Odor-taste pairings lead to the acquisition of negative hedonic qualities by the odor in aversion learning.

Three experiments examined the affective responses conditioned to an odorous stimulus in the taste-mediated odor aversion learning paradigm. Experiment 1 analyzed the microstructure of licking behavior during voluntary consumption. Before conditioning, water-deprived rats had access to a bottle containing either a tasteless odor (0.01% amyl acetate) diluted in water or mixed with 0.05% saccharin. Next, the rats were injected with either LiCl or saline immediately after drinking saccharin. At test, they received the odor and taste solutions on separate days. Lick cluster size was used as a direct measure of the hedonic response to the odor cue. Rats receiving odor-taste pairings prior to the saccharin devaluation showed both lower consumption and lick cluster size, reflecting a reduced hedonic evaluation of the odor. Experiments 2a and 2b used the orofacial reactivity method. After pretraining in the drinking boxes with the odor alone or mixed with saccharin, the rats were intraorally infused with saccharin before injection with LiCl or saline. At test, they were infused in separate sessions with the odor and taste and their orofacial reactions video recorded. There were increased aversive orofacial responses to the odor in rats that had prior odor-taste experience, a result indicating a negative hedonic evaluation of the odor. These results provide evidence of conditioned changes in affective value of odor cues through taste-mediated learning and are consistent with the idea that odor-taste pairings lead to the acquisition of taste qualities by the odor.

Characterisation of the neural basis underlying appetitive extinction & renewal in Cacna1c rats.

Recent studies have revealed impairments in Cacna1c ± heterozygous animals (a gene that encodes the Cav 1.2 L-type voltage-gated calcium channels and is implicated in risk for multiple neuropsychiatric disorders) in aversive forms of learning, such as latent inhibition, reversal learning or context discrimination. However, the role of Cav 1.2 L-type voltage-gated calcium channels in extinction of appetitive associations remains under-investigated. Here, we used an appetitive Pavlovian conditioning task and evaluated extinction learning (EL) with a change of context from that of training and test (ABA) and without such a change (AAA) in Cacna1c ± male rats versus their wild-type (WT) littermates. In addition, we used fluorescence in situ hybridization of somatic immediate early genes (IEGs) Arc and Homer1a expression to scrutinize associated changes in the medial prefrontal cortex and the amygdala. Cacna1c ± animals successfully adapt their responses by engaging in appetitive EL and renewal. However, the regional IEG expression profile changed. For the EL occurring in the same context, Cacna1c ± animals presented higher IEG expression in the infralimbic cortex and the central amygdala than controls. The prelimbic region presented a larger neural ensemble in Cacna1c ± than WT animals, co-labelled for the time window of EL in the original context and prolonged exposure to the unrewarded context. With a context change, the Cacna1c ± infralimbic region displayed higher IEG expression during renewal than controls. Taken together, our findings provide novel evidence of distinct brain activation patterns occurring in Cacna1c ± rats after appetitive extinction and renewal despite preserved behavioral responses. This article is part of the Special Issue on "L-type calcium channel mechanisms in neuropsychiatric disorders".

Characterizing Hedonic Responses to Flavors Paired with Internal Pain and Nausea through the Taste Reactivity Test in Rats.

Feeding behavior is a complex experience that involves not only sensory (i.e., visual, odor, taste, or texture) but also affective or emotional aspects (i.e., pleasure, palatability, or hedonic value) of foods. As such, behavioral tests that assess the hedonic impact of foods are necessary to fully understand the factors involved in ingestive behavior. In this protocol, we use the taste reactivity (TR) test to characterize the hedonic responses of rats to flavors paired with either lithium chloride-induced nausea or internal pain produced by hypertonic NaCl, two treatments that reduce voluntary consumption. This application of the TR test demonstrates how emetic and non-emetic (somatic pain in particular) treatments produce dissociable patterns of hedonic reactions to fluids: only emetic treatments result in the production of aversive orofacial responses, reflecting conditioned nausea, whereas somatic pain produces immobility, reflecting conditioned fear. Other methods, such as the microstructural analysis of licking behavior, do not reliably distinguish conditioned nausea and fear, a key advantage of the more selective TR procedure. This protocol also contains guidance for adaptation to other species and designs.

