Conditioned Flavor Preferences Research Articles

Extinction of conditioned flavor preferences.

Although noted as a proponent of associative learning theory, Bob Rescorla acknowledged that other mechanisms might be responsible for the within-event learning produced when two stimuli co-occur. To investigate this possibility, he conducted experiments in which rats experienced a compound of a novel flavor and a palatable nutrient, and demonstrated that a preference for the flavor established by this training did not show the pattern of extinction that might be expected of a preference based on a flavor-nutrient association. A review is presented of subsequent work on the extinction of such conditioned flavor preferences in rats. The results are found to depend on the motivational state of the rat in training and on test, on the match between the procedures on training and test, and on the details of the test procedure (the nature of the choice offered to the rat). When conditions are arranged appropriately, the extinction effect (a loss of the conditioned response) expected by standard associative theory can be obtained. What remains a problem for this theory is the observation (made originally by Rescorla himself) is that the effects of extinguishing a conditioned flavor preference are remarkably persistent. The failure to obtain recovery from the effects of the extinction procedure remains as a signal that this form of learning may involve processes other than the association formation used to explain many other forms of learning. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Enteroendocrine cell types that drive food reward and aversion.

Animals must learn through experience which foods are nutritious and should be consumed, and which are toxic and should be avoided. Enteroendocrine cells (EECs) are the principal chemosensors in the GI tract, but investigation of their role in behavior has been limited by the difficulty of selectively targeting these cells in vivo. Here, we describe an intersectional genetic approach for manipulating EEC subtypes in behaving mice. We show that multiple EEC subtypes inhibit food intake but have different effects on learning. Conditioned flavor preference is driven by release of cholecystokinin whereas conditioned taste aversion is mediated by serotonin and substance P. These positive and negative valence signals are transmitted by vagal and spinal afferents, respectively. These findings establish a cellular basis for how chemosensing in the gut drives learning about food.

Rats learn to prefer the late-consumed flavor over the early-consumed flavor in a multi-flavored meal paired with oral glucose and corn oil

Conditioned flavor preference (CFP) is established by association: where a neutral flavor (conditioned stimulus, CS) is paired with orosensory and post-ingestive components of nutrients, including sugar and fat (unconditioned stimulus, US). A previous study reported that rats can learn to prefer flavors that they consumed earlier and later in a multi-flavored solution paired with an intragastric infusion of glucose, but they expressed only a preference for a late-consumed flavor when they were tested after feeding (Myers and Whitney, 2011). This paradigm can be a suitable rodent model to explain how humans acquire a selective preference for routinely late-served “dessert” foods and why these foods remain attractive even in the absence of hunger. Here, we examined whether oral glucose (Experiment 1) or fat (Experiment 2) acts as a US for flavor preference learning processes in this paradigm. In Experiment 1, adult female rats under food restriction were trained in 16 daily sessions with two distinct flavor CSs in succession per session; eight CS(+) sessions in which two distinct flavor CSs (early(+), late(+)) were sequentially presented for 8 min each with oral glucose (12%) as a US, and eight CS(-) sessions in which different CSs (early(-), late(-)) were unpaired with the US. In the 30-minute two-bottle choice test, rats preferred late(+) over late(-) only when tested 90 min after consumption of normal chow (fed test) but not after overnight deprivation (hungry test). Early(+) was not preferred over early(-) in both tests. Moreover, a significant preference for late(+) over early(+) was observed only in the fed test, which is a unique feature of oral glucose-CFP. These results indicate that taste sensations of oral glucose promote a rewarding effect of late-onset glucose nutrients. In Experiment 2, separate rats were trained with the same conditioning paradigm, but used a caloric matched fat solution (5.3% corn oil) for a US. The results showed that they expressed stronger preferences for early(+) and late(+) relative to their respective CS(-) flavors in both tests. Similar to Experiment 1, it was observed in the fed test that there was a preference for late(+) over early(+) in oral fat-CFP. Taken together, the present results suggest that routine timing arrangements can cause qualitative differences in conditioned preferences between multiple flavors within a sugar or fat-containing meal in rats, and that rats prefer the late-consumed flavor over the early-consumed flavor in the absence of hunger.

