Acquisition of DRL-20 behavior in male and female, Roman high- and low-avoidance rats

Female RLA rats were more efficient than male RLA, male and female RHA rats (in that order), in the acquisition of DRL-20 operant conditioning behavior. It was concluded that RHA rats were less capable of inhibiting nonrelevant activity in that test.     

Two distinctive apomorphine-induced phenotypes in the Roman high- and low-avoidance rats

Strain differences in spontaneous locomotor activity and the behavioral patterns induced by dopaminergic agonists in rodents can disclose differential genetic susceptibilities to dopaminergic dysfunction (i.e. vulnerability to psychosis). Psychogenetic selection of hypoemotional Roman High-Avoidance (RHA) and hyperemotional Low-Avoidance (RLA) rats leads to divergence in dopaminergic function as well. The present study was designed to characterize their spontaneous activity and their responses to apomorphine (0.067-3 mg/kg, s.c.) as compared to those of the standard Sprague-Dawley (SD) strain. The Roman strains displayed higher spontaneous activity than SD rats and RHA exhibited the higher response to novelty which agrees with a higher sensitivity to apomorphine in this strain. The biphasic effect induced by apomorphine (locomotor inhibition and yawning at low doses but stimulation of locomotion and stereotyped behavior at higher ones) was reproduced in the standard SD strain. Low doses were less effective inducing locomotor inhibition in RHA whereas these animals were much more sensitive to high dose-induced stereotyped behavior. In contrast, RLA was characterized as a high-yawning strain and low doses of apomorphine also induced a striking motor inhibition suggesting functional enhancement of dopamine receptors mediating these behaviors. The detailed and distinctive behavioral profiles described in this work suggest between strain differences both at the presynaptic and postsynaptic dopaminergic function and may serve as paths to better specify functional mechanisms in future studies of risk of developing dopaminergic dysfunctions.     

Context-dependent differences in grooming behavior among the NIH heterogeneous stock and the Roman high- and low-avoidance rats

Grooming occurs during/after stress and seems to accompany dearousal. Here, grooming was investigated under testing situations involving different levels of aversiveness, taking advantage of differences among three rat strains in fearfulness/anxiety. Inbred Roman High Avoidance (RHA-I) rats are less anxious/fearful than inbred Roman Low Avoidance (RLA-I). The outbred genetically heterogeneous stock of rats (NIH-HS), which resembles the RLA-I in many behavioral traits, was also studied. Adult male rats (RLA-I: n = 9, RHA-I: n = 10, NIH-HS: n = 12) were observed for 30 min in: a novel open-field, a novel hole-board and in the home-cage. They were also observed during two-way active avoidance training. Differences in grooming depended on test situation: (a) No differences were found in the home-cage. (b) While tested in a novel environment, RHA-I showed less grooming activity than the other rats. (c) After avoidance responses appeared, differences among the strains were opposite to the observed in novelty tests. Furthermore, results suggest that (i) grooming is mostly suppressed when assured aversive experience is under way; (ii) rostral grooming prevails when experience with aversive stimuli is unpredictable (novelty) or potential (avoidance training); (iii) body grooming increases for a period in novel environments. In general, our results support that grooming takes place during dearousal.     

Differential hormonal and physiological responses to stress in Roman high- and low-avoidance rats

In the present experiments, plasma concentrations of corticosterone, ACTH, prolactin, blood glucose, serum free fatty acids, core temperature and fecal boli were measured immediately following the exposure of Roman high- and low-avoidance rats (RHA/Verh, RLA/Verh) to various stressful situations. As compared to RLA/Verh rats, the RHA/Verh animals showed an attenuated emotional response in “non-self-menacing” (novel environment) situations. Differences between the 2 selected lines were not generally found in unstressed rats or in “high-stress” (ether, immobilisation, footshock) situations.     

