Roman Low Avoidance Research Articles

Environmental enrichment and postnatal handling prevent spatial learning deficits in aged hypoemotional (Roman high-avoidance) and hyperemotional (Roman low-avoidance) rats.

The present study investigated the enduring effects of postnatal handling (administered during the first 21 days of life), and environmental enrichment (for a period of 6 months starting 3 weeks after weaning) on spatial learning in 24-month-old hypoemotional (Roman high-avoidance, RHA/Verh) and hyperemotional (Roman low-avoidance, RLA/Verh) rats. Two groups of 5-month-old rats from both lines were also included in the experiment as young controls. The Roman lines performed differently in the Morris water maze: Path lengths of RLA/Verh rats were shorter and they swam at lower speed than RHA/Verh rats, showing quicker and more efficient learning overall. Postnatal handling improved learning mainly in RHA/Verh rats, whereas environmental enrichment was able to prevent the deficits shown by aged controls of both lines. Young, enriched, and handled plus enriched animals exhibited better performance than impaired aged controls, to the point that aged enriched and handled plus enriched animals did not differ from young controls. Thus, besides indicating that RLA/Verh rats are better learners than RHA/Verh rats in the Morris water maze, this study demonstrates that environmental enrichment prevents the cognitive loss associated with aging, over the long term. Finally, the positive effects obtained with postnatal handling were dependent on the rat line.

Effects of training, early handling, and perinatal flumazenil on shuttle box acquisition in Roman low-avoidance rats: Toward overcoming a genetic deficit

The present series of studies investigated the effects of intensive training, postnatal handling-stimulation and/or perinatal flumazenil (Ro 15-1788, benzodiazepine receptor antagonist) on the acquisition of two-way active avoidance by Roman low-avoidance (RLA/Verh) rats. This rat line has been selectively bred for poor avoidance in the shuttle box, while their Roman high-avoidance counterparts (RHA/Verh) have been selectively bred for their extremely good performance in that task. In the first experiment, RLA/Verh rats submitted to a long and intensive training procedure (unlike those submitted to short training) were able to achieve a performance of 56% of avoidances per session. In the second experiment both postnatal handling and perinatal flumazenil treaments increased avoidance responding in another group of RLA/Verh rats tested at the age of 18 months. Finally, in the last experiment, the performance of a third stock of RLA/Verh rats of the same age which had received perinatal flumazenil did not differ, on the later phases of training, from that shown by RHA/Verh animals. The results are discussed in terms of the “warm up” phenomena which seems to be highly involved in the selection of RLA/Verh rats, as well as on the possibility that central benzodiazepine receptors could play a role in the genetic deficit shown by RLA/Verh rats, which apparently confers a greater emotivity.

Cocaine sensitivity in roman high and low avoidance rats is modulated by sex and gonadal hormone status

Repeated exposure to stressful stimuli or psychostimulants increases sensitivity to the motoric effects of these drugs, i.e. behavioral sensitization. The objective of the present experiment was to determine the interaction between factors that modulate psychostimulant sensitivity, i.e. sex and circulating gonadal hormone levels, in rats with a genetically distinct locomotor response to novelty: Roman Low Avoidance rats (RLA) freeze while Roman High Avoidance rats (RHA) remain motorically active. Ninety-six male and female RHA and RLA rats were gonadectomized (GDX) just after weaning or as adults, or left gonadally intact. Each rat received a total of 9 injections of cocaine hydrocholoride (10 mg/kg, IP), at 3–4-day intervals for 5 weeks. Locomotor activity was measured after each injection, and stereotypies were rated 1×/week. Open field behavior (10 min) and plasma corticosterone were measured 2 weeks after the final injection. Overall, the RHA line was more sensitive to (1) cocaine's stereotypic effects, and (2) the influence of ovarian hormones on the cocaine's acute and sensitizing effects on locomotor activity. Therefore, genetic background not only determines cocaine sensitivity, but also the influence of gonadal hormones on locomotor activity. These interactions are relevant when considering the genetic contribution to abuse liability.

