Acquisition Of Two-way Avoidance Research Articles

Conservation of Phenotypes in the Roman High- and Low-Avoidance Rat Strains After Embryo Transfer

The Roman high- (RHA-I) and low-avoidance (RLA-I) rat strains are bi-directionally bred for their good versus non-acquisition of two-way active avoidance, respectively. They have recently been re-derived through embryo transfer (ET) to Sprague-Dawley females to generate specific pathogen free (SPF) RHA-I/RLA-I rats. Offspring were phenotyped at generations 1 (G1, born from Sprague-Dawley females), 3 and 5 (G3 and G5, born from RHA-I and RLA-I from G2-G4, respectively), and compared with generation 60 from our non-SPF colony. Phenotyping included two-way avoidance acquisition, context-conditioned fear, open-field behaviour, novelty-seeking, baseline startle, pre-pulse inhibition (PPI) and stress-induced increase in plasma corticosterone concentration. Post-ET between-strain differences in avoidance acquisition, context-conditioned freezing and novelty-induced self-grooming are conserved. Other behavioural traits (i.e. hole-board head-dipping, novel object exploration, open-field activity, startle, PPI) differentiate the strains at G3-G5 but not at G1, suggesting that the pre-/post-natal environment may have influenced these co-segregated traits at G1, though further selection pressure along the subsequent generations (G1-G5) rescues the typical strain-related differences.

Association between prepulse inhibition of the startle response and latent inhibition of two-way avoidance acquisition: A study with heterogeneous NIH-HS rats

This study presents the first evaluation of the associations between responses in two paradigms related to schizophrenia in the genetically heterogeneous NIH-HS rat stock. NIH-HS rats are a stock of genetically heterogeneous animals that have been derived from eight different inbred strains. A rotational breeding schedule has been followed for more than eighty generations, leading to a high level of genetic recombination that makes the NIH-HS rats a unique tool for studying the genetic basis of (biological, behavioral, disease-related) complex traits. Previous work has dealt with the characterization of coping styles, cognitive and anxiety/fear-related profiles of NIH-HS rats. In the present study we have completed their characterization in two behavioral models, prepulse inhibition (PPI) and latent inhibition (LI) of the two-way active avoidance response, that appear to be related to schizophrenia or to schizophrenia-relevant symptoms. We have found that these rats display PPI for each of the four prepulse intensities tested, allowing their stratification in high, medium and low PPI subgroups. When testing these three subgroups for LI of two-way active avoidance acquisition it has been observed that the LowPPI and MediumPPI subgroups present impaired LI, which, along with the fact that the HighPPI group presents significant LI, allows us to hypothesize that responses in these two paradigms are somehow related and that selection of NIH-HS rats for Low vs HighPPI could make a promising animal model for the study of clusters of schizophrenia-relevant symptoms and their underlying neurobiological mechanisms.

Neonatal handling decreases unconditioned anxiety, conditioned fear, and improves two-way avoidance acquisition: a study with the inbred Roman high (RHA-I)- and low-avoidance (RLA-I) rats of both sexes.

The present study evaluated the long-lasting effects of neonatal handling (NH; administered during the first 21 days of life) on unlearned and learned anxiety-related responses in inbred Roman High- (RHA-I) and Low-avoidance (RLA-I) rats. To this aim, untreated and neonatally-handled RHA-I and RLA-I rats of both sexes were tested in the following tests/tasks: a novel object exploration (NOE) test, the elevated zero maze (ZM) test, a “baseline acoustic startle” (BAS) test, a “context-conditioned fear” (CCF) test and the acquisition of two-way active—shuttle box—avoidance (SHAV). RLA-I rats showed higher unconditioned (novel object exploration test -“NOE”-, elevated zero maze test -“ZM”-, BAS), and conditioned (CCF, SHAV) anxiety. NH increased exploration of the novel object in the NOE test as well as exploration of the open sections of the ZM test in both rat strains and sexes, although the effects were relatively more marked in the (high anxious) RLA-I strain and in females. NH did not affect BAS, but reduced CCF in both strains and sexes, and improved shuttle box avoidance acquisition especially in RLA-I (and particularly in females) and in female RHA-I rats. These are completely novel findings, which indicate that even some genetically-based anxiety/fear-related phenotypes can be significantly modulated by previous environmental experiences such as the NH manipulation.

