with induced ischemia and subsequent ROS scavenger application -melatonin or tempol. The group subjected to ischemia showed a significant decline in performance in sensorimotor tests and the Morris water maze (MWM) test, compared to control animals. Tempol (50 mg/kg, i.p.) did not improve sensorimotor function and did not change spatial learning. Melatonin (100 mg/kg, i.p.), on the contrary, resulted in a significant improvement in animals' performances. All the ischemia subjected animals had increased speed of swimming in the MWM test, compared to the control group. Our findings showed that subsequent application of ROS scavengers improve ischemia outcomes, with melatonin being more potent. Conversely, neither melatonin, nor tempol decreased swimming speed cased by ischemia.

Sex differences in social interaction of methamphetamine-treated rats.

Our previous study showed that single injection of methamphetamine decreases social interaction (SI) in a dose-dependent manner that was further affected by stressful environment conditions. The aim of this study was to examine the effect of methamphetamine and its interaction with gonadal hormones on SI. Adult male and female rats were gonadectomized and assigned to testosterone-treated and oil-treated groups in male rats and estradiol-treated and oil-treated groups in female rats, respectively. Hormones were administered 30 min before each habituation in the open field. All four hormonal groups were further divided to control (without injection), saline (1 ml/kg saline injection), and methamphetamine (1 mg/kg) groups. Injections were applied 30 min before the SI test. The total duration and the total number of SI and nonsocial behavioral patterns were assessed. This study showed that an acute methamphetamine administration in a dose of 1 mg/kg decreased different types of SI in both sexes. In contrast, the same dose of methamphetamine increased locomotion and rearing behavior in male and female rats. The frequency and/or duration of SI (especially mutual sniffing and allogrooming) was lower in adult female rats relative to gonadectomized male rats, but locomotion was increased in female relative to male rats regardless of the presence or absence of gonadal hormones. In conclusion, this study is novel especially because it examines SI in both sexes in relation to the presence or absence of gonadal hormones.

Does prenatal methamphetamine exposure affect the drug-seeking behavior of adult male rats?

Methamphetamine (MA) is one of the most frequently used illicit drugs worldwide and also one of the most common drugs abused by pregnant women. Repeated administration of psychostimulants induces behavioral sensitization in response to treatment of the same or related drugs in rodents. The effect of prenatal MA exposure on sensitivity to drugs in adulthood is not yet fully determined. Because our most recent studies demonstrated that prenatal MA (5mg/kg) exposure makes adult rats more sensitive to acute injection of the same drug, we were interested whether the increased sensitivity corresponds with the increased drug-seeking behavior. The aim of the present study was to examine the effect of prenatal MA exposure on drug-seeking behavior of adult male rats tested in the conditioned place preference (CPP). The following psychostimulant drugs were used as a challenge in adulthood: MA (5mg/kg), amphetamine (5mg/kg) and cocaine (10mg/kg). All psychostimulant drugs induced increased drug-seeking behavior in adult male rats. However, while MA and amphetamine-induced increase in drug-seeking behavior did not differ based on the prenatal drug exposure, prenatally MA-exposed rats displayed tolerance effect to cocaine in adulthood. In addition, prenatally MA-exposed rats had decreased weight gain after administration of MA or amphetamine, while the weight of prenatally MA-exposed rats stayed unchanged after cocaine administration. Defecation was increased by all the drugs (MA, amphetamine and cocaine), while only amphetamine increased the tail temperature. In conclusion, our results did not confirm our hypothesis that prenatal MA exposure increases drug-seeking behavior in adulthood in the CPP test.

The effect of methamphetamine on social interaction of adult male rats.