Asymmetric Control of Food Intake by Left and Right Vagal Sensory Neurons

Gut‐innervating vagal sensory neurons residing in the nodose ganglia (NG) conveys meal‐borne signals and control food intake, however, the underlying gut‐brain neurocircuit is unclear. The neuropeptide cocaine‐ and amphetamine‐regulated transcript (CART) is expressed in a subpopulation of vagal sensory neurons (~40%) that innervate the whole length of the gut and its release in the NTS is sufficient for meal termination. Thus, it can be an excellent target to understand the role of sensory neurons in feeding. Emerging evidence suggests that right or left NG (LNG or RNG) neurons recruit different downstream central circuits to control feeding, but the mechanisms remain poorly understood. Our hypothesis is that LNGCART and RNGCART neurons sense different meal‐related signals and are necessary and sufficient to modulate feeding.MethodsIn vivo calcium imaging, viral neuronal tracing, and feeding behaviors were assessed in response to chemo‐ and opto‐genetic stimulation of gut‐innervating vagal sensory neurons using CARTcre mice.ResultsAnatomical tracing revealed that LNGCART and RNGCART neurons innervate different regions of the duodenum (n=3). LNGCART innervate the muscular layer, while RNGCART innervate the villi. Importantly, 2‐photon calcium imaging highlighted real time increased activity in LNGCART in response to stretch, while intraduodenal fat infusions preferentially increased RNGCART neuron activity (n=7/group). Mice with LNGCART neurons ablation consumed higher volumes of liquid meal indicative of attenuated response to gastric and intestinal distension (n=6‐7/group). Furthermore, chemogenetic stimulation of LNGCART, but not RNGCART, caused pronounced and sustained hypophagia (n=7/group). Interestingly, mice with chemogenetic activation of RNGCART, but not LNGCART, increased flavor conditioning (n=7/group), while ablation of RNGCART neurons abolished the conditioned flavor preference that was paired with intragastric fat infusion (n=7/group) supporting their role in reward. In addition, optogenetic stimulation of RNGCART promoted self‐stimulation in a nose poke task (n=4). Lastly, dopaminergic neurons of the substantia nigra increased their firing rate in response to intraduodenal fat infusion, but not duodenal distension (n=2/group).ConclusionLeft and right vagus nerves provide different sensory information about a meal to the brain. Both anatomical and behavioral feeding studies support the necessity and sufficiency LNGCART neurons as food quantity detectors and RNGCART neurons as food reward mediators ‐ which has less influence on how much is consumed but rather plays a role in the decision‐making process about which foods to consume. Our findings on asymmetrical vagal innervation of the gut provides new mechanistic insight for how feeding decisions are made in response to vast and complex meal‐related signals.

The Effects of Roux-en-Y Gastric Bypass on Glucose- vs. Fructose-Associated Conditioned Flavor Preference

In rodents, repeated single-bottle exposures to distinctly flavored isocaloric glucose and fructose solutions, two sugars with different metabolic pathways, eventually lead to a preference for the former. Because Roux-en-Y gastric bypass (RYGB) surgery decreases preference for and intake of sugar solutions in rats, we tested whether RYGB would curtail the conditioning of a preference for a glucose-paired vs. fructose-paired flavor. RYGB (♂ n=11; ♀ n=10) and sham-operated (SHAM; ♂ n=9; ♀ n=10) rats were trained with a single bottle (30 min/day) containing 8% glucose solution flavored with either 0.05% grape or cherry Kool-Aid (Glu/CSG) or 8% fructose solution with the alternative Kool-Aid flavor (Fru/CSF) in an alternating fashion for 8 days. To determine baseline preferences, a 4-day 30-min two-bottle test was used to assess preference for Glu/CSG vs. Fru/CSF before training. After training, 2-day 30-min two-bottle tests assessed preference for the a) Glu/CSG (CSG-flavored 8% glucose solution) vs Fru/CSF (CSF-flavored 8% fructose solution), b) CSG- vs. CSF-flavored mixture of 4% glucose & 4% fructose (isocaloric), c) CSG- vs. CSF-flavored 0.2% saccharin (“sweet”, no calories), and d) CSG- vs. CSF-flavored water. During training, only male SHAM rats demonstrated progressively increased intake of Glu/CSG over Fru/CSF, and female SHAM rats displayed a trend. RYGB eliminated any difference in single-bottle intake of these solutions during training, regardless of sex. Like their male and female SHAM counterparts, male RYGB rats displayed a conditioned preference for the CSG-associated stimulus in Tests 1-3. Although female RYGB rats displayed acquisition of the conditioned flavor preference in Test 1, unlike the other groups, when the differential sugar cue between the two solutions was removed in Tests 2 and 3, female rats did not display a CSG preference. When the sugar and sweetener cues were both removed on Test 4, all groups displayed some generalization decrement. Thus, RYGB does not compromise the ability of rats to learn and express a glucose- vs. fructose-associated conditioned flavor preference when the exact CS used in training is presented in testing. The mechanistic basis for the sex difference in the effect of RYGB on the generalization decrement observed in this type of flavor preference learning warrants further study.

