Prolactin as a link between behavioral and immune differences between the Roman rat lines.

Roman high (RHA)- and low (RLA)-avoidance rats are two lines of Wistar rats genetically selected on the basis of their active avoidance behavior in a shuttle-box. They also differ in several other behavioral responses, such as their locomotor activity in novel environments (open-field, circular corridor), with the RHA rats being more active than the RLA animals, as well as in endocrine reactivity and immune functions. These experiments were designed to investigate further the neuroendocrine characteristics of these animals as a possible link between the brain and immune functions. Despite the marked behavioral and immune differences observed, no between-lines variation could be found in basal hypothalamo-pituitary-adrenocortical axis activity or in its responses to different protocols of novel environment stress, or after corticotropin-releasing factor (CRF) challenge. On the other hand, stimulated prolactin levels were higher in the low avoidance line. These results exclude the pituitary-adrenocortical axis and suggest prolactin as a link between behavioral and immune differences between the Roman lines. Moreover, these results indicate that these rats may be an excellent model for the study of the relationships between the brain and the immune system.     

Two-way avoidance behavior of the roman rat strains following brain lesions

Rats of the Roman strains, Roman High Avoidance (RHA), Roman Low Avoidance (RLA) and Roman Control Avoidance (RCA) were submitted to 5 conditioning sessions in a two-way shuttle box; the sessions were separated by a 24 hr-interval. Two indices were computed: a within-session progression index W and an inter-session progression index I. Index W was comparable in all three strains; index I was strongly negative in RLA and equal to zero or slightly positive in RHA rats. Lesion of the hippocampal commissure (HC) significantly facilitated CAR acquisition in RLA by making index I less negative: performances of RCA and RHA rats were unaffected. Lesions of the centro medianum parafascicularis nuclei of the thalamus (CM) significantly impaired acquisition of CAR in RCA and RHA by making index I more negative; performances of RLA rats were unaffected. Neither lesion altered index W. These results support the "freezing hypothesis" in the explanation of the Roman strain differences. They are in agreement with recent data on the emotional characteristics of the Roman strains and on the role of octopamine in avoidance behavior.     

Differential acquisition of a "working memory" task by the Roman strains of rats

Twenty-eight male rats of the Roman strains-fourteen RHA (Roman High Avoidance) and fourteen RLA (Roman Low Avoidance)-were submitted to a positively reinforced task, the delayed reinforced alternation test (DRA), in a T-maze. Performances of RLA rats were significantly better than those of RHA; RLA rats also had higher VTE (vicarious trial and error) and spontaneous alternation (SA) scores. These data confirm the fact that RLA may acquire positively reinforced learning as rapidly, or even more rapidly, than RHA rats, and that the differences in active avoidance behavior between these strains depend more on differential freezing behavior than on learning and memory capacities. Since the delayed reinforced alternation is considered as a working memory test, our results suggest that the Roman strains could be used as a genetic model for the neurobiological study of this form of memory.     

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)     

Effects of Cocaine and Morphine in Rats from Two Psychogenetically Selected Lines: A Behavioral and Brain Dialysis Study

The Swiss sublines of Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLA/Verh) rats differ in their reactivity to environmental and pharmacological stressors, in their sensitivity to stereotypies elicited by dopamine (DA)-mimetic agents, and in their densities of D₁ DA receptors in the terminal field of the mesoaccumbens DAergic projection, an important link in the neural networks involved in the motor effects and reinforcing properties of drugs abused by humans. The present study was therefore designed to compare the behavioral and neurochemical effects of cocaine (5 mg/kg, i.p.) and morphine (0.5 mg/kg, s.c.) in RHA/Verh and RLA/Verh rats. To this aim, we measured motor activity and DA output in the nucleus accumbens as determined by brain microdialysis. The number of counts corresponding to horizontal, vertical, and total motor activities accumulated in basal conditions during the 60-min acclimation period was significantly larger in RHA/Verh than in RLA/Verh rats. Moreover, horizontal, vertical, and total motor activities throughout the 120-min observation period that followed the administration of vehicle tended to be larger in RHA/Verh rats, although the difference between the two lines was not statistically significant. In RHA/Verh rats, locomotion, rearing, and total motor activity were significantly more intense after acute cocaine and morphine challenges than after vehicle administration, whereas no significant differences in motor activity were observed between control and cocaine- or morphine-treated RLA/Verh rats. No line-related differences were detected in the basal DA output, but the effect of cocaine on DA release was more robust in RHA/Verh rats. Likewise, the effect of morphine was more pronounced in RHA/Verh than in RLA/Verh rats. Because the mesoaccumbens DAergic pathway plays a central role in the acquisition of motivational valence by environmental and pharmacological stimuli and, therefore, in operant behavior, our results suggest that comparative behavioral and neurochemical studies in these two lines may provide useful information on the biological correlates of drug dependence.     