Socially conditioned flavor preferences with fluids: Transfer with solid foods, palatability, and testing constraints.

In two experiments, the social influence on choosing between two novel diets was analyzed. In Experiment 1, a group of rats (demonstrators) ingested a novel flavor cue presented in powdered food or in a 4% sucrose solution. Afterwards, demonstrators interacted with a group of observer rats for 30 minutes. Preferences for the cue flavor and another flavor were tested in observers using a two-choice test in the same or in the opposite substrate (solid/liquid) as their demonstrators. When tested with solid foods, observers displayed higher intake of the flavor consumed by the demonstrators, regardless of whether the demonstrators had consumed it as a solid or liquid. However, when observers were tested with solutions, there was no demonstration of the same preference. Experiment 2 focused on solutions, presenting them at test either with the flavor as a water solution alone, or as a solution plus an aerosol presentation. In addition, a single-cue testing procedure was used to allow examination of the palatability (assessed through the analysis of licking behavior) of the test solutions. Under single-cue testing procedures with solutions, observer rats did consume more of the flavors previously consumed by the demonstrator animals, but there was no effect of social learning on the palatability of the test flavors. These results suggest that socially conditioned flavor preferences can be reliably observed with fluid solutions, will transfer between different substrates, and affect consumption to a greater degree than palatability. However, future experiments need to be performed to confirm some of these suggestions.

Conditioned hedonic responses elicited by contextual cues paired with nausea or with internal pain.

Pairing a taste with either internal pain or nausea, despite equivalent effects on voluntary consumption, has dissociable effects on hedonic responses: Only pairing with nausea results in the production of disgust reactions, while pairing with internal pain results in conditioned fear as indicated by immobility. Here, we use orofacial reactions to examine the hedonic responses elicited by contextual, nonflavor, cues paired with nausea produced by injection of lithium chloride (LiCl) or internal pain caused by injection of hypertonic saline. In Experiment 1, aversive orofacial responses were the specific context-elicited behaviors in the rats injected with LiCl, whereas immobility was seen in the animals injected with hypertonic saline. In Experiment 2, rats first received discriminative training with two contexts, where one context was paired with LiCl or hypertonic saline, and the other context with isotonic saline. After this, rats were intraorally infused with a flavor (conditioned stimulus (CS) +) in the paired context, and with a different flavor (CS-) in the unpaired context. Second-order conditioning was then examined in a test conducted in the unpaired context. The infusion of the CS + flavor produced aversive orofacial responses in the rats injected with LiCl but immobility in the subjects injected with hypertonic saline. The results suggest that nonflavor cues support conditioned hedonic responses in the same way as flavor cues, which implies that the quality of aversion learning (conditioned nausea vs. fear) is primarily determined by the nature of the aversive event and not the type of conditioned cue. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Partial reinforcement and conditioned taste aversion: No evidence for resistance to extinction.

The partial reinforcement extinction effect (PREE) is the observation that, following training in which a response is followed by reward on only a subset of trials, the response is more resistant to extinction following the total removal of reward than it is after training in which reward is presented on all trials. The PREE is almost ubiquitous in instrumental conditioning procedures but only inconsistently observed in Pavlovian conditioning. In his classic review of animal learning, Mackintosh attributes the bulk of the PREE to generalisation decrement relating to the fact that partial reinforcement typically ensures that acquisition of responding has taken place in conditions similar to that of extinction (e.g., in the absence of the reinforcer). We report here that extinction of a conditioned taste aversion is not retarded by partial reinforcement in terms of either consumption of the taste or hedonic reactions to it (assessed through the analysis of licking microstructure). These results are consistent with Mackintosh's analysis of the PREE and the way in which it might differ between instrumental and Pavlovian conditioning.

