Subdiaphragmatic vagotomy does not attenuate c-Fos induction in the nucleus of the solitary tract after conditioned taste aversion expression

After acquisition of a conditioned taste aversion (CTA) against sucrose, intraoral infusions of sucrose induce c-Fos-like immunoreactivity (c-FLI) in the medial intermediate nucleus of the solitary tract (iNTS) of the rat. In order to determine if c-FLI expression in the iNTS depends on subdiaphragmatic vagal afferent input to the NTS secondary to gastrointestinal symptoms during CTA expression (e.g. diarrhea), we quantified the induction of c-FLI in the iNTS by sucrose infusions after total subdiaphragmatic vagotomy in rats with a previously acquired CTA against sucrose. Rats were conditioned against intraoral infusions of sucrose by pairing sucrose infusions with toxic LiCl injections. After CTA acquisition, rats underwent bilateral subdiaphragmatic vagotomy or were sham-vagotomized. One week after surgery, rats received an intraoral infusion of sucrose. One hour after the test infusion, rats were perfused and processed for c-FLI. Vagotomy had no apparent effect on the behavioral expression of the previously acquired CTA, because both vagotomized and sham-vagotomized rats rejected all of the test intraoral infusion of sucrose. There was also no significant difference between vagotomized and sham-vagotomized rats in the number of c-FLI-positive cells in the iNTS after CTA expression. We conclude that c-FLI induction correlated with CTA expression is not dependent on subdiaphragmatic vagal efferent output or afferent input.     

Brain mechanisms of taste aversion learning in the rat.

This study aims to reveal brain mechanisms underlying the conditioned taste aversion (CTA) learning. To establish CTA in Wistar male adult rats, 0.01 M Na-saccharin and IP injection of 0.15 M LiCl were used for conditioned stimulus and unconditioned stimulus, respectively. Rats with ibotenic acid lesions of the pontine taste area, thalamic taste area, or basolateral nucleus of the amygdala, failed to establish CTA learning, but lesions of the amygdaloid nuclei other than the basolateral nucleus, cortical gustatory area, hippocampus, entorhinal cortex, bed nucleus of the stria terminalis, or substantia innominata, showed slight or little effects. Rats that received amino-phosphovaleric acid chronically in the amygdala failed to establish CTA. These results, together with our preliminary results, suggest that long-term potentiation of gustatory responses involving N-methyl-D-aspartate receptors in the basolateral nucleus of the amygdala is a basic mechanism for CTA learning.     

Phosphorylation of MAP kinase and CREB in mouse cortex and amygdala during taste aversion learning

Mice were subjected to a taste aversion conditioning procedure in which they drank water, familiar saccharin, or novel saccharin, followed by injection of either NaCl or the emetic agent LiCl. Immunohistochemistry was used to localize phosphorylated MAP kinase (ppERK) and phosphoCREB (pCREB) in the brain shortly after conditioning. An examination of insular cortex and amygdala revealed specific phosphorylation of MAP kinase by novel saccharin and LiCl, and CREB by LiCl. Pairing of novel saccharin with LiCl was the only stimulus sufficient to increase ppERK and pCREB immunoreactivity in the lateral amygdala, suggesting this as a site of CS-US convergence. ppERK immunoreactivity was localized to both nuclear and non-nuclear compartments, suggesting multiple functional roles of MAP kinase during learning.     

Effects of insular cortex lesions on conditioned taste aversion and latent inhibition in the rat

The present study tested the hypothesis that lesions of the insular cortex of the rat retard the acquisition of conditioned taste aversions (CTAs) because of an impairment in the detection of the novelty of taste stimuli. Demonstrating the expected latent inhibition effect, nonlesioned control subjects acquired CTAs more rapidly when the conditioned stimulus (0.15% sodium saccharin) was novel rather than familiar (achieved by pre-exposure to the to-be-conditioned taste cue). However, rats with insular cortex lesions acquired taste aversions at the same slow rate regardless of whether the saccharin was novel or familiar. The pattern of behavioural deficits obtained cannot be interpreted as disruptions of taste detection or stimulus intensity, but is consistent with the view that insular cortex lesions disrupt taste neophobia, a dysfunction that consequently retards CTA acquisition because of a latent inhibition-like effect.     

