Tomasz Werka

Functional anatomy of neural circuits regulating fear and extinction

The memory of fear extinction is context dependent: fear that is suppressed in one context readily renews in another. Understanding of the underlying neuronal circuits is, therefore, of considerable clinical relevance for anxiety disorders. Prefrontal cortical and hippocampal inputs to the amygdala have recently been shown to regulate the retrieval of fear memories, but the cellular organization of these projections remains unclear. By using anterograde tracing in a transgenic rat in which neurons express a dendritically-targeted PSD-95:Venus fusion protein under the control of a c-fos promoter, we found that, during the retrieval of extinction memory, the dominant input to active neurons in the lateral amygdala was from the infralimbic cortex, whereas the retrieval of fear memory was associated with greater hippocampal and prelimbic inputs. This pattern of retrieval-related afferent input was absent in the central nucleus of the amygdala. Our data show functional anatomy of neural circuits regulating fear and extinction, providing a framework for therapeutic manipulations of these circuits.

Social modulation of learning in rats

It is well known that emotions participate in the regulation of social behaviors and that the emotion displayed by a conspecific influences the behavior of other animals. In its simplest form, empathy can be characterized as the capacity to be affected by and/or share the emotional state of another. However, to date, relatively little is known about the mechanisms by which animals that are not in direct danger share emotions. In the present study, we used a model of between-subject transfer of fear to characterize the social interaction during which fear is transmitted, as well as the behavioral effects of socially transmitted fear. We found that (1) during social interaction with a recently fear-conditioned partner, observers and demonstrators exhibit social exploratory behaviors rather than aggressive behaviors; (2) learning and memory in a shock-motivated shuttle avoidance task are facilitated in rats that underwent a social interaction with a partner that had been fear conditioned; and (3) a brief social interaction with a recently fear-conditioned partner immediately before fear conditioning increases conditioned freezing measured on the next day. The observed effects were not due to a stress-induced increase in pain sensitivity or analgesia. Collectively, these data suggest that a brief social interaction with a cage mate that has undergone an aversive learning experience promotes aversive learning in an otherwise naïve animal. We argue that socially transferred fear is an adaptation that promotes defensive behavior to potentially dangerous situations in the environment.

C-fos protooncogene expression in rat brain after long-term training of two-way active avoidance reaction

C-fos nuclear protooncogene encodes a regulatory protein (Fos), able to directly influence both expression of itself and other genes. It has been repeatedly shown that c-fos expression coincides with different forms of cell activation, probably being functionally involved in the coupling of extracellular ligands to long-term cellular responses. In this study it has been found that c-fos mRNA accumulation in rat brain, as measured by northern blotting coincides with increase of performance level of learned behavior of a two-way active avoidance task. We have previously reported (Nikolaev et al., Brain Res. Bull., in press) that a single training session of two-way active avoidance strongly induces c-fos mRNA accumulation but that after long-term training up to the asymptotic level of performance no c-fos expression was detectable. In this paper we show that c-fos still remains inducible even after long-term, asymptotic training to darkness as conditioned stimulus (CS), provided that a novel stimulus, wide band noise, which elevated performance level, was given together with darkness as compound CS.

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.

Social modulation in extinction of aversive memories.

Return of fear after extinction is a considerable challenge for the efficacy of exposure-based therapies. Fear recovery is most often modeled in the laboratory by changing the experimental context and studied in isolated animals. Since social context is an important factor affecting behavior, the question arises how it influences the recovery of extinguished fear. Here we present two novel behavioral models that allow studying social modulation of extinction memory retrieval. We show that the presence of a fearful cage mate results in a robust renewal of freezing as well as avoidance responses that were previously successfully extinguished.

Sex differences in social modulation of learning in rats.