Differential hippocampal neuron density between inbred Roman high- (low anxious) and low-avoidance (high anxious) rats

The inbred Roman low- (RLA-I) and high-avoidance (RHA-I) rats used in this study were initially selected and bred for extremely poor vs. rapid acquisition of active two-way avoidance behavior in the shuttle box. As a result of the selection for divergent avoidance acquisition, clear behavioral differences have been found between RHA and RLA rats in a variety of tasks related to anxiety and conflict. In rats of these two strains/lines previous brain studies have been performed, specifically in the striatum, the mesencephalic dopaminergic areas and the prefrontal cortex, as these brain areas are the classical ones for their critical role in sensitization and may play a role in the well-characterized anxiety response. In this study we analyzed, in RHA and RLA groups (N = 5 each), the density of NeuN neurons counterstained with toluidine blue in the cingulate cortex (subdivision 1) and the hippocampus (CA1, CA2 and CA3). A statistical difference was found in the density of neurons of CA1 and CA2 (p = 0.047 in both) and in the total density of the hippocampus (p = 0.009). Contrary to our expectations, significant strain differences for the density of neurons in the cingulate cortex were not found. The relationship between those differences in the hippocampus and the between-strain differences in anxiety and in learning processes depending on anxiety are discussed.     

Differential hippocampal neuron density between inbred Roman high- (low anxious) and low-avoidance (high anxious) rats

Nicotine, the addictive agent in cigarettes, reduces circulating estradiol-17β (E₂) and inhibits E₂-mediated intracellular signaling in hippocampus of female rats. In hippocampus, E₂-signaling regulates synaptic plasticity by phosphorylation of the N-methyl-d-aspartic acid receptor subunit NR2B and cyclic-AMP response element binding protein (pCREB). Therefore, we hypothesized that chronic nicotine exposure induces synaptic dysfunction in hippocampus of female rats. Female rats were exposed to nicotine or saline for 16 days followed by electrophysiological analysis of hippocampus. Briefly, population measurements of excitatory post-synaptic field potentials (fEPSPs) were recorded from stratum radiatum of the CA1 hippocampal slice subfield. A strict software-controlled protocol was used which recorded 30 min of baseline data (stimulation rate of 1/min), a paired-pulse stimulation sequence followed by tetanic stimulation, and 1 h of post-tetanus recording. EPSP amplitude and the initial EPSP slope were measured off-line. We then investigated by Western blot analysis the effects of nicotine on hippocampal estrogen receptor-beta (ER-β), NR2B and pCREB. The results demonstrated significantly decreased post-tetanic potentiation and paired-pulse facilitation at the 40, and 80 ms interval in nicotine-exposed rats compared to the saline group. Western blot analysis revealed that nicotine decreased protein levels of ER-β, NR2B, and pCREB. We also confirmed the role of E₂ in regulating NR2B and pCREB phosphorylation by performing Western blots in hippocapmal tissue obtained from E₂-treated ovariectomized rats. In conclusion, chronic nicotine exposure attenuates short-term synaptic plasticity, and the observed synaptic defects might be a consequence of loss of estradiol-17β-signaling. However, determining the exact molecular mechanisms of chronic nicotine exposure on synaptic plasticity specific to the female brain require further investigation.     

Tail pinch induced eating: psychogenetic comparison of Roman high- and low-avoidance rats

Consummatory behavior elicited by mild tail pinch in the presence of solid food was studied in male Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLA/Verh) rats. RLA/Verh rats, characterized as the more emotional of the two rat lines, exhibited significantly shorter latencies to bite and eat solid food during tail pinch than the RHA/Verh rats. Treatment with 6, 12 and 24 mg/kg sodium pentobarbital prior to tail pinch failed to increase the elicited eating in either rat line.     

Inbred Roman high- and low-avoidance rats: differences in anxiety, novelty-seeking, and shuttlebox behaviors.