Reward Adaptation and the Mechanisms of Learning: Contrast Changes Reward Value in Rats and Drives Learning.

Formal theories of learning suggest that associations between events are determined by the internal representations of those events. Thus, learning should depend on perceived reward value-even when perceptions differ from objective values. We examined this prediction in flavor-preference learning in rats. In two experiments, simultaneous contrast either increased perceived reward value, which was paired with a distinctive flavor cue (the positive conditioned stimulus, CS+), or decreased the perceived value of the same reward, which was then paired with a second flavor (the negative conditioned stimulus, CS-). Even though the CS+ and CS- were paired with the same objective reward, there was a preference for the CS+ in subsequent tests. Moreover, the size of contrast-produced changes in reward value during training predicted the preference for the CS+ at test. This contrast-produced learning effect illustrates the mechanisms by which associations, which normally track veridical relationships between events in the world, are formed.

Flavors paired with internal pain or with nausea elicit divergent types of hedonic responses.

Pairing a taste with either internal pain (e.g., from hypertonic saline injection) or nausea (e.g., from LiCl administration) will reduce subsequent consumption of that taste. Here we examine the responses to a taste paired with either hypertonic saline or LiCl using the analysis of licking microstructure (mean lick cluster size: Experiments 1-3), taste reactivity (examining the distribution of appetitive and aversive orofacial responses: Experiments 2-3), and immobility (as a measure of fear: Experiments 2-3). At both high (10 ml/kg 0.15 M LiCl, 10 ml/kg 1.5 M NaCl) and low dose levels (2 ml/kg 0.15 M LiCl, 4 ml/kg 1.5 M NaCl), pairing a taste with either LiCl-induced nausea or internal pain produced by hypertonic NaCl caused reductions in voluntary consumption, in appetitive taste reactivity responses, and in lick cluster size. However, only pairing with LiCl resulted in conditioned aversive taste reactivity responses to the taste. In contrast, pairing with hypertonic NaCl resulted in the taste eliciting higher levels of immobility (reflecting fear) than did pairing the taste with LiCl. The clearly dissociable effects of LiCl and hypertonic saline on aversive taste reactivity and fear responses, despite equivalent effects on consumption, demonstrates selective conditioning effects between internal pain and nausea. (PsycINFO Database Record

Blocking of flavor-nausea learning by non-flavor cues: Assessment through orofacial reactivity responses.

We investigated, using orofacial reactivity assessment, whether nonflavor context cues can elicit conditioned aversive reactions, and also whether context cues interfere, through blocking, with the reduction in taste palatability during taste aversion conditioning. Experiment 1 showed that a context previously paired with LiCl evoked aversive orofacial reactions, and also attenuated the reduction in palatability of a saccharin solution which was paired with LiCl in that context. In Experiment 2, this blocking effect was abolished when the rats were given nonreinforced exposure to the previously LiCl-paired context (context extinction) before aversive conditioning of the saccharin in compound with the context. These results confirm that context stimuli can elicit conditioned aversive reactions in the absence of any flavor component, and demonstrate that context cues can interfere with the affective aspects of taste aversion learning. Thus nonflavor cues appear to engage the same processes as taste cues in aversion learning. These results are consistent with the idea that taste aversion learning is governed by general associative mechanisms and the special properties of nausea, rather than by a selective mechanism for poison-avoidance. (PsycINFO Database Record

c-Fos activity in the insular cortex, nucleus accumbens and basolateral amygdala following the intraperitoneal injection of saccharin and lithium chloride.