Maturation of the behavioral and neuroendocrine differences between the Roman rat lines

The behavioral and/or neuroendocrine reactivity to psychological (open-field exposure) and physiological (CRF challenge) stimulations, as well as adrenal tyrosine hydroxylase (TH) and phenylethanolamine N-methyl transferase (PNMT) activities were measured, at different ages, in the Roman high avoidance (RHA) and Roman low avoidance (RLA) rat lines that have been genetically selected on the basis of their divergent active avoidance behavior. The highest locomotor activity in the open field, associated to blunted prolactin and renin reactivity to an emotional stress and lower specific TH and PNMT activities, characterized the RHA rats of all ages. HPA axis reactivity to psychological and/or physiological stimulations was identical in young animals (14 weeks old) of the two lines. Nevertheless, it displayed with age maturation processes, since the amplitude of postopen-field secretion peak for ACTH was larger in RLA rats from 20 weeks on, the response to CRF being not increased until 42 weeks. These maturation processes could result from genetically influenced changes related to environmental stimulations. Therefore, the Roman lines may be an excellent model to study the interactions between the genetic and developmental factors controlling the coupling between both behavioral and neuroendocrine functions.

GABAergic and dopaminergic transmission in the brain of Roman high-avoidance and Roman low-avoidance rats

The GABAergic and dopaminergic pathways in the central nervous system (CNS) play a pivotal role in the control of emotions and in the adaptive responses to stressful stimuli. The present study was aimed at characterizing a range of biochemical markers of GABA- and dopamine-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 their level of emotionality. The stimulatory effect of GABA on 36Cl − uptake was less pronounced in the cerebral cortex of RLA/Verh rats as compared to RHA/Verh rats, whereas no line-related changes were detected in [ 3H]GABA and [ 3H]flunitrazepam binding. On the other hand, the density of D 1 dopamine receptors labeled with [ 3H]SCH 23390 was lower in the nucleus accumbens of RLA/Verh rats as compared to their RHA/Verh counterparts, whilst no line-dependent changes were observed in the binding parameters of D 1 dopamine receptors in the striatum, amygdala, and prefrontal cortex. These biochemical differences may contribute to the distinct emotionality and responsiveness to the effects of psychoactive drugs of RHA/Verh and RLA/Verh rats.

Augmenting and reducing of visual evoked potentials in Roman high- and low-avoidance rats

Human and cat high sensation seekers tend to show increasing amplitudes (augmenting) of the P 1 and N 1 components of the visual evoked potential (VEP) to increasing intensities of light flash, whereas low sensation seekers show VEP reducing. Roman high-avoidance (RHA) and Roman low-avoidance (RLA) rats have behavioral traits comparable to human and cat high and low sensation seekers, respectively. RHA rats show greater exploration, activity, and aggression than do RLA rats. Rats of each Roman line and Wistar rats were anesthetized with chloral hydrate and maintained at a stable moderate anesthetic level. VEPs to each of five flash intensities were computer averaged for each rat. The slopes of P 1 amplitudes as a function of flash intensity were significantly greater in the RHA than the RLA rats. RHA rats were clear augmenters; RLA rats had almost flat amplitude-intensity functions. The Wistar rats had slope functions that were similar to those of the RLA rats. This study shows that the relationship between sensation-seeking behavior and VEP augmenting and reducing has a heritable base and extends across species from human, cat, and rat. In addition, we demonstrate a rat model of this relationship that yields advantages of genetic homogeneity and a short generational time, and provides access to a wealth of behavioral data and experimental manipulations available for the rat.

Posttraining norepinephrine infusion into the central amygdala differentially enhances later retention in Roman high-avoidance and low-avoidance rats.

Memory-enhancement effects of norepinephrine (NE) were investigated by infusing NE into the central amygdala (CEA) of Roman high-avoidance (RHA) and low-avoidance (RLA) rats after training on active and passive behaviors in the defensive-burying paradigm. During acquisition, both lines spent comparable time in burying behavior. RLA rats, but not RHA rats, also displayed substantial immobility. During retention, the cerebrospinal-fluid-treated RLA rats mostly displayed immobility to the nonelectrified probe, whereas the RHA rats showed neither burying nor immobility. In the RLA rats, high-dose (200 ng), but not low-dose (20 ng), NE infusion enhanced the duration of the passive response (immobility) without affecting the active response. NE given into the CEA of RHA rats caused a selective dose-dependent appearance of the active behavioral component. The results suggest a phenotype-dependent effect of intra-amygdaloid NE on memory processes in the rat.