Insular cortex lesions alter conditioned taste avoidance in rats differentially when using two methods of sucrose delivery

The insular gustatory cortex may be essential for the evaluation of saliency and representation of the incentive values of tastes. Gustatory cortex lesions should interfere with conditioned taste avoidance according to these factors, which depend on the conditioned taste avoidance protocol used. The present study was aimed at investigating the effects of bilateral lesions of the gustatory cortex-focusing on electrolytic and excitotoxic lesions. Lesioned and sham-operated male Long-Evans rats were intoxicated using LiCl after drinking sucrose from a tube (SD) or having the same amount of sucrose fed directly into their mouths through a chronically implanted intra-oral (IO) cannula. Every aspect of the experiment was carefully counterbalanced between the experimental groups. In the control groups, the acquired avoidance towards sucrose was strongly preserved over eight extinction test days in SD rats but not in IO rats, in which a progressive decline was recorded. Electrolytic gustatory cortex lesions impaired but did not suppress conditioned taste avoidance in both protocols. Excitotoxic lesions tend to impair CTA also, but differentially according to the SD or IO protocols. Extinction of CTA was selectively impaired in the SD protocol by small lesions destroying the anterior insular cortex.     

Differential involvement of gustatory insular cortex and amygdala in the acquisition and retrieval of conditioned taste aversion in rats.

Lesion studies of the role of the gustatory insular cortex (GC) and amygdala (Am) in conditioned taste aversion (CTA) are confounded by the irreversibility of the intervention. Functional ablation methods allow more specific influencing of different phases of CTA acquisition and retrieval. Bilateral tetrodotoxin (TTX) blockade of GC (10 ng) or Am (3 ng) before or after saccharin drinking in rats with chronically implanted intracerebral cannulae showed that GC is indispensable for the initial processing of the taste stimulus but not for the association of the gustatory trace with the symptoms of LiCl poisoning. Gustatory signals can by-pass the blocked Am, the inactivation of which, however, impairs the gustatory trace-poisoning association. TTX injection into both GC and Am impairs CTA retrieval more than isolated blockade of either of these structures. It is argued that GC and Am implement processing of gustatory and visceral signals, respectively, but that formation and consolidation of the CTA engram proceeds outside forebrain, probably at the level of the brainstem.     

Induction of a brainstem correlate of conditioned taste aversion expression: role of the pontine parabrachial nucleus

Increases in Fos-like immunoreactivity (FLI) in the intermediate division of the nucleus of the solitary tract (iNTS) are seen following the expression of a conditioned taste aversion (CTA). In studies limited to behavioral assessment, the pontine parabrachial nucleus (PBN) has been demonstrated to play a critical role in the acquisition, but not the expression, of CTAs. To better define the role of the PBN in taste aversion learning, the present study examined the effects of PBN lesions on FLI in iNTS in animals with lesions placed either before or after CTA training. As is the case with behavioral expression of a CTA, timing of PBN lesions was found to be critical. Lesions placed prior to conditioning blocked evidence of conditioning, including both taste rejection and FLI in iNTS. Lesions placed after conditioning, but before testing, did not interfere with either taste rejection or FLI. These results support and extend prior claims that PBN is critical for CTA acquisition but not expression. They also demonstrate that input from PBN to iNTS is not necessary for the FLI seen there during CTA expression.     

Effects of anterior basolateral amygdala lesions on taste aversions produced by high and low oral doses of LiCl and lactose in the rat

Adult male hooded rats (n = 12) with bilateral electrolytic lesions centered on the anterior basolateral amygdala (BLA) were given training using procedures meant to produce two different types of conditioned taste aversions (CTAs) to test whether the disruptive effects of such lesions on this form of learning were dependent upon the dosages of the illness-inducing agents used as unconditioned stimuli (UCSs). The CTAs were produced by either LiCl-induced toxicosis or lactose malabsorption. Comparison with sham-operated control subjects (n = 8) indicated that the disruptive effects of the lesions were inversely related to the dosage level of LiCl but not lactose: whilst the lesions disrupted CTAs produced by a low i.p. dose of LiCl and by a low, but not a high, oral dose of LiCl, they did not alter CTA learning produced by either high or low oral doses of lactose. These results were interpreted as providing further evidence that BLA mediates only certain types of CTA learning.     