Psychophysiological profiles of the Roman strains of rats

Male rats of the Roman High and Roman Low Avoidance strains were submitted to four principal behavioral tests: food-motivated acquisition of bar-pressing in Skinner box, delayed reinforced alternation, locomotor activity in a "multibox apparatus," conditioned suppression, and two biological measures: blood pressure and brain octopamine level. Performances of RLA and RHA rats were significantly different in each behavioral test. Blood pressure was higher in RHA, but the difference only approached the 0.05 threshold. As previously reported, brain octopamine levels of RHA rats were significantly higher than those of RLA. A multivariate treatment (analysis of correspondences) was applied to the data of 10 behavioral tests. The main factor extracted by this treatment clearly separated the two strains. Among the variables which best differentiate RHA and RLA rats, several do not involve stress (working memory, acquisition of bar-pressing, and locomotor activity).     

Infantile (handling) stimulation and behavior in young Roman high- and low-avoidance rats.

The effect of infantile handling stimulation on exploratory and emotional behavior of Roman high- and low-avoidance (RHA/Verh and RLA/Verh) weanling rats was investigated. Postnatally handled and nonhandled, 4-week-old males and females from both psychogenetically selected lines were exposed to a hexagonal tunnel maze, including an illuminated central arena. Postnatal handling increased exploratory behavior and decreased emotional reactivity as expressed by increased entries into the central arena and a reduction in defecations in both lines of rats. These effects were more pronounced in the RLA/Verh rats. In agreement with earlier studies using nonselected adult rats, the females of both lines (especially those from the RHA/Verh line) were more sensitive than males to the positive influences of early stimulation.     

Biochemical Parameters of Dopaminergic and GABAergic Neurotransmission in the CNS of Roman High-Avoidance and Roman Low-Avoidance Rats

The dopaminergic (DAergic) and GABAergic pathways in the central nervous system (CNS) are involved in the control of emotions, in the reactivity to stressful stimuli, and in the positive and negative reinforcing properties of psychotropic drugs. In the present review, we summarize the differences in a range of neurochemical markers of GABA- and DA-mediated neurotransmission in the CNS of Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLA/Verh) rats, two psychogenetically selected lines that differ in what may be considered to be level of emotionality. The stimulatory effect of GABA on ³⁶Cl^– uptake was less pronounced in the cerebral cortex of RLA/Verh rats compared to RHA/Verh rats. In addition, the binding affinity of [³⁵S]TBPS, a selective ligand of the convulsant site located in the chloride channel of GABA_A receptors, was significantly lower in the hippocampus of RLA/Verh rats than in their high-avoidance counterparts. On the other hand, the density of D₁ DA receptors labeled with [³H]SCH 23390 was lower in the nucleus accumbens of RLA/Verh rats compared to RHA/Verh rats. Brain microdialysis studies demonstrated that tail-pinch stress and subconvulsant doses of the anxiogenic compound pentylenetetrazol increased the extracellular concentrations of DA in the prefrontal cortex of hypoemotive RHA/Verh rats but not in their hyperemotive RLA/Verh counterparts. These line-dependent differences in GABAergic and DAergic neurotransmission may contribute to the distinct emotionality and responsiveness to centrally active drugs of RHA/Verh and RLA/Verh rats.     