Opposite effects of central amygdaloid vasopressin and oxytocin on the regulation of conditioned stress responses in male rats.

The central nucleus of the amygdala (CEA) seems to be selectively involved in the passive component of the behavioral (immobility) and the accompanying parasympathetic response (bradycardia) during conditioned environmental challenges,' leaving conditioned sympathetic, adreno-medullary, and adreno-cortical responses unaffected (Roozendaal et al., in preparation). Robust monosynaptic, .peptidergic connections to the vagal complex and periaqueductal gray may be mediating these effects. Intracerebroventricularly administered arginine-8-vasopressin (AVP) potentiates these stress-induced bradycardiac and immobility responses.* AVP may exert these effects via AVP and/or oxytocin (OT) receptive systems in the CEA.' Microinfusion of AVP (dissolved in 1 p1 artificial CSF) into the CEA of conscious male Wistar rats under resting conditions, leads to a dose-dependent change in heart rate and behavior: A high dose of AVP (2 ng), but not lower ones (20 and 200 pg), causes a transient increase in heart rate and behavioral a~t iv i ty .~ Moreover, in a part of the animals, the low dose of AVP even causes an opposite effect, that is, bradycardia. OT administration (2 ng) induces responses similar to those induced with the high dose of AVP. Neither plasma norepinephrine nor epinephrine is influenced after AVP infusion, suggesting that the tachycardia is due to reduced parasympathetic control, rather than a stimulation of the sympathetic outflow. The effects of AVP and OT during conditioned stress were studied in male Roman high-avoidance (RHA) and Roman low-avoidance. (RLA) rats, selected genetically for shuttle-box acquisition behavior. In RLA rats the cardiac and behavioral responses to the emotional stressor are bradycardia and immobility,

The Roman strains of rats as a psychogenetic tool for pharmacological investigation of working memory: example with RU 41656.

This study examined the effects of RU 41656, a dopaminergic D2 agonist, on the differential working memory performances and on the differential activities of the neurochemical systems of the Roman high (RHA) and Roman low (RLA) avoidance strains of rats. Compared with RLA, RHA performed worse in three tests of working memory (spontaneous alternation, radial maze and object recognition) and had higher levels of exploratory locomotor activity. Hippocampal and frontal cortex choline acetyltransferase (ChAT) activities were lower in RHA. Frontal cortex DA and DOPAC levels, hippocampal and striatal 5-HT and NA levels were higher in RHA. RU 41656 induced a significant improvement in working memory performance of RHA, whereas in RLA it had no effect. It decreased exploratory locomotor activity in both strains. ChAT activity in hippocampus was not affected by RU 41656 in either strain, whereas in frontal cortex it was increased in RHA but not in RLA. Hippocampal NA levels were decreased by RU 41656 in RHA but not in RLA. These results confirm previous data concerning the promnesic effect of RU 41656 and extend the finding that the Roman strains are a psychogenetic model for the behavioural, neurochemical and psychopharmacological study of the working memory in rats.

Low-level cadmium exposure increases one-way avoidance in juvenile rats

Twenty-seven pregnant rats from three genetic lines, Roman High Avoidance (RHA), Roman Low Avoidance (RLA), and Satinder's Heterogeneous Stock (SHS), received daily SC injections of either 0.075 mg/kg CdCl 2 (low dose), 0.225 mg/kg CdCl 2 (high dose), or an equivalent volume of saline vehicle (control) throughout gestation. Cd-exposed progeny from the RHA genetic line weighed significantly less than RHA control progeny (pd 35–44); however, SHS progeny from the low-dose group weighed significantly more than progeny from any other group (pd 14–44). Unconditioned escape response (UER) level was determined on pd 39. Progeny from the high-dose group required a significantly lower UER level as compared to the low-dose group. Acquisition of conditioned avoidance responses was tested from pd 41 to 44. There were significant differences due to dose for one-way avoidance responses. Cd-related differences in one-way avoidance were restricted to progeny from the SHS genetic line. SHS progeny from the high-dose group demonstrated significantly more one-way responses when compared to the control group. Differences in avoidance responses are discussed in relation to Cd-induced hyper-nociception. The concentration of Cd in the liver and kidney of dams and progeny was determined by Inductively Coupled Plasma spectroscopy (ICP). There were significant dose-related differences in Cd concentration in maternal tissues.