Coping style and stress hormone responses in genetically heterogeneous rats: Comparison with the Roman rat strains

The purpose of the present study was to evaluate for the first time the stress-induced hypothalamus–pituitary–adrenal (HPA), adrenocorticotropic hormone (ACTH), corticosterone and prolactin responses of the National Institutes of Health genetically heterogeneous rat stock (N/Nih-HS rats) in comparison with responses of the relatively high and low stress-prone Roman Low- (RLA-I) and High-Avoidance (RHA-I) rat strains. The same rats were also compared (experiment 1) with respect to their levels of unconditioned anxiety (elevated zero-maze test), novelty-induced exploratory behavior, conditioned fear and two-way active avoidance acquisition. In experiment 2, naive rats from these three strains/stocks were evaluated for “depressive-like” behavior in the forced swimming test. N/Nih-HS and RLA-I rats showed significantly higher post-stress ACTH, corticosterone and prolactin levels than RHA-I rats. N/Nih-HS rats also presented the highest context-conditioned freezing responses, extremely poor two-way avoidance acquisition and very low novelty-induced exploratory behavior. Experiment 2 showed that, compared to RHA-I rats, N/Nih-HS and RLA-I rats displayed significantly less struggling (escape-directed) and increased immobility responses in the forced swimming test. Factor analysis of data from experiment 1 showed associations among behavioral and hormonal responses, with a first factor comprising high loadings of elevated zero-maze variables and lower loadings of conditioned fear, two-way avoidance acquisition and hormonal measures, while a second factor mainly grouped conditioned fear and two-way avoidance acquisition with novelty-induced exploration and post-stress prolactin. Thus, regarding their anxiety/fearfulness, passive coping style, “depressive-like” and stress-induced hormonal responses the N/Nih-HS rats resemble the phenotype profiles of the relatively high-anxious and stress-prone RLA-I rat strain.

Endocrinological Differences between Hatano High- and Low-Avoidance Rats during Early Two-Way Avoidance Acquisition

Hatano high (HAA)- and low (LAA)-avoidance rats were selected from Sprague-Dawley rats genetically on the basis of their active avoidance behavior in a shuttle-box test. The purpose of this study was to investigate stress-related alterations of hormones corticotropin-releasing hormone (CRH), arginine-vasopressin (AVP), prolactin, and adrenocorticotropin (ACTH) in the brain and blood during early avoidance acquisition using two lines of Hatano rats. In paraventricular nucleus (PVN) of the hypothalamus, the CRH levels in HAA rats were significantly increased after shuttle-box tasks compared with before the tasks, whereas the CRH levels in LAA rats significantly decreased after shuttle-box tasks compared with before the tasks. In the HAA rats, the CRH and AVP levels in the median eminence decreased after shuttle-box tasks, whereas there were no significant differences in the levels between before and after shuttle-box tasks in LAA rats. The plasma concentrations of ACTH were significantly higher in HAA rats than in LAA rats after shuttle-box tasks. These results show that the response of CRH-ACTH was higher in HAA rats than in LAA rats. This phenotype may be an important reason for the high avoidance rates of shuttle-box tasks in HAA rats. These endocrine differences in early avoidance acquisition may be involved in regulation of their avoidance responses in the shuttle-box task.

Two-way avoidance acquisition is negatively related to conditioned freezing and positively associated with startle reactions: A dissection of anxiety and fear in genetically heterogeneous rats

The double, fear-driven "passive avoidance/active avoidance" conflict appearing during acquisition of two-way active avoidance, involves high levels of anxiety and a dominant tendency for freezing responses, which in turn run against the appearance of active escape/avoidance behavior. In the present study, by using a large sample genetically heterogeneous (N/Nih-HS) rats, we have tested the hypothesis that rats showing relatively higher levels of context-conditioned freezing (during the initial trials of that task) should show lower efficiency to acquire two-way avoidance behavior, i.e. the prediction that the initial context-conditioned freezing in the shuttle box would be negatively related to avoidance acquisition efficiency. In agreement with such a hypothesis, the results from the three rat subsamples used show that context-conditioned freezing (during the first 5 inter-trial intervals of the 40-trial two-way avoidance session) is negatively correlated (r=-0.34 to r=-0.64, p<0.001) with two-way avoidance acquisition, in a way that subgroups of rats with extremely high or low levels of freezing markedly differ in their avoidance performance: "high freezer" rats show much worse avoidance acquisition than "low freezers". Moreover, the relationships of conditioned freezing and avoidance acquisition with baseline and fear-potentiated startle, as well as with unconditioned anxiety (in the elevated zero-maze test), have also been studied. Taken collectively, the results indicate that: (i) context conditioned freezing is a reliable (and negative) predictor of two-way avoidance acquisition; (ii) baseline and fear-potentiated startle responses show positive associations with avoidance responding, and (iii) unconditioned anxiety in the elevated zero-maze is also negatively associated with two-way avoidance acquisition. Such patterns of associations are considered to be very informative in regard to the search for (common or differential) neural and genetic mechanisms of different forms of (unlearned or learned) anxious or fear responses.