Neuroendocrine Correlates of Emotional Reactivity and Coping in Male Rats from the Roman High (RHA/Verh)-and Low (RLA/Verh)-Avoidance Lines

The Roman high (RHA/Verh)-and low (RLA/Verh)-avoidance rats, originally selected and bred for rapid vs. poor acquisition of a two-way active avoidance response, differ in emotional reactivity and sensitivity to stressors in various other test situations. These behavioral differences are associated with particular neuroendocrine and neurochemical characteristics. The aim of this short review is to present data currently available on the neuroendocrine profiles of RHA/Verh and RLA/Verh rats, together with more recent findings which suggest that differences in peripheral and central hormonal responses, and in hormone action on the brain, may be closely related to emotional reactivity and coping ability. Although genetic factors certainly play a major role, there is also evidence that epigenetic factors, e.g., early environmental influences, can modulate the phenotypic expression of the basic behavioral and neuroendocrine traits characterizing these lines. These psychogenetically selected lines can therefore be used as a model to investigate interactions between genes and the environment in determining each individual's sensitivity to stress and coping abilities (vulnerability model). This model may prove particularly useful for studies on the etiology and pathophysiology of anxiety and affective disorders and their neuroendocrine correlates.     

Diverse Sensitivity of RHA/Verh and RLA/Verh Rats to Emotional and Spatial Aspects of a Novel Environment as a Result of a Distinct Pattern of Neuronal Activation in the Fear/Anxiety Circuit

Psychogenetically selected Roman high (RHA/Verh) and Roman low (RLA/Verh) avoidance rats constitute a well-recognized model of diverse emotional reactivity. The two Swiss lines display marked behavioral and endocrine differences in reaction to a novel environment. In our study we found that these differences are accompanied by a distinct, line-specific pattern of neuronal activation within the fear/anxiety circuit. We have compared the c-Fos protein expression in the medial prefrontal cortex (mPFC), basolateral (BLA), central (CeA), medial (MeA), and cortical (CoA) nuclei of amygdala, paraventricular nucleus of the hypothalamus (PVN), and CA1, CA2, and CA3 fields of the hippocampus upon exposure to a novel situation of different stressorgeneity (open field with illuminated center, elevated plus maze, hole board test and acute restraint). Profound between-line differences in the sensitivity to emotional and spatial aspects of the behavioral challenge were observed for tests measuring spontaneous behavior. This effect seems to reflect different motivational factors driving the rat behavior, which clearly suggests that the diverse emotional reactivity of RHA/Verh and RLA/Verh rats is a result of different activation of the fear/anxiety circuit. Edited by Stephen Maxson.     

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.     

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.     

Correlated Behavioral Traits in Rats of the Roman Selection Lines

The current theories of animal personality are based on the observation that individual variation in behavior and physiology appears to be consistent across contexts. Rats of the Roman selection lines have been originally selected for differences in shuttle-box behavior. Besides differences in active avoidance, these animals differ more generally in coping style. Roman high avoidance (RHA) rats show high levels of active avoidance, whereas Roman low avoidance (RLA) rats tend to respond with a more passive (i.e. freezing) response. Based on the two tier model of coping styles, we hypothesized that RHA rats would show high levels of offensive behavior and are more impulsive compared to RLA rats. We characterized animals in a two-way active avoidance task on five consecutive days. Thereafter animals were tested for their level of offensive aggression and impulsive behavior. The level of offensive aggression was examined in a standard resident-intruder paradigm. Furthermore, we tested aspects of impulsivity in an unpredictable operant conditioning paradigm (variable interval-15 schedule) for food reinforcement and during extinction of lever press behavior. We show that RHA rats are indeed characterized by high levels of two-way active avoidance in a shuttle-box paradigm. Surprisingly, the level of offensive aggression was higher in RLA compared to RHA rats. Consistent with the coping style interpretation, the number of lever presses in the VI-15 schedule for food reinforcement was higher in RHA rats compared to RLA rats. During a session of frustrating non-reward, RHA rats were more persistent. Taken together, results of the two-way active avoidance task and VI-15 performance in rats of the Roman selection lines fit with the two tier model of coping styles. Unexpectedly, the level of offensive aggression does not match with this model.     

Contribution of the Roman strains of rats to the elaboration of animal models of memory.

Performances of male rats of the Roman High (RHA)- and Roman Low (RLA)-Avoidance strains were compared along four essential dimensions: working memory, reference memory, spontaneous locomotion and avoidance conditioning. Performances of RLA and RHA rats were significantly different for each dimension. As constantly reported, RHA rats were by far superior to RLA in avoidance conditioning. They had also higher levels of locomotor activity. On the opposite, RLA performed better than RHA in an appetitive working memory task, the delayed reinforced alternation, and were also superior in an appetitive reference memory task, the 5-unit linear maze. These results confirm the fact that RLA rats may acquire positively reinforced learning more rapidly than RHA rats and that the differences in active avoidance behavior between the two strains depend more on differential freezing behavior than on learning or memory capacities. Beyond the problem of the characterization of the Roman strains, these data might give indications on the relationships between behavioral tests widely used in rats, and on their use as memory models.     