Caffeine-based flavor preference conditioning in the rat.

The massive consumption of caffeine-containing beverages has prompted many studies involving human participants that have obtained caffeine-based increases in liking for a flavor. However, few studies have succeeded in obtaining caffeine-based flavor preference learning in rats. The main aim of the present study was to examine the conditions under which such learning can be detected. Three experiments differed mainly in terms of the base solution to which caffeine was added. Using a base of maltodextrin and saccharin, Experiment 1 found modest increases in flavor preferences in both food- and fluid-restricted rats. Experiment 2 found a strong caffeine-based flavor preference when water, but not saccharin, was used as the base. Whereas the first two experiments used a within-subject design, in which one flavor was paired with caffeine and a second flavor was not, Experiment 3 used a between-subject design with fluid-restricted rats given almond-flavored water containing caffeine in the Paired condition but not in the Unpaired condition; caffeine-based flavor preference learning was again found. In Experiments 1 and 2 post-conditioning exposure to the flavor alone produced a decrease in preference. In summary, the main achievements of this study were to extend the conditions under which caffeine-based flavor preferences can be detected in rats and demonstrate that such learned preferences are subject to extinction.

Another look at the extinction of conditioned flavor preferences: Amount of training and tests for spontaneous recovery.

A conditioned flavor preference develops when hungry or thirsty rats experience a neutral flavor mixed in solution with a nutrient. In two sets of studies, we previously demonstrated that this learned preference is highly sensitive to flavor nonreinforcement (i.e., exposure to the flavor without the nutrient) either prior to (latent inhibition), during (partial reinforcement), or following (extinction) flavor-nutrient pairings. In each of these studies we employed a nutrient devaluation procedure to assess the integrity of specific flavor-nutrient associations following extinction, but more recently Gonzalez, Morillas, and Hall (Journal of Experimental Psychology: Animal Learning & Cognition, 42, 380-390, 2016) observed that sensitivity to extinction in thirsty rats in this preparation may depend upon use of a post-conditioning nutrient devaluation procedure. To assess the generality of our earlier results, but without including a post-conditioning nutrient devaluation phase, we assessed in three experiments the role of the number of flavor-nutrient pairings given prior to extinction and the possibility of spontaneous recovery following a 3-week delay. We observed that extinction consistently weakened the flavor preference in thirsty rats (in spite of the absence of a nutrient devaluation procedure) and also found no evidence for spontaneous recovery. These results establish that our prior findings that conditioned flavor preferences are weakened by extinction are quite robust in thirsty rats and that these extinction effects may be fairly permanent.

Sucrose-based flavor preferences in rats: Factors affecting detection of extinction.

Rats that have consumed a novel target flavor added to a sucrose solution will develop a preference for that flavor. Such preferences appear to persist over the course of many presentations of the flavor alone when animals are not food-deprived. However, previous research indicates that an extinction effect (a reduction in preference) can be obtained when training or testing is carried out in animals that are hungry. In a series of experiments that produced flavor preferences in hungry rats by adding the flavor to a sucrose solution, three (Experiments 1, 2A, 2B) established that the concentration of sucrose and the nature of the flavor influenced the results but failed to detect extinction. Two-bottle choice tests showed some loss of preference but this occurred both in subjects given the extinction treatment (flavor-only presentations) and in control subjects given just water. A loss of preference in rats given an extinction treatment as opposed to controls given only water was, however, found in Experiments 3 and 4. These experiments differed from Experiments 1 and 2 in that the extinction stage involved the presentation of two bottles containing the flavor, thus matching the two-bottle procedure used in the test phase. These results confirm that experiencing a flavor alone can result in extinction of a conditioned flavor preference in hungry rats but indicate that the effect is highly context-specific, requiring the conditions of the test to match those of the extinction procedure. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Nutrient-conditioned intake stimulation does not require a distinctive flavor cue in rats