Insular cortex lesions and taste aversion learning: effects of conditioning method and timing of lesion

The specific role of insular cortex in acquisition and expression of a conditioned taste aversion was assessed using two different conditioning methods, which vary mode of taste delivery. Involvement of insular cortex in the induction of c-Fos-immunoreactivity in the nucleus of the solitary tract, a cellular correlate of the behavioral expression of a conditioned taste aversion, was also assessed. Electrolytic lesions of insular cortex blocked behavioral expression of a conditioned taste aversion and this was evident not only when lesions were placed prior to conditioning, but also when they were made after conditioning but before testing. In contrast to the effects on behavior, lesions did not completely block the c-Fos-immunoreactivity which accompanies re-exposure to the aversive taste. In addition, the blocking of behavioral evidence of aversion conditioning by cortical lesions was seen both in animals trained under an intraoral acquisition procedure and those trained with bottle-conditioning. This contrasts with previous work with amygdala lesions which showed that amygdala was absolutely necessary for taste aversions conditioned with the intraoral method but not for those conditioned using bottle presentation of the taste. Overall, these findings imply that the details of the neural circuitry involved in taste aversion learning, including its anatomical distribution, complexity and degree of redundancy, vary with the type of conditioning method employed.     

c-Fos-like immunoreactivity in the brainstem following gastric loads of various chemical solutions in rats

The distribution of c-Fos-like immunoreactivity (c-FLI) in the lower brainstem especially in the area postrema (AP), nucleus of the tractus solitarius (NTS) and parabrachial nucleus (PBN) was examined following gastric loads of various chemical solutions in rats. An aliquot of 7.5 ml of each stimulus was intragastrically infused, and c-FLI was detected. The most remarkable c-FLI was induced by LiCl, lactose and ethanol which are known to be effective unconditioned stimuli in conditioned taste aversions. Polycose and disaccharides such as sucrose and maltose induced more c-FLI than monosaccharides such as glucose, fructose and galactose. Relatively low levels of c-FLI were observed for other sweeteners such as saccharin, glycine and alanine, and other basic taste stimuli such as NaCl, HCl, quinine and umami substances. Each stimulus induced a similar proportion of c-FLI among the subnuclei of the NTS, but not in the PBN, where chemicals effective in inducing conditioned taste aversions elicited stronger c-FLI in the external lateral subnucleus, and those in inducing conditioned taste preferences such as Polycose and glucose elicited stronger c-FLI in the dorsal lateral subnucleus. Vagotomy reduced c-FLI to about 50% for LiCl stimulation and to about 30% for sucrose stimulation, suggesting that LiCl has a larger proportion of extravagal inputs than sucrose.     

Insular cortex and amygdala lesions differentially affect acquisition on inhibitory avoidance and conditioned taste aversion

These experiments examined the effects of NMDA-induced lesions of the amygdala and insular (gustatory) cortex (IC) on inhibitory avoidance learning and conditioned taste aversion (CTA) in rats. IC lesions, but not amygdala lesions, disrupted CTA. In contrast, lesions of either brain region disrupted inhibitory avoidance learning. These findings support the view that the IC is strongly involved in the acquisition of external as well as visceral aversively motivated behavior. Despite extensive functional interconnections, these 2 brain regions appear to have different roles in mediating different forms of aversively based learning.     

cFos induction during conditioned taste aversion expression varies with aversion strength

Fos-like immunoreactivity (FLI) can indicate the location of neurons activated following expression of conditioned taste aversion (CTA). After one conditioning trial FLI has been identified in the intermediate nucleus of the solitary tract (iNTS) with little expression in other brain regions. The present study assessed the effect of increasing aversion strength on the magnitude and anatomical distribution of FLI during CTA expression. When animals received three rather than one conditioning trial, significant FLI was seen not only in the iNTS but also in the parabrachial nucleus (PBN), and the central nucleus of the amygdala (CNA), regions thought to be important in taste aversion learning.     

Behavioral changes and expression of heat shock protein hsp-70 mRNA, brain-derived neurotrophic factor mRNA, and cyclooxygenase-2 mRNA in rat brain following seizures induced by systemic administration of kainic acid

Kainic acid-induced seizures in rats represent an established animal model for human temporal lobe epilepsy. However, it is well-known that behavioral responses to the systemic administration of kainic acid are inconsistent between animals. In this study, we examined the relationship between expression of genes, neuropathological damage, and behavioral changes (seizure intensity and body temperature) in rats after systemic administration of kainic acid. The considerable differences in the response to kainic acid-induced seizures were observed in rats after a single administration of kainic acid (12 mg/kg i.p.). There was no detection of the expression of heat shock protein hsp-70 mRNA and HSP-70 protein in brain of vehicle-treated controls and in animals exhibiting weak behavioral changes (stage 1–2). A moderate expression of hsp-70 mRNA was detected throughout all regions (the pyramidal cell layers of CA1–3 and dentate gyrus) of the hippocampus, the basolateral, lateral, central and medial amygdala, the piriform cortex, and the central medial thalamic nucleus of rats that developed moderate seizures (stage 3–4). Marked expression of hsp-70 mRNA was detected in the all regions (cingulate, parietal, somatosensory, insular, entorhinal, piriform cortices) of cerebral cortex and all regions of hippocampus, and the central medial thalamic nucleus of the rats that developed severe seizures (stage 4–5). In addition, marked HSP-70 immunoreactivity was detected in the pyramidal cell layers of CA1 and CA3 regions of hippocampus, all regions (cingulate, parietal, somatosensory, insular, piriform cortices) of cerebral cortex, and the striatum of rats that developed severe seizures (stage 4–5). Furthermore, a marked expression of cyclooxygenase-2 (COX-2) mRNA and brain-derived neurotrophic factor (BDNF) mRNA levels by kainic acid-induced behavioral seizures (stage 3–4 or stage 4–5) was detected in all hippocampal pyramidal cell layers, granule layers of dentate gyrus, piriform cortex, neocortex, and amygdala. The present study suggest that the behavioral changes (seizure intensity and body temperature) and neuropathological damage after systemic administration of kainic acid are inconsistent between animals, and that these behavioral changes (severity of kainic acid-induced limbic seizures) might be correlated with gene expression of hsp-70 mRNA, COX-2 mRNA, and BDNF mRNA in rat brain.     