Differential effects of cohort removal stress on the acoustic startle response of the Roman/Verh rat strains.

Male and female rats of the inbred Roman/Verh strains, which have been psychogenetically selected and bred for good (RHA-I/Verh) and extremely poor (RLA-I/Verh) two-way avoidance acquisition, were evaluated in an acoustic startle response test. One half of the rats of each strain and sex were previously subjected to 30 min of isolation by removal of their cage partners ("cohort removal"). During the testing session each animal received 40 acoustic stimuli at interstimulus intervals of 30 seconds. The hyperemotional RLA-I/Verh rats (especially the males) showed stronger acoustic startle responses than did their RHA-I/Verh counterparts. Startle amplitudes of the RLA-I/Verh males were further enhanced by the stress of 30 min isolation, whereas cohort removal did not significantly affect startle response amplitudes in RHA-I/Verh rats of either sex or in females of the RLA-I/Verh strain.

Plasma ACTH levels during early, two-way avoidance acquisition in high- and low-avoidance rats (Hatano strains).

Having successfully bred for high- and low-avoidance rats (HAA and LAA, respectively) on a shuttlebox task, we performed three experiments designed to identify factors which might be related to the phenotypic differences seen in avoidance behavior. In experiment 1, shuttlebox behavior was measured to determine whether the phenotypic difference was activity related. In terms of intertrial responses, there was no difference between HAA and LAA rats in locomotor activity during the conditioning process. Experiment 2 compared adrenal weights of HAA and LAA rats at 11 weeks of age. The observation that the adrenal glands were heavier in HAA than in LAA rats suggested that these strains might differ in aspects of endocrine response. In experiment 3, plasma levels of ACTH and corticosterone were determined during early escape/avoidance acquisition in the shuttlebox. Plasma levels of ACTH after the shuttlebox testing were higher in HAA than in LAA rats. There was no difference between the two strains in plasma levels of corticosterone after testing, possibly due to a ceiling effect. These results suggest that the phenotypic differences in the acquisition of avoidance behavior of HAA and LAA rats may be related to different endocrine responses, rather than to locomotor activity.

CS modality transfer of two-way avoidance in rats with central and basolateral amygdala lesions

Post-lesion acquisition of two-way avoidance and subsequent transfer to two warning signals (conditioned stimulus, CS) of different modality were investigated in 60 rats. In Experiment I the animals were originally trained with less salient (darkness) CS, then transferred to more salient compound (darkness and white noise), and finally to white noise CS. The opposite arrangement of the conditioned stimuli (CSi) during the subsequent stages was employed in Experiment II. In control animals, avoidance acquisition was faster and the intertrial responding (ITR) rate lower with the auditory than with the visual CS. Lesioned rats learned avoidance responses more slowly, independently of CS modality. The transfer to other CSi revealed dramatic between-group difference in the level and consistency of avoidance response, shuttle-box latencies and ITR rate. In control animals, transfer to more salient CSi enhanced avoidance performance, whereas change to less salient CS decreased it. Rather small changes in shuttle-box performance and consistency of avoidance response due to CS modality were seen in rats with the basolateral lesions. In contrast, central nucleus injury caused a strong deterioration in the avoidance transfer, especially when the visual CS followed the acoustic one. The results indicate differential involvement of the basolateral and central amygdala nuclei in stimulus-processing mechanisms of instrumental defensive behavior.

The acoustic startle response in inbred Roman high- and low-avoidance rats.

To investigate the emotional reactions of two rat strains selectively bred for good and poor two-way avoidance acquisition (RHA/Verh and RLA/Verh), male animals of both strains were tested in an acoustic startle response test. They received 40 acoustic stimuli followed by 10 electric foot shocks and another 30 acoustic stimuli. RLA/Verh rats showed a significantly higher startle response compared to RHA/Verh animals, indicating a stronger emotional reaction to acoustic stimuli. In addition, the former showed a stronger response to foot shocks. Combined with earlier findings, we conclude that selection for two-way avoidance learning does not result in cognitive defects in the RLA/Verh strain but, rather, in stronger emotional reactions to fearful stimuli.

