Brain c-Fos immunocytochemistry and cytochrome oxidase histochemistry after a fear conditioning task (original) (raw)
Related papers
Distinct patterns of brain Fos expression in Carioca High- and Low-conditioned Freezing Rats
PLOS ONE, 2020
Background The bidirectional selection of high and low anxiety-like behavior is a valuable tool for understanding the neurocircuits that are responsible for anxiety disorders. Our group developed two breeding lines of rats, known as Carioca High-and Low-conditioned Freezing (CHF and CLF), based on defensive freezing in the contextual fear conditioning paradigm. A random selected line was employed as a control (CTL) comparison group for both CHF and CLF lines of animals. The present study performed Fos immunochemistry to investigate changes in neural activity in different brain structures among CHF and CLF rats when they were exposed to contextual cues that were previously associated with footshock. Results The study indicated that CHF rats expressed high Fos expression in the locus coeruleus, periventricular nucleus of the hypothalamus (PVN), and lateral portion of the septal area and low Fos expression in the medial portion of the septal area, dentate gyrus, and prelimbic cortex (PL) compared to CTL animals. CLF rats exhibited a decrease in Fos expression in the PVN, PL, and basolateral nucleus of the amygdala and increase in the cingulate and perirhinal cortices compared to CTL animals. Conclusions Both CHF and CLF rats displayed Fos expression changes key regions of the anxiety brain circuitry. The two bidirectional lines exhibit different pattern of neural activation and inhibition with opposing influences on the PVN, the main structure involved in regulating the hypothalamic-pituitary-adrenal neuroendocrine responses observed in anxiety disorders.
Brain Research, 2008
Rats selectively bred for high or low levels of emotionality represent an important and powerful tool to investigate the role of genetic variables in the occurrence of different anxiety disorders. In the present study, albino rats were selectively bred for differences in defensive freezing behavior in response to contextual cues previously associated with footshock, an animal model of general anxiety disorder. The results indicate that these two new lines of rats, which we refer to as Carioca High-Freezing (CHF) and Carioca Low-Freezing (CLF), show a reliable difference in conditioned freezing after three generations of selection.
Behavioral Neuroscience, 2004
. The effects of contextual fear conditioning on the release of acetylcholine ACh in the hippocampus of freely moving rats was Ž assessed using microdialysis. Measures were carried out during both acquisition and retention testing re-exposure to the conditioning . Ž . chamber and compared between animals that either received foot-shocks as unconditioned stimulus conditioned group or no Ž . foot-shocks control group during acquisition. Results showed that during acquisition, hippocampal ACh extracellular level was increased with respect to baseline but that this increase was of similar magnitude in both groups. By contrast, re-exposure to the Ž . conditioning chamber the day after retention testing produced a significantly greater increase in ACh extracellular level in the Ž . Ž conditioned that, otherwise, displayed conditioned freezing behavior to contextual cues , than in the control group which displayed . virtually no freezing . This enhanced hippocampal ACh release seems to result from the greater hippocampal processing of contextual stimuli in conditioned animals with respect to controls. q
Expression of c-Fos and CRF in the brains of rats differing in the strength of a fear response
Behavioural brain …, 2008
The aim of the study was to examine the neurochemical background of differences in the individual responses to conditioned aversive stimuli, using the strength of a rat conditioned freezing response (the contextual fear test), as a discriminating variable. It was shown that low responders (LR), i.e. rats with duration of a freezing response one standard error, or more, below the mean value, had a higher activity of the M2 cortical area, and the median raphe nucleus (c-Fox expression), in comparison to the high responders (HR), i.e. rats with the duration of a freezing response one standard error, or more, above the mean value. These animals had also stronger 5-HT-and CRF-related immunostaining in the M2 area, and increased concentration of GABA in the basolateral nucleus of amygdala (in vivo microdialysis). The LR group vocalized more during test session in the aversive band, and had higher serum levels of corticosterone, examined 10 min after test session. It was shown that different natural patterns of responding to conditioned aversive stimuli are associated with different involvement of brain structures and with dissimilar neurochemical mechanisms.
Cued Fear Conditioning in Carioca High- and Low-Conditioned Freezing Rats
Frontiers in Behavioral Neuroscience, 2020
Anxiety disorders (AD) comprise a broad range of psychiatric conditions, including general anxiety (GAD) and specific phobias. For the last decades, the use of animal models of anxiety has offered important insights into the understanding of the association between these psychopathologies. Here, we investigate whether Carioca high-and low-conditioned freezing rats (CHF and CLF, respectively), a GAD animal model of anxiety, show similar high-and low-freezing behavioral phenotypes for cued auditory fear conditioning. Adult CHF (n = 16), CLF (n = 16) and normal age-matched Wistar rats (control, CTL, n = 16) were tested in a classical auditory-cued fear conditioning paradigm over 3 days (Tone + Shock and Tone only groups, n = 8 per treatment). Freezing responses were measured and used as evidence of fear conditioning. Overall, both CHF and CLF rats, as well as CTL animals displayed fear conditioning to the auditory CS. However, CLF animals showed a rapid extinction to the auditory conditioned stimulus compared to CHF and CTL rats. We discuss these findings in the context of the behavioral and neuronal differences observed in rodent lines of high and low anxiety traits.