Establishing aversive, but not safe, taste memories requires lateralized pontine-cortical connections

Aversive and safe taste memory processing is dramatically disrupted by bilateral lesions of the pontine parabrachial nucleus (PBN). To determine how such lesions affect patterns of neuronal activation in forebrain, lesions were combined with assessment of cFos-like immunoreactivity (FLI) in insular cortex (IC) and amygdala after conditioned taste aversion (CTA) training. Increases in FLI in amygdala and IC, which are normally seen following novel (versus familiar) CS-US pairing, were eliminated after PBN lesions. This suggests that PBN lesions prevent transmission of critical CS and US information to forebrain regions for the processing of both aversive and safe taste memories. Unilateral asymmetrical lesions of PBN and IC blocked CTA acquisition as well as normal patterns of FLI in amygdala after novel CS-US pairing, an effect not seen when unilateral lesions were confined to a single hemisphere. The crossed-disconnection experiments provide compelling evidence that functional interactions between PBN and IC are required for CTA acquisition, but not for safe taste memory formation and retrieval. The dissociation between effects of the different types of lesions on safe and aversive taste memories supports emerging evidence that the neural underpinnings of the two types of taste learning differ.     

Differential roles of the prefrontal cortical subregions and basolateral amygdala in compulsive cocaine seeking and relapse after voluntary abstinence in rats

Compulsive drug use and a persistent vulnerability to relapse are key features of addiction. Imaging studies have suggested that these features may result from deficits in prefrontal cortical structure and function, and thereby impaired top‐down inhibitory control over limbic–striatal mechanisms of drug‐seeking behaviour. We tested the hypothesis that selective damage to distinct subregions of the prefrontal cortex, or to the amygdala, after a short history of cocaine taking would: (i) result in compulsive cocaine seeking at a time when it would not usually be displayed; or (ii) facilitate relapse to drug seeking after abstinence. Rats with selective, bilateral excitotoxic lesions of the basolateral amygdala or anterior cingulate, prelimbic, infralimbic, orbitofrontal or anterior insular cortices were trained to self‐administer cocaine under a seeking–taking chained schedule. Intermittent mild footshock punishment of the cocaine‐seeking response was then introduced. No prefrontal cortical lesion affected the ability of rats to withhold their seeking responses. However, rats with lesions to the basolateral amygdala increased their cocaine‐seeking responses under punishment and were impaired in their acquisition of conditioned fear. Following a 7‐day abstinence period, rats were re‐exposed to the drug‐seeking environment for assessment of relapse in the absence of punishment or cocaine. Rats with prelimbic cortex lesions showed decreased seeking responses during relapse, whereas those with anterior insular cortex lesions showed an increase. Combined, these results show that acute impairment of prefrontal cortical function does not result in compulsive cocaine seeking after a short history of self‐administering cocaine, but further implicates subregions of the prefrontal cortex in relapse.     

Amygdala-gustatory insular cortex connections and taste neophobia

To examine whether communication between the amygdala and gustatory insular cortex (GC) is required for normal performance of taste neophobia, three experiments were conducted. In Experiment 1, rats with asymmetric unilateral lesions of the basolateral amygdala (BLA) and the GC displayed elevated intake of a novel saccharin solution relative to control subjects. However, an attenuation of neophobia was not found following asymmetric unilateral lesions of the GC and medial amygdala (MeA; Experiment 2) or of the MeA and BLA (Experiment 3). This pattern of results indicates that the BLA and GC functionally interact during expression of taste neophobia and that the MeA functionally interacts with neither the BLA nor the GC. Research is needed to further characterize the nature of the involvement of the MeA in taste neophobia and to determine the function of the BLA-GC interaction during exposure to a new taste.     