In the present study, male inbred animals (from the 10th generation of an inbreeding program that has been carried out in parallel to that of the outbred Roman high- and low-avoidance rat lines), were compared for emotionality in different testing situations, exploratory behavior in the holeboard and two-way, active-avoidance acquisition. Compared to the inbred Roman high-avoidance (RHA-I/Verh) rats, inbred Roman low-avoidance (RLA-I-Verh) rats showed higher emotionality in the open field (reduced distance travelled and number of rearings, and increased self-grooming behavior), in the elevated plus-maze test (increased number of total and open-arm entries, reduced distance travelled in the open arms, and increased self-grooming behavior), and during the habituation period in the shuttle box (decreased number of crossings, increased self-grooming behavior and defecations). Results from the hyponeophagia test were not conclusive, probably due to the test-dependent hyperactivity shown by RHA-I/Verh rats. In the holeboard apparatus, RHA-I/Verh rats explored more than RLA-I/Verh rats, especially when novel objects were located beneath the holes. Finally, RHA-I/Verh animals rapidly acquired active, two-way (shuttlebox) avoidance, whereas RLA-I/Verh animals required four 50-trial sessions to achieve an assymptotic level of 30-40% avoidance. Thus, the behavioral patterns of the Roman inbred strains were very similar to those previously reported for the RHA/Verh outbred lines. Differences in locomotor activity, exploratory, and self-grooming behavior were actually greater between the inbred strains than between the outbred lines. Differences in defecation, however, although still significant, were not so pronounced as those noted previously at this laboratory with the outbred lines.     

Successive positive contrast in one-way avoidance behavior with Roman low-avoidance rats

The inbred Roman High- (RHA-I) and Roman Low-Avoidance (RLA-I) rats, psychogenetically selected for rapid (RHA-I) vs. extremely poor (RLA-I) acquisition of two-way active avoidance, exhibit a lower or a higher level of fearfulness, respectively, that can be observed in many laboratory anxiety models. The present study analyzed the performance of female RLA-I and RHA-I rats in a successive positive contrast situation induced during one-way avoidance learning. Three groups of RLA-I and three of RHA-I rats (1–30, 30–30 and 1–1 groups, the numbers stand for the time spent in the safe compartment during the first and second phase of training) were trained to avoid an electric foot-shock administered in a “danger” compartment, by running from this compartment to a “safe” one. Only RLA-I rats showed a significant positive contrast effect, in such a way that the reinforcement increase from the lower (1 s spent in safety) to the higher reward (30 s) led to a response enhancement, surpassing the performance of rats trained with the low (1–1 s) or the high (30–30 s) reward from the beginning of training. The results are discussed in the context of an opponent process theory based upon the interaction between the motivational strength of fear and the incentive value of relief taking place during one-way avoidance learning.     

One-way avoidance acquisition and cellular density in the basolateral amygdala: Strain differences in Roman high- and low-avoidance rats

The goal of the present experiment was to study the performance of inbred Roman high- (RHA-I) and low- (RLA-I) avoidance rats in one-way avoidance learning and to relate the behaviour of the animals to cellular density in the basolateral amygdala (BLA), a brain region related to fear and anxiety. Thus, females from both strains were exposed either to 30 s or 1 s in the safe place as a function of experimental condition, until they reached five consecutive avoidance responses. Thereafter, the rats were perfused, and their brains sectioned in 40 μm coronal sections, stained with cresyl violet. The area (percentage of field) corresponding to the BLA structures was quantified by computerized-assisted image analysis. The results indicated that RLA-I showed a significantly poorer acquisition of the one-way avoidance task than did RHA-I rats, but only when safe time was the shortest (1 s). In addition, the number of trials needed to reach the behavioural acquisition criterion was negatively correlated with BLA cellular density in RLA-I rats. These data suggest the possibility of relating behavioural and neuro-anatomical indexes, enabling exploration of the biological basis of fear/anxiety behaviours.     