The effect of psychostimulants on social behavior still remains unclear. Therefore the aim of the present study was to assess the effect of low doses of methamphetamine (MA) on social interaction (SI) in adult male rats. Rats were tested in three environmental conditions: (1) dimly lit, familiar environment, (2) dimly lit, unfamiliar environment and (3) intensely lit, unfamiliar environment considered to be low, middle and high stress, respectively. In each condition different set of animals was used. Rats were always divided into five groups. Control (without injection), saline (with 1 ml/kg saline injection) and three MA groups (doses: 0.5, 1 and 1.5mg/kg). Injections were applied 30 min prior to testing. Always a pair of unfamiliar rats of the same treatment group was tested. Their behavior was video recorded for 5 min in an open field. Times spent by SI (following, climbing, genital investigation, etc.) and non-social behavior (locomotion, rearing) were analyzed using a two-way ANOVA (drug treatment x stress condition). Our data demonstrate that all doses of MA, reduced SI. In addition, the unfamiliarity of the arena increased exploratory behavior (locomotion and rearing) in all treatment groups, while the SI was affected by the environmental condition only in controls or saline-treated rats, but not in MA-treated groups. In conclusion, our data demonstrate that MA administration impairs SI in dose- and stress condition-specific manner, however, some of our results may be due to increase locomotion and rearing induced by MA.

Effects of 2-chloroadenosine on cortical epileptic afterdischarges in immature rats.

Adenosine may represent an endogenous anticonvulsant in the brain. This study focused on the possible anticonvulsant action of an adenosine agonist, 2-chloroadenosine, against cortical epileptic afterdischarges (ADs) in immature rats. Three age groups of rat pups with implanted electrodes were studied: 12-, 18- and 25-days-old. The compound, 2-chloroadenosine, was injected after the first successful stimulation at doses of 1, 4 or 10 mg/kg intraperitoneally, and stimulation at the same intensity was repeated three more times. Movements directly elicited by stimulation, as well as clonic seizures accompanying electroencephalography (EEG) ADs, were markedly suppressed in only the 18-day-old animals. The effects in the 12- and especially the 25-day-old rats were moderate. The duration of the ADs decreased in all three age groups with 2-chloroadenosine treatment, and the shortest AD duration was seen in the treated, 12-day-old rats. The AD suppression also lasted longer in this age group than it did in the older animals. After a brief suppression of the second AD, the treated, 25-day-old group exhibited a significant AD rebound during the third and fourth stimulations. Taken together, our data show that 2-chloroadenosine exhibits an anticonvulsant effect that is dose- and age-dependent.

Effect of methamphetamine exposure and cross-fostering on cognitive function in adult male rats.

The aim of our study was to examine the effect of prenatal methamphetamine (MA) exposure and cross-fostering on cognitive functions of adult male rats tested in Morris water maze (MWM). Rat mothers were exposed daily to injection of MA (5mg/kg) or saline for 9 weeks: prior to impregnation, throughout gestation and lactation periods. Females without any injections were used as an absolute control. On postnatal day 1, pups were cross-fostered so that each mother raised 4 pups of her own and 8 pups from the mothers with the other two treatments. Four types of tests were used: (1) Place navigation test (Learning), (2) Probe test (Probe), (3) Retention memory test (Memory) and (4) Visible platform task. Our results demonstrate that the prenatal exposure to MA does not impact learning and memory, while postnatal exposure to MA shows impairments in cognition. In the test of learning, all animals fostered to MA-treated dams had longer latencies, bigger search error and used lower spatial strategies than the animals fostered to control or saline-treated mother, regardless of prenatal exposure. Regardless of postnatal exposure, the animals prenatally exposed to saline swam faster in all the tests than the animals prenatally exposed to MA and controls, respectively. This study indicates that postnatal but not prenatal exposure to MA affects learning in adult male rats. However, it is still not clear whether these impairments are due to a direct effect of MA on neuronal structure or due to an indirect effect of MA mediated by impaired maternal care.

Effect of methamphetamine exposure and cross-fostering on sensorimotor development of male and female rat pups.

The present study tested the hypothesis that cross-fostering influences the development of rat pups. Mothers were exposed daily to injection of methamphetamine (M) (5 mg/kg) or saline for 9 weeks: 3 weeks prior to impregnation, throughout gestation and lactation periods. Control females animals without any injections were used. On postnatal day (PD) 1, pups were cross-fostered so that each mother received four pups of her own and eight pups from the mothers with the other two treatments. Offspring were tested for sensorimotor development in preweaning period by using tests of: negative geotaxis, tail pull, righting reflexes, rotarod and bar-holding. Further, the pups were weighed daily. Our results showed that birth weight in prenatally M-exposed pups was lower than in control or saline-exposed pups. Prenatally M-exposed pups gained less weight than control or saline-exposed pups regardless of postnatal treatment and sex. Further, our data demonstrated that prenatal and postnatal M exposure impairs sensorimotor functions in most of the tests. On the other hand, the negative effect of prenatal M exposure was partially suppressed in prenatally M-exposed pups by cross-fostering to control dams. Our hypothesis that cross-fostering may affect postnatal development of pups was confirmed.