Shuttlebox performance in BALB/cByJ, C57BL/6ByJ, and CXB recombinant inbred mice: Environmental and genetic determinants and constraints

Male mice from nine strains (the CXB recombinant inbred set and their progenitors, BALB/cByJ and C57BL/6ByJ) were tested in a shuttlebox for two-way avoidance acquisition in an effort to resolve discrepancies between earlier studies. Mice were tested at one of three times of day (TOD) using one of three apparatus configurations. Analysis of variance revealed complex interactions between strain, apparatus configuration, and TOD. Subsequent analyses and tests of simple effects indicated very strong influences of apparatus configuration and/or TOD upon rate of acquisition in some strains, but not in others. C57BL/6 mice were relatively little affected by either variable; BALB/c mice were responsive to apparatus configuration but not to TOD. The recombinant inbred strains performed variously at levels that were either intermediate to, comparable with, or higher than those of the progenitors, depending upon the context of the testing situation. Genetic determination of behavior is, in part, a function of the particular context in which the behavior occurs. Both TOD and apparatus configuration are significant environmental factors.

Differential interactions between ethanol and Ro 15-4513 on two anxiety tests in rats

The effects of low (2 g/kg) and high (4 g/kg) doses of ethanol and their interaction with the imidazobenzodiazepine Ro 15-4513 (a partial inverse agonist of the benzodiazepine receptor) were studied in two different models of anxiety in rats: the “elevated plus-maze” test and the early acquisition of two-way (shuttlebox) avoidance. In the elevated plus-maze, ethanol (2 g/kg) increased the percentage of entries into the open arms (%EOA) and both ethanol doses increased the percentage of time spent into the open arms (%TOA), thus indicating an anxiolytic action which was reversed by Ro 15-4513 (5 mg/kg). By contrast, Ro 15-4513 did not counteract the anxiolytic effect of the low dose of ethanol in the acquisition of shuttlebox avoidance. Thus, the treatment with 2 g/kg of ethanol (plus either vehicle or Ro 115-4513) significantly. increased the total number of avoidances. Conversely, animals treated with 4 g/kg of ethanol showed impaired shuttlebox avoidance acquisition, and this effect was completely reversed by Ro 15-4513. Ro 15-4513 was without effect on its own in any of the anxiety-related parameters (i.e., %EOA, %TOA, and avoidance acquisition). The results indicate a different pattern of ethanol effects and Ethanol × Ro 15-4513 interactions depending upon the task used.

The early acquisition of two-way (shuttle-box) avoidance as an anxiety-mediated behavior: Psychopharmacological validation

Several lines of evidence have established that performance during the initial steps of acquisition on a shuttle-box avoidance task is an anxiety-mediated behavior (i.e., the differences between strains selectively bred for emotionality; the effects of postnatal handling; the course of the corticosterone response and behavioral measures of fear during acquisition). The present study was carried out to add pharmacological evidence to that view by testing the action of anxiogenic and anxiolytic drugs. Single 40-trial sessions with mild shocks (0.4 mA–0.6 mA) were used. In the first experiments the action of sodium pentobarbital (1.25, 2.5 and 5 mg/kg) and three benzodiazepines (diazepam, 2 and 4 mg/kg; alprazolam, 1 1.25 and 1.5 mg/ kg and adinazolam, 1, 2, 4 and 6 mg/kg) were tested. The last two experiments tested a possible proanxiety action of Ro 15–4513 (2, 5 and 10 mg/kg) and FG 7142 (5, 10 and 15 mg/kg), two partial inverse agonists of benzodiazepine receptors, which previous data had suggested to be anxiogenic. The results showed that the measure of acquisition of a two-way active avoidance is a sensitive mean for detecting either anxiolytic or anxiogenic effects of drugs, independently of their effects on locomotor activity, thus suggesting that such test could be a valid model of anxiety in animals.

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.

Bidirectional avoidance by mice as a function of CS, US, and apparatus variables

Acquisition of two-way avoidance by mice was slower with a light CS than with a buzzer CS, with punishment of intertrial responses than without punishment, and with a short CS-CS interval than with a long CS-CS interval (30 vs. 60 sec). Light-cued avoidance was little affected by shock level (.35–1.5 mA), whereas mice trained with the buzzer CS learned faster at 1.5 mA. Animals required to move away from light or toward light showed comparable rates of acquisition. Other CS, US, and apparatus variables (directionality of cue, maximal shock duration, and presence vs. absence of a central partition in the shuttlebox) interacted in a complex fashion with those already mentioned. This resulted in widely differing performances in what may superficially appear to be different versions of the same task. The differences in mouse and rat responses to some of the variables can contribute to an understanding of the interactions between organismic and test factors and the relative explanatory value of alternative avoidance models.