Strain differences in rats with respect to speed of conflict resolution

Speed of conflict resolution was studied in a conditioned punishment paradigm in a Skinner box and a straight runway. In both experimental situations speed of conflict resolution was defined as the latency to gain food during an approach-avoidance conflict. In the Skinner box Tryon Maze Bright rats were faster in speed of conflict resolution than Tryon Maze Dull rats, and Roman Low Avoidance rats were faster than Roman High Avoidance rats. In the runway situation, Wistar Kyoto rats were faster in solving the conflict than randomly bred Wistar Wu rats and Brown Norway rats were faster than Wistar Wu rats. Differences between the strains in speed of conflict resolution could not be consistently explained from strain differences in approach or avoidance behavior, measured separately. It is, therefore, suggested that speed of conflict resolution is a unique parameter.

The pineal complex in Roman high avoidance and Roman low avoidance rats

Previous studies have shown that the pineal gland of Roman high avoidance (RHA/Verh) rats is larger than that of Roman low avoidance rats (RLA/Verh). In the present study measurement of enzyme activities (serotonin-N-acetyl-transferase, hydroxyindole-O-methyltransferase) revealed that pineals of RHA/Verh rats are twice as active in melatonin production than pineals of RLA/Verh rats. Indoleamine content was also higher in RHA/Verh rats, whereas noradrenaline content was the same in both lines. When values were expressed per mg protein, these differences disappeared except for N-acetyl-serotonin and noradrenaline which were higher or lower in RHA/Verh rats, respectively. Both lines had higher serum levels of melatonin during the dark phase than during the light phase. However, RHA/Verh rats had increased serum levels as compared to RLA/Verh rats during both day and night. Morphometric analysis of the deep and superficial part of the pineal complex revealed, that the volumes of both parts are enlarged in RHA/Verh rats. Electron microscopic studies of pineals collected during day- and nighttime showed higher numbers of synaptic ribbons per unit area in pineals of RHA/Verh rats. In pineals collected during June synaptic ribbons displayed a day/night rhythm in RHA/Verh rats only, whereas in glands of both lines collected during November no daily changes were found. These results show that closely related but divergently selected rat lines may differ in pineal ultrastructure and pineal function.

Prenatal Ontogenesis of Brain Phenolamines and Catecholamines in Relation to Their Metabolizing Enzymes in Roman Avoider Strains of Rats

Phenolamines, particularly octopamines, are of special importance in avoidance behavior. In the Roman low avoidance (RLA) strain, p-octopamine can induce locomotor behavioral activity that is normally observed in the Roman high avoidance (RHA) strain. For these reasons, the levels of prenatal octopamines (para and meta isomers) have been studied in relation to noradrenaline and dopamine levels. In the hypothalamus and brainstem of RHA, a maximum level of the para isomer is observed at 15 days of embryonic development but, unlike in controls and RLA animals, this level remains almost constant until 20 days. For the meta-isomer and catecholamines, there is a 1-2 day delay in detection between controls and RLA or RHA. The study of related enzyme activities reveals that tyrosine hydroxylase displays a 2-day delay in RHA when compared to the control value at 19 days of fetal life. These results are discussed in terms of the role of p-octopamine in avoidance conditioning and of the possible delayed expression of the tyrosine hydroxylase gene in Roman strains of rats.