Cross-fostering effect on postnatal development of rat pups exposed to methamphetamine during gestation and preweaning periods.

There are studies showing that drug abuse during pregnancy may have a long-term effect on progeny of drug-abusing mothers. Our previous work demonstrated that prenatal and/or postnatal methamphetamine injections impair maternal behavior. The purpose of the present study was to assess the effect of prenatal methamphetamine or stress exposure and postnatal breeding on postnatal development of rat pups. Female rats were injected with methamphetamine (5 mg/kg daily) or physiological saline prior, during and after gestation. Absolute controls did not receive any injections. On postnatal day 1, pups were cross-fostered so that each mother received some of her own and some of the pups from the mothers with the other two treatments. Pups were weighted daily for the entire lactation period. Postural motor reaction development was examined daily by righting reflex between postnatal day 1 and 12. On postnatal day 15 homing test examining pups' nest-seeking behavior was performed. On postnatal day 23 rotarod and bar-holding tests were used to investigate sensorimotor coordination of pups. We demonstrated that prenatal methamphetamine exposure impairs performance of sensorimotor tests (righting reflex on surface and rotarod test). Moreover, the effect of methamphetamine as well as the effect of prenatal stress induced by saline injections was affected by postnatal breeding conditions in sensorimotor tests as well as in the test of homing. Our results support the hypothesis that the variation in rat maternal care could serve as a mechanism for a nongenomic behavioral mode of transmission of traits.

Early stage of adjuvant arthritis alters behavioral responses in male but not female rats.

Anxiety and depression commonly occur in the pathology of rheumatic diseases. Little is known about how inflammatory disease in its early stage, before any clinical manifestation, may affect general activity. The aim of this study was to compare the anxiety-like behaviour in the early stage of adjuvant arthritis (AA), and the paw edema, and corticosterone (CORT) levels in the developed stage of AA among male and female Long Evans rats. The behavioural activity was evaluated by elevated plus maze tests. These revealed significantly reduced number of entries into the open arm of the maze in arthritic males compared to controls or to females 4 days after AA induction. Arthrihtic and control females did not differ. The number of entries into the closed arm of the maze was the same across the genders and studied intervals. Time spent in the open arm was significantly lower in arthritic males against controls or arthitic females. Time spent in the closed arm showed inverse picture to the time spent in the open arm. Hind paw swelling measured on day 23 of AA was the same in males and females, as was the elevation of CORT levels in plasma. Male rats showed anxiety-like behaviour on day 4 of AA, while female rats did not show any change, indicating different brain sensitivity to early inflammation among the genders.

Effect of methamphetamine exposure during prenatal and preweaning periods lasts for generations in rats.

Our previous studies demonstrated that methamphetamine (MA) administration during gestation and/or lactation affects maternal behavior in rats and that birth weight and sensory-motor coordination of their pups are also influenced. The present study tested the hypothesis that the effect of MA induces long-term changes affecting second generation of rats that were not exposed to the drug. Adult females exposed during prenatal and preweaning periods to 5 mg/kg MA daily, were examined for regularity of estrous cycle and mated with stimulus, unexposed males. Dams (nontreated absolute control, saline- and MA-exposed) were observed with their pups in two tests of maternal behavior (observational and retrieval tests). Their pups were further tested throughout the preweaning period to examine their development. Our data demonstrate that MA-exposed mothers displayed more nursing, were more often in the nest and in contact with their pups, and were faster in retrieving their pups than saline-exposed and/or control mothers. There were no differences in litter characteristics, birth weight and weight gain of pups between groups. Interestingly, pups from mothers exposed to MA during prenatal and preweaning period had impaired sensory-motor coordination. They achieved righting reflex in mid-air later than both control groups. Additionally, they had more falls in rotarod and bar-holding tests than pups from both control and saline-exposed mothers. In homing performance, pups from MA- and saline-exposed dams learned slower to return to the home box than pups from control dams. Thus, the present study demonstrates that MA abused by mothers may affect two generations of their offspring.