One-way avoidance learning in female inbred Roman high- and low-avoidance rats: Effects of bilateral electrolytic central amygdala lesions

Female inbred Roman high- (RHA-I) and low- (RLA-I) avoidance rats show differences in one-way avoidance learning only when the task implies a highly aversive situation (1 s in the “non-shock”-associated safe compartment, as opposed to 30 s). These between-strain differences seem to depend on strain differences in emotionality, given that: (i) they are abolished by IP administration of the GABAergic anxiolytic diazepam (Torres et al. [32]) and (ii) avoidance responding appears to correlate with cellular density in the basolateral amygdala (Gómez et al. [9]). In the present study we further analyzed whether the implication of the amygdala in one-way avoidance depends on the experimental situation aversiveness (30 s vs. 1 s in safety). After bilateral electrolytic lesions (1 mA; 20 s) of the central amygdala (CeA), RHA-I and RLA-I rats were exposed to a danger compartment (where they received a warning signal – 88 dB tone – followed by a 1 mA electric foot-shock), and a safe compartment, where these stimuli were not presented. The number of trials needed to reach 5 consecutive avoidance responses was used as dependent variable. Sham lesioned RLA-I rats showed poorer performance than sham lesioned RHA-I rats only under the 1 s condition. The CeA lesion disrupted the avoidance response only in 1 s groups, abolishing the between-strain performance differences observed under this condition. These results indicate the implication of CeA in one-way avoidance performance, and suggest a reciprocal modulation of fear and reinforcement (i.e. fear relief) in this form of aversive learning.     

Microstructural analysis of chlordiazepoxide's effects on food preference behavior in roman high-, control- and low-avoidance rats

The effects of chlordiazepoxide (CDP) on feeding performance and on rearing activity were investigated using a food preference test in three Roman strains: Roman Low Avoidance (RLA), Roman Control Avoidance (RCA), and Roman High Avoidance (RHA). The aims of the study were to assess the responses of the three strains to a free choice of familiar and novel foods following a period of food deprivation, and to answer the important psychopharmacogenetic question of whether or not the strains would display differential responsivity to CDP challenges. No strain differences were discovered in relation to the latency to begin feeding, the response to familiar food, or the frequency of rearing in the 10 min food preference test. However, clear differences did emerge in relation to the animals' responses to novel foods. In contrast to the RCA and RHA lines, RLA rats sampled a greater variety of the available foodstuffs, and devoted a greater duration to the consumption of novel foods. Despite previous reports that under conditions of mild stress, RLA animals display greater emotionality, there was no evidence from the present study to show that RLA animals were more neophobic (a possible index of emotionality) than the other two Roman lines. Indeed, the evidence clearly suggests that RLA animals may under certain circumstances pay greater attention to novel cues. CDP treatments produced reductions in rearing, reductions in eating latency, and strongly potentiated the time devoted to feeding. Effects of CDP treatments on familiar and novel foods were dissociable. CDP at 10 mg/kg reliably increased the novel food feeding duration, an effect occurring mainly within the first 5 min of the test.(ABSTRACT TRUNCATED AT 250 WORDS)     

Differential gene expression between inbred Roman high- (RHA-I) and low- (RLA-I) avoidance rats

Microarray technology was used to explore differences in brain gene expression under basal conditions in two strains of psychogenetically selected rats which differ in anxiety/stress responses, the inbred Roman High-(RHA-I) and Roman Low-(RLA-I) Avoidance rats. Microarray analysis detected 14 up-regulated and 24 down-regulated genes in RLA-I vs. RHA-I rats functionally related to neurobiological processes. The differentially expressed genes CAMKK2, CRHBP, EPHX2, HOMER3, NDN, PRL and RPL6 were selected for microarray validation using qRT-PCR. EPHX2, CAMKK2 (both up-regulated in RLA-I vs. RHA-I rats) and HOMER3 (down-regulated in RLA-I vs. RHA-I rats) showed a similar tendency and fold-change both in microarray and RT-PCR analyses; PRL (up-regulated in RLA-I vs. RHA-I rats), CRHBP and RPL6 (both down-regulated in RLA-I vs. RHA-I animals) showed a similar tendency but a different order of magnitude of change among experiments; finally, NDN was validated neither in tendency nor in magnitude of change.     