Genetic and environmental influences on behavioral and neurochemical aspects of emotionality in rats

Three pairings of rats (two derived from divergent, selective breeding and one from divergent environmental conditions) were compared with regard to behavioral and hormonal parameters. Striking differences were observed: results obtained in our own laboratory as well as those found in a review of the literature pointed to higher emotionality (e.g., increased defecation and corticosterone secretion, etc.) in Roman low-avoidance, Wistar-Kyoto and group-housed rats, as compared to their respective counterparts, Roman high-avoidance, spontaneously hypertensive, and individually housed Wistar rats. Concomitant receptor binding studies reviewed here (3H-diazepam- and 3H-imipramine-binding sites) have revealed, however, less consistent intrapair differences.

Stressful environmental stimuli increase extracellular DOPAC levels in the prefrontal cortex of hypoemotional (Roman high-avoidance) but not hyperemotional (Roman low-avoidance) rats. An in vivo voltammetric study

The effects of a variety of stressful environmental situations on dopamine metabolism in the prefrontal cortex (as assessed by in vivo voltammetry with carbon fiber electrodes) have been compared in two genetically selected lines of rat (Roman high (RHA/Verh) and low (RLA/Verh) avoidance) which differ drastically in their level of emotionality. Heart rate was continuously monitored in these animals (via chronically implanted subcutaneous electrodes) so as to index the emotional reaction to the stressors. An electrochemical signal corresponding to the oxidation of dihydroxyphenylacetic acid (DOPAC) was recorded in the deeper laminae of the anteromedial prefrontal cortex in both lines of rats. Under normal condition, this signal was stable for at least 4 h and its amplitude was similar in both lines. Introduction of the animals into an unfamiliar environment (30 min), application of a mild tail pinch (10 min) or of a high-intensity loud noise (30 min) or immobilization (20 min) were all associated with an increase in extracellular cortical DOPAC levels in the hypoemotional RHA/Verh line but not in the hyperemotional RLA/Verh line. Similarly, forced locomotion on a rotarod (40 min) provoked a dramatic increase in the amplitude of the cortical DOPAC oxidation peak in RHA/Verh rats and only a mild increase in this parameter in RLA/Verh rats. In RHA/Verh rats, tolerance to this increase was observed when animals were subjected to forced locomotion every day for 5 days. All of the stressful situations investigated provoked an immediate augmentation of heart rate which resumed gradually after cessation of the stressful stimulus; the magnitude and duration of this increase were much greater in RLA/Verh than in RHA/Verh rats. Moreover, in all stress situations, RLA/Verh but not RHA/Verh rats showed behavioral signs of emotional response e.g. defecation, freezing and self-grooming. It is concluded that the increase in cortical dopamine metabolism induced by stress is not connected to the emotional reaction caused by the aversive nature of the stressor but may rather reflect a heightened attention of the animal or activation of cognitive processes in an attempt to cope with the stressor.

Oxytocin stress responses are dependent upon emotionality

Comparison of posterior pituitary responses to stress in rat strains with contrasting emotionality has revealed an inverse relationship between oxytocin (OT) responses and emotional reactivity. The plasma OT and arginine-vasopressin (AVP) responses to stress were determined in two psychogenetically selected strains of rat, Roman high avoidance (RHA) and Roman low avoidance (RLA), RLA's being more emotionally reactive than RHA's. Following 1 min immobilisation stress, plasma levels of OT were significantly higher in RHA's compared to RLA's. This finding correlates with the previous demonstration of a sex difference in the OT stress response, females having a higher response than males. Plasma levels of AVP were not significantly modified by immobilization in either strain of rat. However, control levels of AVP were markedly raised in both male and female RHA's.

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).

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.

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.

Genotypic differentiation of Roman selection lines did not "disappear" with inbreeding.

A recent claim that inbred sublines of the Roman high and low selection strains are not differentiated for avoidance performance casts doubt on the results of an earlier triple test-cross study using the lines. However, data from 220 litters of crosses to these lines demonstrate that the claim was certainly incorrect at the time of the earlier study, 1975–1977. The anomalous results which gave rise to the claim do not fit well with our knowledge of the genetic architecture of avoidance conditioning in rats; it is unlikely that inbreeding per se results in rapid acquisition of twoway active avoidance in Roman Low Avoidance rats.