Postnatal development of rat pups is altered by prenatal methamphetamine exposure.

There are studies showing that drug abuse during pregnancy may have impairing effect on progeny of drug-abusing mothers. Methamphetamine (MA) is one of the most common illicit drugs throughout the world. The purpose of the present study was to assess the effect of prenatal MA exposure on postnatal development of rat pups before the time of separation from their mothers. Female rats were injected with MA (5 mg/kg daily) for the duration of their pregnancy. Pups were then tested throughout the lactation period. They were weighed daily and the ano-genital distance was measured on postnatal day (PD) 1. Development of postural motor reaction was tested by righting reflex on surface between PD 1 and 12, and righting reflex in mid-air after PD 12 until successfully accomplished. On PD 15 homing test was examined as a test of pup acute learning. On PD 23 sensory-motor coordination was examined using the rotarod and bar-holding tests. Additionally, the markers of physical maturation, such as eye opening, testes descent in males and vaginal opening in females were also recorded. The birth weight in prenatally MA-exposed pups was lower than in controls or saline-exposed pups regardless of sex. There were no changes induced by prenatal MA exposure in weight gain or in sexual maturation. In righting reflexes, we demonstrated that pups exposed prenatally to MA were slower in righting reflex on surface and that they accomplished the test of righting reflex in mid-air later than controls or saline-exposed pups. The performance of homing test was not affected by prenatal drug exposure. The sensory-motor coordination was impaired in prenatally MA-exposed pups when testing in the rotarod test. Also, the number of falls in the bar-holding test was higher in MA-exposed pups than in controls. There were no sex differences in any measures. Thus, the present study demonstrated that prenatal MA exposure impairs development of postural motor movements of rat pups during the first 3 weeks after birth, while not affecting physical or sexual maturation.

Learning in the Place navigation task, not the New-learning task, is altered by prenatal methamphetamine exposure.

Adult rats (prenatally methamphetamine-exposed, saline-exposed and controls) were tested for learning in the Morris water maze. Rats were tested in 4 consecutive days using two different types of learning tests: "Place navigation test" (stable platform position) and "New-learning test" (platform position changed daily). Males exposed prenatally to methamphetamine were slower in the Place navigation learning test than were both the control and saline-exposed males. There were no differences in the New-learning test between groups.

Maternal behavior is impaired by methamphetamine administered during pre-mating, gestation and lactation.

Previous studies demonstrated that stimulant drugs, such as cocaine or amphetamine, administered during gestation or lactation may attenuate maternal behavior in rats. The effect of methamphetamine (MA), a drug whose usage has increased lately, on maternal behavior has not yet been investigated. The present study tested the effect of MA (5 mg/kg daily) administered prior to, during and after gestation on maternal behavior. Regularity of the estrous cycle, the incidence of impregnation, and the weight gain was compared between groups (control, saline- and MA-treated). Maternal behavior was examined using two tests: Observation test (without disturbance of the mother and pups) and Retrieval test (with short separation of pups from the mother). All tests were conducted prior to dosing each day. In the Observation test, MA decreased the blanket position of active nursing, while increasing passive nursing. There were no MA-induced changes in other maternal activities such as mother being in the nest, in contact with pups, or grooming pups. MA increased some non-maternal activities, such as drinking, eating, and sleeping, while decreasing stereotypic behavior (sniffing and rearing) when compared to controls. In the Retrieval test, MA-treated mothers were slower in retrieving the first pup, returning the first pup into the nest, and returning all pups into the nest. Interestingly, there were differences in maternal behavior also in saline-treated mothers relative to controls. Saline-treated mothers spent more time in the nest and groomed pups more than controls or MA-treated mothers. In conclusion, the present study demonstrates a novel finding that MA administered during pre-mating, gestational and lactational periods has a negative effect on maternal behavior toward pups.

Methamphetamine administration during gestation impairs maternal behavior.