Corticotropin-Releasing Hormone Modulation of a Conditioned Stress Response in the Central Amygdala of Roman High (RHA/Verh)-Avoidance and Low (RLA/Verh)-Avoidance Rats

Roman high (RHA/Verh)- and low (RLA/Verh)-avoidance rats are selected and bred for rapid versus nonacquisition of two-way, active avoidance behavior in the shuttle box. RHA/Verh rats generally show a more active coping style than do their RLA/Verh counterparts when exposed to various environmental challenges. The central nucleus of the amygdala (CeA) is known to be involved in the regulation of autonomic, neuroendocrine, and behavioral responses to stress. Its involvement in the selection of coping strategies has also been suggested. Corticotropin-releasing hormone (CRH) seems to be one of the key neurohormones in the control of CeA output. Neuroanatomical studies have revealed that the majority of CRH fibers from the CeA have direct connections with autonomic regulatory nuclei in the brain-stem, e.g. lateral parabrachial nucleus (lPB). The effects of CRH (30 ng) on modulating CeA activity were studied by infusion of CRH into the CeA during conditioned stress (inescapable foot-shocks) in RHA/Verh and RLA/Verh male rats. Heart-rate responses after CRH treatment were not changed in either line. However, distinctly different behavioral responses were seen after CRH infusion into the CeA of both rat lines. A decrease in immobility responses was seen in both RHA/Verh and RLA/Verh rats, while an increase in exploration was observed in RHA/Verh rats only in the conditioned stress situation. Rearing levels were increased in the RHA/Verh rats, whereas they were decreased in the RLA/Verh animals. As a result of CRH infusion, the number of FOS immunoreactive cells in the lPB of RLA/Verh rats was decreased, whereas an opposite response was found in RHA/Verh rats. These results indicate that the CRH system of the CeA connected with output brain-stem areas is differentially involved in the cardiovascular and behavioral responses of these rats having different coping styles.     

A Comparative Study of the Behavioral Patterns of RLA/Verh and RHA/Verh Rats in the Exploration Box

Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats differ in their emotionality profiles. As emotional processes are considered to play an important role in exploratory behavior, differences between the Roman sublines in responding to a novel environment can be anticipated. The present study provides a quantitative (based on the frequency of particular responses) and qualitative (based on the frequency of behavioral sequences) comparison of exploratory behavior of RHA/Verh and RLA/Verh rats. Rats of both sexes were tested individually for 30 min in an exploration box containing a few objects. Observation periods were divided, based on earlier studies, into three consecutive blocks of 5, 10, and 15 min, respectively. Analysis of quantitative data showed differences in sniffing (RHA/Verh scoring higher) and in immobility (RLA/Verh scoring higher), both persisting throughout all measurement periods. Males of both sublines were generally more active and exploratory than their respective females. The analysis of sequential data revealed that during the first 5 min RHA/Verh rats showed a less diverse but more exploratory repertoire and that during the first 15 min males of both sublines showed more behavioral sequences than females. This study indicates that the Roman rat sublines should provide a useful model to study the mechanisms of exploratory behavior.     

Selection for two-way avoidance deficit inhibits shock-induced fighting in the rat

Matched pairs of male and female, Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats were tested for shock-induced fighting, and single rats were tested for comparative sensitivity to footshocks. The main finding was that the RLA/Verh rats showed a total absence of shock-induced fighting or posturing. The females of both selected lines showed lower flinch-, shuffle- and jump-shock thresholds than did the males. Although the RHA/Verh rats had lower shuffle and jump thresholds than did the RLA/Verh rats, there was no significant difference in the initial detection of foot-shocks by either group (flinch threshold), and all 3 levels of threshold values for both were well below the 3 mA shock level used in the shock-induced fighting experiment. It was concluded that the genetically-based response suppression (freezing behavior) seen with RLA/Verh rats under these and all shock-stress conditions played a greater role in the inhibition of shock-induced fighting than did sensitivity to footshocks per se.     