Infantile stimulation and exploratory behavior in the rat

Rats were handled (H) or left non-handled (NH) prior to weaning. They were tested at approximately 35 days of age for 3 min/day, on 3 consecutive days, in a Y-maze having 1 black/white striped arm and 2 gray arms. Ambulation scores were significantly lower for NH animals which showed a greater preference for the striped arm as measured in terms of initial choice and time spent in that arm. The inference is that here NH animals exhibit a greater tendency to explore than H animals; the opposite conclusion would have pertained if ambulation had been used as the sole measure of exploration.     

Roman high- and low-avoidance rats: Present status of the swiss sublines,RHA/Verh and RLA/Verh,and effects of amphetamine on shuttle-box performance

Following a recent change in location and shuttle-box testing conditions, RHA/Verh and RLA/Verh rats were found to have maintained their characteristic patterns of avoidance (RHA/Verh) vs nonavoidance (RLA/Verh) behavior, regardless of the time of day tested. A second experiment demonstrated dose-related differences in the effects of amphetamine in both lines. In a third experiment it was seen that, whereas 2 mg/kg amphetamine was capable of increasing two-way avoidance acquisition in naive RLA/Verh rats to the levels of nondrugged RHA/Verh rats, this improvement was accompanied by substantial increases in spontaneous activity (intertrial responses). That learning was not genuinely enhanced by amphetamine was further indicated by the observation that the avoidance improvement disappeared when the drug was abruptly, or gradually, discontinued. The return toward control levels of avoidance and freezing responses was accompanied by a similar normalization of activity levels.     

Infantile handling and sex differences in shock-elicited aggressive responding of hooded rats

Infantile handled (Days 1-20) and nonhandled, male and female Long Evans hooded rats were tested at maturity (90-100 days) over 10 daily sessions for aggressive response to footshock. Individual jump and flinch thresholds for reactivity to shock, as well as paired aggressive responding to shock, were not significantly influenced by the handling procedure, although handled females ultimately adopted higher levels of fighting than nonhandled females. Handled rats were heavier than nonhandled rats of the same sex before and after testing for social aggression. Males fought significantly more than females; the discrepancy increased with additional sessions of paired exposure to shock. Male and female fight trends over sessions were linear and positive with a greater acceleration for males. The results were interpreted as indicative of a social learning variable occurring with repeated aggressive contact and primarily affecting males.     

Infantile stimulation and ultrasonic signaling: a possible mediator of early handling phenomena

Peromyscus maniculatus bairdi were either handled or not handled daily during the first 10 days of life. Handling consisted of removing the infant mice from the litter, placing them in small individual compartments where they remained for 3 min, and then replacing them in the nest. Immediately following the handling, simultaneous video and audio recordings were made of nest behavior for two 30-sec recording sessions. The nonhandled controls were treated comparably except that pups were never removed from the nest. Handled litters differed from nonhandled in terms of the average number of signals per recording session (132.5 vs. 16.5), the average signal duration (0.108 sec vs. 0.045 sec), and the average peak frequency of signals (22.216 kHz vs. 11.990 kHz). During those recording sessions in which no vocalizations were detected no physical interaction appeared between the mother and the infants. During all sessions in which vocalizations were emitted the mother was retrieving and grooming the infants. The data suggest a possible interpretation of the effects of early handling in terms of modified mothering, medidiated by modified vocalizations of infant mice as a function of the handling.     

Sympathetic hyperresponsiveness to hypothalamic stimulation in young hypertensive rats.