Differential effects of neurotoxin-induced lesions of the basolateral amygdala and central nucleus of the amygdala on lithium-induced conditioned disgust reactions and conditioned taste avoidance

When rats are intraorally exposed to saccharin solution that has previously been paired with lithium chloride (LiCl), they display Pavlovian conditioned disgust reactions. When exposed to LiCl-paired saccharin solution by bottle, they display suppressed instrumental approach to the bottle resulting in suppressed consumption. The present experiments demonstrated that while neither neurotoxin-induced lesions of the basolateral amygdala (BLA) nor the central nucleus of the amygdala (CeA) attenuated the display of Pavlovian conditioned disgust reactions, lesions of the BLA (but not the CeA) attenuated instrumental conditioned avoidance of the taste. The results are discussed in light of current models of the role of the amygdala in aversive learning.     

Glutamate in the parabrachial nucleus of rats during conditioned taste aversion

Brain microdialysis combined with HPLC and spectroscopic detection was used to monitor extracellular glutamate in the parabrachial nucleus (PBN) of rats during acquisition of a conditioned taste aversion (CTA). Microdialysis fractions taken every 20 min were used to assess the effects of presentation of the conditioned stimulus alone (CS, consumption of 0.1% saccharin), the unconditioned stimulus alone (US, intraperitoneal injection of 0.15 M LiCl, 2% b.w. induced malaise after water drinking) as well as that of CS-US pairing. After 15 min of saccharin drinking, the glutamate concentration in the eluate (20 microl/20 min) reached 80% above the baseline but returned to the basal value in the next fraction. LiCl alone (applied 1 h after 15 min drinking of water) increased glutamate only following some delay, i.e. in the second and third post-lithium fraction by 90 and 67%, respectively. However, when LiCl was injected 1 h after the onset of saccharin intake, the glutamate concentration rose significantly (by 95%) already in the first post-LiCl fraction and by 120% in the second one. It appears, therefore, that the 'saccharin trace' facilitates the effect of lithium on extracellular concentration of glutamate in PBN during acquisition of CTA.     

Brain-derived neurotrophic factor enhances conditioned taste aversion retention

Brain-derived neurotrophic factor (BDNF) has recently emerged as one of the most potent molecular mediators of not only central synaptic plasticity, but also behavioral interactions between an organism and its environment. Our previous studies on the insular cortex (IC), a region of the temporal cortex implicated in the acquisition and storage of conditioned taste aversion (CTA), have demonstrated that induction of long-term potentiation (LTP) in the projection from the basolateral nucleus of the amygdala (Bla) to the IC, previous to CTA training, enhances the retention of this task. Recently, we found that intracortical microinfusion of BDNF induces a lasting potentiation of synaptic efficacy in the Bla-IC projection of adult rats in vivo. In this work, we present experimental data showing that intracortical microinfusion of BDNF previous to CTA training enhances the retention of this task. These findings support the concept that BDNF may contribute to memory-related functions performed by a neocortical area, playing a critical role in long-term synaptic plasticity.     

Effects of conditioned taste aversion on extracellular serotonin in the lateral hypothalamus and hippocampus of freely moving rats

This study used microdialysis to monitor extracellular levels of 5-HT and its metabolite, 5-hydroxyindole acetic acid (5-HIAA) in the lateral hypothalamus (LH) and hippocampus of freely moving rats that had developed a CTA to a 2.5 mM saccharin solution (CS) following its pairing with illness induced by lithium chloride (US). Results showed that oral infusion of the saccharin CS significantly enhanced extracellular LH 5-HT in animals that had developed a taste aversion compared with control groups, including unconditioned (CS-no US) and pseudoconditioned (no CS-US) subjects. As an anatomical control, the hippocampus was identified based on previous research suggesting that it is not integrally involved in CTA learning or retrieval and that 5-HT in this brain site does not directly mediate feeding behavior but is closely correlated with arousal. In contrast with the results obtained in the LH, hippocampal 5-HT was not preferentially elevated in subjects in the CTA group but rather was increased to the same extend in both CTA and control groups after saccharin infusion. Moreover, the increase in LH 5-HT for the CTA group was nearly twice that observed in the hippocampus for any group. Acute administration of LiCl elevated extracellular 5-HT to similar levels in both sites, well above the changes observed following conditioning. 5-HIAA was unaffected in either brain site by oral infusion of saccharin solution or injection of LiCl.(ABSTRACT TRUNCATED AT 250 WORDS)