Impulsivity and aggressive behavior in Roman high and low avoidance rats: baseline differences and adolescent social stress induced changes

Adverse and stressful experiences during adolescence are often of a social nature. The social defeat model in rats is used as an animal model for bullying in humans. Usually large individual differences in response to social defeat are found. The personality type that is mostly affected and the underlying mechanisms are unknown. We used male rats of the Roman selection lines to test whether social defeat (between postnatal days 45 and 57) followed by social isolation has a different impact in animals with divergent levels of emotional reactivity and coping style. The level of offensive aggression, impulsivity and performance during frustrating non-reward (extinction) were used as measures for the adult coping style of animals. Impulsivity was measured by performance on an unpredictable operant conditioning schedule (variable interval-15, VI-15) for food reinforcement. This study demonstrates that the adult, baseline level of impulsivity is higher in Roman high avoidance (RHA) rats. RHA rats showed a higher number of lever presses compared to Roman low avoidance (RLA) rats on a VI-15 schedule. The level of offensive aggression did not differ between the two lines. Surprisingly, a tendency towards more offensive aggression in RLA rats was found. Social stress during adolescence disturbed the normal development of adult personality, mostly in RHA rats. RHA rats that were defeated during adolescence reduced the number of lever presses on the VI-15 schedule of reinforcement and were more persistent during a session of frustrating non-reward. However, we did not find an effect of social defeat on performance during extinction. A tendency towards increased attack latencies after social defeat in adolescence was found. The time spent on offensive aggression was unaffected by social defeat.     

Differential Circadian Eating Patterns in Two Psychogenetically Selected Strains of Rats Fed Low-, Medium-, and High-Fat Diets

Spontaneous eating patterns in male, inbred Roman high- and low-avoidance rats (RHA/Verh, RLA/Verh) were continuously recorded while animals were successively offered three isocaloric (16.5-kJ/g) diets: a low-fat, high-carbohydrate diet (LF; 3.3% fat), a medium-fat diet (MF; 18% fat), and a high-fat diet (HF; 40% fat), the latter being followed once again by the LF diet. Under the conditions of this experiment, overall 24h food intake did not differ significantly between RHA/Verh and RLA/Verh rats, but was significantly higher for both rat strains on the MF and HF diets than on the LF diet. Despite the similar 24h-food intake, RHA/Verh rats ate transiently less than RLA/Verh rats during the third quarter of the dark phase under all dietary conditions. These differences were due to the RHA/Verh rats' longer intermeal intervals (with all diets) and smaller meals (with the MF and HF diets) and were compensated for during the last 3 h of the dark phase. On the LF diet, dark-phase meal frequency was higher and both nocturnal meal size and mean eating rate within meals were lower in RLA/Verh rats than in RHA/Verh rats. With the MF and HF diets, mean nocturnal meal size and meal duration were higher and mean eating rate was lower in RLA/Verh rats than in RHA/Verh rats. For both strains, nocturnal meal size was significantly higher with the MF and HF diets than with the LF diet, and nocturnal meal frequency was lower with the HF diet than with the other two diets. Although body weights were similar at the start of the study, RLA/Verh rats gained significantly more weight than did RHA/Verh rats by the end. As has often been the case with other aspects of behavior studied, differences in neuromodulatory systems (e.g., serotoninergic and dopaminergic) between RHA/Verh and RLA/Verh rats may directly or indirectly contribute to the subtle differences in eating patterns observed here.     

Neonatal Handling and Environmental Enrichment Effects on Emotionality, Novelty/Reward Seeking, and Age-Related Cognitive and Hippocampal Impairments: Focus on the Roman Rat Lines

Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats are selected and bred for extreme divergence in two-way active avoidance acquisition. In addition, compared to RLA/Verh rats, RHA/Verh rats are (behaviorally and physiologically) less anxious or reactive to stressors, show increased novelty (sensation)-seeking behavior as well as a higher preference for rewarding substances, and are usually less efficient in learning tasks not involving shock administration. The present article reviews evidence showing that neonatal handling and/or environmental enrichment leads to enduring effects (their magnitude frequently depending upon the rat line) on those behaviors. For example, it has been found that neonatal handling reduces most of the (behavioral and physiological) signs of emotionality/anxiety in RLA/Verh rats, while environmental enrichment increases their novelty seeking (also the case with RHA/Verh rats), saccharin and ethanol intake, and sensitivity to amphetamine. Finally, initial results (currently being further elaborated upon) support a preventive action of both environmental treatments on age-related impairments in learning a spatial, water maze task as well as on hippocampal neuronal atrophy.