Brain research, 2002
Several sources of evidence indicate that the inferior colliculus also integrates acoustic information of an aversive nature besides its well-known role as a relay station for auditory pathways. Gradual increases of the electrical stimulation of this structure cause in a hierarchical manner alertness, freezing and escape behaviors. Independent groups of animals implanted with bipolar electrodes into the inferior colliculus received electrical stimulation at one of these aversive thresholds. Control animals were submitted to the same procedure but no current was applied. Next, analysis of Fos protein expression was used to map brain areas activated by the inferior colliculus stimulation at each aversive threshold and in the controls. Whereas alertness elicited by stimulation of the inferior colliculus did not cause any significant labeling in any structure studied in relation to the respective control, electrical stimulation applied at the freezing threshold increased Fos-like immunoreactivity in the central amygdaloid nucleus and entorhinal cortex. In contrast, escape response enhanced Fos-like immunoreactivity in the nucleus cuneiform and the dorsal periaqueductal gray matter of the mesencephalon. This evidence supports the notion that freezing and escape behaviors induced by electrical stimulation of the inferior colliculus activate different neural circuitries in the brain. Both defensive behaviors caused significant expression of c-fos in the frontal cortex, hippocampus and basolateral amygdaloid nucleus. This indistinct pattern of c-fos distribution may indicate a more general role for these structures in the modulation of fear-related behaviors. Therefore, the present data bring support to the notion that amygdala, dorsal hippocampus, entorhinal cortex, frontal cortex, dorsal periaqueductal gray matter and cuneiform nucleus altogether play a role in the integration of aversive states generated at the level of the inferior colliculus.
Experimental Neurology, 2007
The inferior colliculus (IC) is an important relay station for ascending auditory information to the medial geniculate nucleus (MGN) and temporal cortex. It has been reported that the ventral (ICv) and dorsal (ICd) regions of the IC are involved with the defensive reaction and audiogenic seizures, respectively. As freezing is the first response induced by stimulation of these IC nuclei with increasing doses of N-methyl-Daspartate (NMDA), a question that arises is whether or not fear and audiogenic seizures generated at the IC level are interrelated processes. To address this issue, the Fos distribution in selected limbic structures following injections of NMDA into the ICv or ICd at freezing (7 nmol)-and escape (20 nmol)-producing doses was examined. Freezing behavior induced by intra-ICd NMDA caused an increase of Fos expression in the MGN, superior colliculus, dorsal columns of the periaqueductal gray and locus coeruleus while freezing induced by intra-ICv NMDA caused a significant Fos immunoreactivity in the prelimbic (PrL) and cingulate (Cg) cortices, basolateral and medial nuclei of the amygdala, ventrolateral periaqueductal gray, cuneiform nucleus and locus coeruleus. Escape behavior induced by NMDA injections into both nuclei caused a widespread Fos labeling in all limbic structures examined in this study. These results suggest that distinct circuits underlie the freezing behavior generated at the level of ICd and ICv. This is the first study to map Fos distribution associated with the stimulation of the ICv and ICd, regions supposed to be involved with fear and audiogenic seizures, respectively.
Disrupting basolateral amygdala function impairs unconditioned freezing and avoidance in rats
European Journal of Neuroscience, 2001
Lesions of the lateral/basolateral amygdala nuclei (BLC) disrupt freezing behaviour in response to explicit or contextual cues (conditioned stimuli ± CS) paired previously with footshock (unconditioned stimulus). This de®cit in expression of defensive behaviour in response to conditioned stimuli is often interpreted as inability of lesioned rats to learn CS±US associations. However, ®ndings of several studies indicate that BLC-lesioned rats can rapidly learn CS±US associations. Such ®ndings suggest that lesioned rats can learn CS±US associations but are impaired in the expression of freezing behaviour. In the present study we report that both temporary inactivation (lidocaine) and permanent excitotoxic (NMDA) lesions of the BLC impair the unconditioned freezing and avoidance behaviours of rats in response to a novel fear-eliciting stimulus, a ball of cat hair. These ®ndings suggest that the BLC in¯uences the expression of freezing and avoidance behaviours, and/or that it potentiates rats' experience of fear. Along with prior evidence of spared memory for aversive learning after BLC lesions, these ®ndings suggest that disrupted freezing to conditioned cues in BLC-lesioned rats does not necessarily re¯ect inability to form CS±US associations.
After extinction of conditioned fear, memory for the conditioning and extinction experiences becomes context dependent. Fear is suppressed in the extinction context, but renews in other contexts. This study characterizes the neural circuitry underlying the context-dependent retrieval of extinguished fear memories using c-Fos immunohistochemistry. After fear conditioning and extinction to an auditory conditioned stimulus (CS), rats were presented with the extinguished CS in either the extinction context or a second context, and then sacrificed. Presentation of the CS in the extinction context yielded low levels of conditioned freezing and induced c-Fos expression in the infralimbic division of the medial prefrontal cortex, the intercalated nuclei of the amygdala, and the dentate gyrus (DG). In contrast, presentation of the CS outside of the extinction context yielded high levels of conditioned freezing and induced c-Fos expression in the prelimbic division of the medial prefrontal cortex, the lateral and basolateral nuclei of the amygdala, and the medial division of the central nucleus of the amygdala. Hippocampal areas CA1 and CA3 exhibited c-Fos expression when the CS was presented in either context. These data suggest that the context specificity of extinction is mediated by prefrontal modulation of amygdala activity, and that the hippocampus has a fundamental role in contextual memory retrieval.