The possible occurrence of central sympathetic dysfunction during development of spontaneous hypertension was studied by recording aortic pressure and sympathetic nerve activity concurrently during electrical stimulation of the posterior hypothalamus in 9-wk-old Kyoto-Wistar rats. Even at this early age, basal levels for both measurements were already elevated significantly in those with spontaneous hypertension. Increases in sympathetic neural firing induced by graded hypothalamic stimulation were always followed by corresponding increases in blood pressure; magnitude of both effects was appreciably larger in spontaneously hypertensive than in normotensive rats, as was the vasodepression caused by blocking autonomic ganglia with pentolinium. By contrast, pressor responses to injected norepinephrine were almost equal thereby suggesting that cardiovascular reactivity was unaltered and that enhanced responsiveness to hypothalamic stimulation was directly due to the concomitant increase in sympathetic nerve activity. Although the exact site from which sympathetic hyperactivity originates was unidentified, our results support the interpretation that sympathetic mechanisms involving the posterior hypothalamus participate in elevating blood pressure during development of spontaneous hypertension in rats.     

Differential activation of dopamine release in the nucleus accumbens core and shell after acute or repeated amphetamine injections: a comparative study in the Roman high- and low-avoidance rat lines

The selectively bred Roman high- and low-avoidance rats differ in emotionality and responsiveness to the motor effects of acute and repeated psychostimulant administration. These lines also show drastic differences in the neurochemical responses of their mesolimbic dopamine systems to addictive drugs. The nucleus accumbens is critically involved in the locomotor activation produced by psychostimulants and in the augmentation of this effect observed upon repeated drug administration (i.e. behavioral sensitization), although there is not a general consensus as to whether the nucleus accumbens-core or the nucleus accumbens-shell is preferentially involved in such alterations. This study was designed to evaluate the effects of acute amphetamine (0.20mg/kg, s.c.) on dopamine output in the nucleus accumbens-shell and nucleus accumbens-core of the Roman lines under basal conditions (i.e. naïve rats) and after the repeated administration of amphetamine (1mg/kg, s.c.×10 days) or saline. We show that (1) in naïve rats, amphetamine caused a larger increment in dopamine output in the nucleus accumbens-shell vs the nucleus accumbens-core only in the Roman high-avoidance line; (2) repeated amphetamine elicits behavioral sensitization in Roman high-avoidance, but not Roman low-avoidance, rats; (3) in sensitized Roman high-avoidance rats, amphetamine provokes a larger increment in dopamine output in the nucleus accumbens-core, and an attenuated dopaminergic response in the nucleus accumbens-shell, as compared with Roman high-avoidance rats repeatedly treated with saline; and (4) such neurochemical changes are not observed in the mesoaccumbens dopaminergic system of the sensitization-resistant Roman low-avoidance line. We propose that (1) Roman high-avoidance and Roman low-avoidance rats differ in the vulnerability to develop psychostimulant sensitization, (2) the nucleus accumbens-core and nucleus accumbens-shell subserve distinct functional roles in this phenomenon, and (3) comparative studies in the Roman lines may provide insight into the influence of neural substrates and genetic background on the individual vulnerability to addiction.     

Characteristics of the pituitary-adrenal system in the syracuse high-and low-avoidance strains of rats (Rattus norvegicus)

After many generations of selective breeding, rats of the high-avoidance strain (SHA) average 67% avoidance responses in a two-way shuttle box, whereas those of the low-avoidance strain (SLA) average 0%. Adrenal gland weights, both absolutely and relative to body weight, are 40–50% greater in adult SLA than SHA rats of both sexes. Females of both strains have larger adrenal glands than males. Morphometry revealed that the difference in adrenal size in adults is entirely in the three cortical zones. The strain difference occurs as early as 21 days of age, whereas the sex difference appears only after puberty. A 2-min exposure to ether vapor induced an elevation in adrenal concentration of corticosterone which was significantly greater in SHA than SLA animals of both sexes at some time periods following the ether stress. Despite having smaller glands, older previously stressed SHA rats have higher basal adrenal concentrations of corticosterone than do SLA animals. The reduced steroidogenesis in the large adrenals of SLA rats suggests that there may be an enzymatic defect of genetic origin in those animals.This research was supported in part by David Ross Fellowships from the Purdue University Research Foundation to M. D. Isaacson and I. M. Rykaszewski and by Research Grant MH39230 to F. R. Brush from the National Institute of Mental Health.