In its simplest form, empathy can be characterized as the capacity to share the emotional experiences among individuals, a phenomenon known as emotional contagion. Recent research shows that emotional contagion and its adaptive role can be studied in rodents. However, it is not known whether sex differences observed in human empathy extend to its more primitive forms. In the present study, we used a rat model of emotional contagion to compare the behavioral consequences of social transfer of information about threat, and the subsequent neural activation patterns in male and female rats. We found that: (1) males and females display a similar behavioral pattern during the interaction with either a fear-conditioned or a control rat; (2) interaction with a fear-conditioned conspecific positively modulates two-way avoidance learning in male and diestral female rats but not in estral females; and (3) such interaction results in increased c-Fos expression in the central and lateral nuclei of the amygdala and the prelimbic and infralimbic cortex in males, whereas in females no such changes were observed. Collectively, our results point to the occurrence of sex and estrus cycle phase differences in susceptibility to emotional contagion and underlying neuronal activation in rodents.

Inhibition of delay of the two-way avoidance response and warning-signal salience

Effects of extended training (nine sessions, 50 trials each) on two-way avoidance response latencies were studied. For each rat, auditory and visual warning signals (CS) were presented on separate trials, either according to a Gellermann series or in 25-trial blocks. Intermittent presentation of the two kinds of trials yielded shorter latencies and higher levels of avoidance performance in response to the noise CS than in response to the darkness CS. Presentation of trials in blocks revealed stronger response-eliciting properties of stimuli presented during the second half of the session than of those presented during the first half. A significant decrease of avoidance performance in the early portions of the delay period, an index of inhibition of delay, was obtained on auditory trials presented in the second half of the sessions. Results indicate that strong fear of the warning signal is required for inhibition of delay of avoidance responses.

Effects of partial lesion of dorsal hippocampal afferent and GM1 ganglioside treatment on conditioned emotional response and hippocampal afferent markers in rats

Acquisition of the conditioned emotional response (CER) in 32 male hooded rats previously learned to press a bar for food and divided into four groups was studied. Two groups received electrolytic lesions of the dorsal hippocampal afferent and were thereafter injected either with GM1 ganglioside (30 mg/kg daily) or with buffer. Two remaining groups were sham operated and similarly injected. The partial hippocampal deafferentation evoked immediate enhancement of bar presses rate which persisted during the 2-week period of testing. CER training undertaken 2 days after surgical procedures appeared unsuccessful, whereas similar training with a cue of different modality initiated a week later resulted in acquisition of conditioned suppression of bar presses in all groups. Toward the end of training the conditioned suppression was more pronounced in lesioned than in control rats. The GM1 injections attenuated the conditioned suppression in control rats, presumably due to an antinociceptive role of ganglioside treatment. Behavioural training did not change the normal distribution pattern in cholinergic and serotonergic hippocampal afferent markers showing dorso-ventral gradient along longitudinal axis. The lesion-induced decrease pattern was also not affected. However, in contrast to previous findings in non-trained animals, the GM1 treatment was not effective in protecting against degenerative changes in the hippocampus of trained rats.

Effects of stimulus modality on the shuttle activity in rats.

Shuttle activity during repeated presentation of irrelevant auditory (white band noise) and visual (darkness) stimuli was studied in 32 male Möll-Wistar rats. The subjects were randomly divided into two groups. In Group ND the auditory stimulus was used in the first habituation session, and the visual stimulus in the second habituation session. The opposite sequence of stimuli was applied in Group DN. House light was used as a background stimulus in both groups of subjects. The rate of crossing from one compartment to the other was markedly enhanced by noise, regardless of the position of the auditory stimulus in the sessions sequence. A clear decrease of response rate was caused by noise termination, whereas neither darkness offset nor onset changed the relatively high and stable rate of responding. The results of this study provided convincing evidence of the energizing influence of the irrelevant auditory cue on the shuttle activity in rats. Moreover, these findings showed that a habituation procedure provides an efficient tool to analyze pure unconditional features of the to-be-conditioned stimuli, and to foresee several response modifications during further conditioning.