Amphetamine, conditioned stimulus, and nondebilitating preshock effects on activity and avoidance: Further evidence for interactions between associative and nonassociative changes

A literature survey and preliminary experiments with rats on the consequences of shock preexposure on subsequent activity and escape or avoidance showed the need for further work on the interactions between nondebilitating preshock and various test and treatment factors. The two main experiments used 16 preexposure conditions, namely, presence or absence of unavoidable punishment (36 shocks of 2.5 mA and 5 sec subdivided in three daily sessions), a light CS, a central partition in the shuttle-box, and dl-amphetamine sulfate (1 mg/kg ip 15 min before each session). In both experiments the four factors studied exerted more than additive effects on activity in preexposure sessions, leading to a very high frequency of crossing in the CS-shock-no-partition-drug condition. Upon retesting for activity (Experiment 1) suppression of locomotion by prior shock was less marked in animals preexposed to CS-US pairings in the absence of partition, while proactive amphetamine effects consisted mainly of a progressive increase of activity over successive retest sessions in the groups not preshocked. Upon retesting for light-cued, two-way avoidance acquisition (Experiment 2) the groups preexposed to US only were mostly retarded, while those preexposed to paired CS and US were mostly facilitated. Other changes, including drug pretreatment consequences, were negligible or unsystematic, but in general the data showed that the effects of various preexposure conditions on activity could not account for those on avoidance. Overall, it appears that the interactions between nondebilitating preshock and other test and treatment factors can be further exploited to clarify the respective roles of various associative and nonassociative mechanisms in modulation of activity and adaptive responding in aversive situations.

Effects of paradoxical sleep deprivation on two-way avoidance acquisition

Immediately following 72 hrs of paradoxical sleep (PS) deprivation, Wistar rats were given a shuttle-box avoidance task consisting of 50 trials. At an interval of 6 days retention was assessed through a reacquisition session of 20 trials. In a first experiment, the pendulum technique was employed for PS deprivation and its effects were compared with those produced by the conventional watertank technique. The first technique consists of arousing animals from sleep before PS can arise, by swinging their cages in a way that produces postural imbalance at regular intervals. A moving pendulum just not causing imbalance in the animals served as control for this technique. As control for the watertank technique the large platform was chosen. Shuttle-box avoidance was greatly impaired during the second half of the acquisition session in both platform conditions as compared to both pendulum conditions. A relatively small deficit was found toward the end of the session in both PS deprivation conditions as compared to both control conditions. No differences in avoidance were established during retention testing, suggesting that performance rather than learning deficits occurred during acquisition. In a second experiment, the multiple platform was used for PS deprivation. This modified version of the watertank technique also disrupted performance during shuttle-box avoidance acquisition. However, this effect appeared to be less pronounced than the effect found in the classical platform condition of Experiment 1. It is concluded that the performance deficit induced by both the classical and the multiple platform condition was mainly due to nonspecific effects.

Inescapable shock alters mescaline's disruption of active avoidance acquisition.

Rats were tested on a two-way avoidance acquisition with or without inescapable shock given 24 h prior to training. Mescaline given to nonshock rats disrupted acquisition in a dose-dependent fashion and tolerance developed to this disruption. Mescaline given to shock rats had no effect on acquisition even though levels of acquisition were the same for both shock and nonshock rats without drug. Moreover, subchronic treatment (5 days facilitated acquisition. These experiments demonstrate an interaction between shock, which presumably is a stressor, and mescaline. The data are consistent with the observation that when animals are exposed to presumptive stressors (e.g., shock, handling) hallucinogens can facilitate behavior, while in other situations, hallucinogens disrupt behavior.

Evidence for a role of the locus coeruleus noradrenaline system in learning

The effect of the new noradrenaline (NA) neurotoxin, DSP4, on the acquisition of one-way and two-way active avoidance responses was studied in rats. This treatment caused a marked degeneration of the iocus coeruleus NA system and markedly impaired both one- and two-way avoidance acquisition. Both effects were blocked by pretreatment with desipramine, a NA uptake blocking agent. The present findings support the hypothesis that the locus coeruleus NA system in its entire extent may play a role in aversive learning.

Maternal behavior in two rat lines selected for differences in the acquisition of two-way avoidance.

The maternal behavior of Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats was studied, using a time-sampling method. It was concluded that: a)RLA/Verh mothers spent more time with their young, b) RHA/Verh mothers were more active, and c) the mothers of both lines mostly "blanketed' their young during nursing, although the RHA/Verh mothers assumed the side-nursing position more often than their counterparts.