Previous studies demonstrated that repeated drug exposure, such as opiates or cocaine, during the gestation period attenuates maternal behavior of rats; however, it is not known whether methamphetamine (MA), a drug whose usage has increased recently, negatively affects maternal behavior as well. Therefore, the present study tested the hypothesis that repeated subcutaneous administration of MA (5 mg/kg daily) throughout the entire gestation period alters maternal behavior. Dams (control, saline-, and MA-treated) were observed with their pups in two types of tests. In the observation test, 11 types of activities and three types of nursing positions of mothers were recorded 10 times during each 50-min session for the 22-day lactation period. A decrease in nursing and active maternal behavior was found in MA-treated mothers relative to control rats. In addition, stereotypic behavior such as rearing and sniffing was increased in MA- as well as in saline-treated mothers relative to controls. All mothers, regardless of the treatment, displayed significantly less maternal behavior and more nonmaternal activities as postpartum time progressed. In the retrieval test, mothers also were tested for pup retrieval from postpartum Days 1 through 12. MA-treated mothers were slower in retrieving the first pup, returning the first pup into the nest, and returning all pups into the nest relative to controls or saline-treated mothers. Interestingly, the latency to return all pups to the nest was longer in saline-treated mothers relative to controls. In conclusion, the present study demonstrates a novel finding that MA administered during the gestation period has a negative effect on maternal behavior.

Neuroendocrine or behavioral effects of acute or chronic emotional stress in Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats.

OBJECTIVE: Spontaneously hypertensive rats (SHR) selected from Wistar Kyoto (WKY) strain represent an animal model of human essential hypertension. This strain of rats is known by excessive neuroendocrine and cardiovascular responses under stress. The aim of the present study was: 1. To compare the reactivity of hypothalamic-pituitary-adrenocortical axis (HPA) to acute mild stress of handling between SHR and WKY rats, 2. to compare the behavioral activity of both strains under basal conditions and during chronic unpredictable emotional stress. METHODS: Seven to eight weeks old male SHR and WKY rats bred in the Physiological Institute, Academy of Sciences of the Czech Republic (Prague) were used. Acute stress was induced by 2-minute handling of the animals in their cage. Blood plasma was analyzed for ACTH and corticosterone (CORT) by specific radioimmunoassay. Chronic unpredictable stress lasted 20 days and consisted of random exposures to following interventions: Light on or off for 24 h, overcrowding i.e. pooling the rats from two cages into one (size 24 x 39 x 23 cm) for 24 h, isolation by placing a single rat into one cage for 24 h, new hierarchy by mixing 4 rats from two different cages for 24 h, limited access to food or water for 1 hour in one day between 3 and 6 p.m., inescapable foot shock (20 shocks, duration 5 s, intensity 10 mA, intershock interval 30 s), tilting the cages for 24 h. The sequence of individual stress exposures was the same in all rats. On day 6, 10 and 20, behavioral activity was measured using the elevated plus-maze in non-stressed control and stressed rats. The results were evaluated by non parametrical Kruskal-Wallis test followed by Mann-Whitney U-test. RESULTS: The two-minute handling resulted in a significantly higher activation of HPA in the SHR than in the WKY rats (plasma ACTH: 350 +/- 65 pg/ml for SHR vs. 97 +/- 17 pg/ml for WKY p<0.01; plasma corticosterone: 2.8 +/- 1.4 mg/100 ml for SHR vs. 0.7 +/- 0.06 mg/100 ml for WKY p<0.05). In WKY rats no activation of HPA was observed. Elevated plus-maze anxiety test showed inverse behavioral pattern between SHR and WKY rats. In the first test of anxiety the number of open arm entries (OAE) as well as total mobility expressed as total arm entries of the SHR was lower than of the WKY rats (p<0.01) without any difference between stressed and non-stressed animals in either strain. It was gradually increasing in stressed and non-stressed SHR in subsequent sessions markedly exceeding the activity of WKY rats (p<0.01). Stressed WKY rats showed less OAE and total mobility than their controls (p<0.01). CONCLUSIONS: Our results show enhanced neuroendocrine response to acute handling and enhanced anxiety in acute novelty stress in SHR comparing to WKY rats which suggests a common mechanisms for neuroendocrine and behavioral changes. These results further underline the lack of anxiety related behavior of SHR under chronic emotional stress.