This study examined c-Fos expression in selected brain areas consequent to intraperitoneal (IP) administration of saccharin and lithium chloride. Rats were tested for aversion to the saccharin as measured by flavor consumption and orofacial reactions in the taste reactivity (TR) test. It was found that intraperitoneal conditioning resulted in the reduction in voluntary consumption but not in the production of aversive orofacial responses to the saccharin. The immunohistochemistry quantification revealed increased c-Fos activity in the insular cortex, the shell and core regions of the nucleus accumbens, and the basolateral nucleus of the amygdala. These results show that a conditioned taste aversion can be induced without direct oropharyngeal gustatory stimulation at the time of conditioning. In addition, this study provide evidence of increased neural activity in response to intraperitoneal saccharin injections.

Functional brain networks underlying latent inhibition of conditioned disgust in rats.

The present experiment examined the neuronal networks involved in the latent inhibition of conditioned disgust by measuring brain oxidative metabolism. Rats were given nonreinforced intraoral (IO) exposure to saccharin (exposed groups) or water (non-exposed groups) followed by a conditioning trial in which the animals received an infusion of saccharin paired (or unpaired) with LiCl. On testing, taste reactivity responses displayed by the rats during the infusion of the saccharin were examined. Behavioral data showed that preexposure to saccharin attenuated the development of LiCl-induced conditioned disgust reactions, indicating that the effects of taste aversion on hedonic taste reactivity had been reduced. With respect to cumulative oxidative metabolic activity across the whole study period, the parabrachial nucleus was the only single region examined which showed differential activity between groups which received saccharin-LiCl pairings with and without prior non-reinforced saccharin exposure, suggesting a key role in the effects of latent inhibition of taste aversion learning. In addition, many functional connections between brain regions were revealed through correlational analysis of metabolic activity, in particular an accumbens-amygdala interaction that may be involved in both positive and negative hedonic responses.

Nonreinforced flavor exposure attenuates the effects of conditioned taste aversion on both flavor consumption and cue palatability.

Nonreinforced exposure to a cue tends to attenuate subsequent conditioning with that cue-an effect referred to as latent inhibition (LI). In the two experiments reported here, we examined LI effects in the context of conditioned taste aversion by examining both the amount of consumption and the microstructure of the consummatory behavior (in terms of the mean size of lick clusters). The latter measure can be taken to reflect affective responses to, or the palatability of, the solution being consumed. In both experiments, exposure to a to-be-conditioned flavor prior to pairing the flavor with nausea produced by lithium chloride attenuated both the reduction in consumption and the reduction in lick cluster sizes typically produced by taste aversion learning. In addition, we observed a tendency (especially in the lick cluster measure) for nonreinforced exposure to reduce neophobic responses to the test flavors. Taken together, these results reinforce the suggestion from previous experiments using taste reactivity methods that LI attenuates the effects of taste aversion on both consumption and cue palatability. The present results also support the suggestion that the failure in previous studies to see concurrent LI effects on consumption and palatability was due to a context specificity produced by the oral taste infusion methods required for taste reactivity analyses. Finally, the fact that the pattern of extinction of conditioned changes in consumption and in lick cluster sizes was not affected by preexposure to the cue flavors suggests that LI influenced the quantity but not the quality of conditioned taste aversion.

Latent inhibition of conditioned disgust reactions in rats.

The present experiments, using the latent inhibition (LI) paradigm, evaluated the effect of nonreinforced exposure to saccharin on the acquisition of an LiCl-induced saccharin aversion as measured by conditioned disgust reactions in the taste reactivity test and conditioned taste avoidance in a consumption test. When rats were preexposed to saccharin by bottle exposure (Experiments 1 and 3), LI was evidenced only by conditioned taste avoidance (bottle testing), but not by conditioned disgust reactions (intraoral [IO] testing). On the other hand, when rats were preexposed to saccharin by IO infusion (Experiments 2 and 3), LI was evidenced only by conditioned disgust reactions, but not by conditioned taste avoidance. Experiment 4 showed that LI of conditioned disgust reactions does not appear to be affected by a context shift from preexposure to testing phases. These results show that the expression of LI of both conditioned taste avoidance and conditioned disgust reactions depends critically on a common method of flavor exposure during preexposure and testing.