The postoral actions of nutrients in rodents can stimulate intake and condition flavor preferences through an appetition process. Appetition is revealed in rodents by their increased intake of and preference for a flavored solution paired with intragastric (IG) nutrient infusions. Here we determined if IG 16% maltodextrin (MD) infusions can stimulate intake and preference in the absence of a distinctive flavor cue. Rats implanted with IG catheters were given chow and water 2 h/day followed, 2 h later, by 20-h oral access to water paired with IG MD infusions. Other rats were given bitter sucrose octaacetate solution (SOA) paired with IG MD infusions 20 h/day. Over 8 test days, the SOA rats increased their total 20-h fluid intake (oral + IG) from 26 to 119 g/20 h and Water rats increased their intake from 31 to 96 g/20 h. When infused IG with water instead of MD in a 4-day extinction test, the SOA and Water groups reduced their fluid intakes to 45–48 g/20 h. When oral fluids were again paired with IG MD infusions, the SOA and Water groups increased their intakes to 115 and 109 g/20 h, respectively. In two-bottle tests, the SOA rats drank more SOA paired with IG MD than water paired with IG water. Water rats given the choice of a water bottle paired with IG MD and water bottle paired with IG water did not consistently prefer the H2O/ID MD bottle. Instead they displayed side or sipper tube preferences although neither cue was consistently paired with IG MD during one-bottle training.

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.

Acquisition and expression of fat conditioned flavor preferences following dopamine D1, opioid and NMDA receptor antagonism in C57BL/6 mice

ABSTRACT Objectives: Inbred mouse strains differ in the pharmacology mediating sugar and fat intake and conditioned flavor preferences (CFP). C57BL/6, BALB/c and SWR inbred mice are differentially sensitive to dopamine (DA) D1, opioid and muscarinic receptor antagonism of sucrose, saccharin or fat intake, and to DA, opioid, muscarinic and N-methyl-D-aspartate (NMDA) receptor antagonism of acquisition of sucrose-CFP. DA D1, opioid and NMDA receptor antagonists differentially alter fat (Intralipid)-CFP in BALB/c and SWR mice. The present study examined whether naltrexone, SCH23390 or MK-801 altered acquisition and expression of Intralipid-CFP in C57BL/6 mice. Methods: In acquisition, groups of male food-restricted C57BL/6 mice received vehicle, naltrexone (1, 5 mg/kg), SCH23390 (50, 200 nmol/kg) or MK-801 (100, 200 μg/kg) before 10 training sessions in which mice alternately consumed two novel-flavored 5% (CS+) and 0.5% (CS-) Intralipid solutions. Six two-bottle CS choice tests followed with both flavors mixed in 0.5% Intralipid without injections. In expression, C57BL/6 mice underwent the 10 training sessions without injections followed by two-bottle CS choice tests 30 min following vehicle, naltrexone (1, 5 mg/kg), SCH23390 (200, 800 nmol/kg) or MK-801 (100, 200 μg/kg). Results: Fat-CFP acquisition in C57BL/6 mice was significantly though marginally reduced following naltrexone, SCH23390 and MK-801. Fat-CFP expression was similarly reduced by naltrexone, SCH23390 and MK-801 in C57BL/6 mice. Discussion: C57BL/6 mice were more sensitive to DA D1, opioid and NMDA antagonists in the expression of fat-CFP relative to sugar-CFP, but were less sensitive to DA D1 and NMDA antagonists in the acquisition of fat-CFP relative to sugar-CFP.

Hippocampal Input to the Nucleus Accumbens Shell Enhances Food Palatability

BackgroundInsight into the neural basis of hedonic processing has come from studies of food palatability in rodents. Pharmacological manipulations of the nucleus accumbens shell (NAcSh) have repeatedly been demonstrated to increase hedonic taste reactivity, yet the contribution of specific NAcSh circuit components is unknown. MethodsBidirectional optogenetic manipulations were targeted to the principal NAcSh projection neurons and afferent pathways in mice during free feeding assays. Number of licks per bout of consumption was used as a measure of food palatability as it was confirmed to track sucrose concentration and subjective flavor preferences. ResultsPhotoinhibition of NAcSh neurons, whether general or cell-type specific, was found to alter consumption without affecting its hedonic impact. Among the principal excitatory afferent pathways, we showed that ventral hippocampal (vHipp) input alone enhances palatability upon low-frequency photostimulation time-locked to consumption. This enhancement in palatability was independent of opioid signaling and not recapitulated by NAcSh or dopamine neuron photostimulation. We further demonstrated that vHipp input photostimulation is sufficient to condition a flavor preference, while its inhibition impedes sucrose-driven flavor preference conditioning. ConclusionsThese results demonstrate a novel contribution of vHipp–NAcSh pathway activity to palatability that may relate to its innervation of a particular region or neuronal ensemble in the NAcSh. These findings are consistent with the evidence that vHipp–NAcSh activity is relevant to the pathophysiology of anhedonia and depression as well as the increasing appreciation of hippocampal involvement in people’s food pleasantness ratings, hunger, and weight.

Forty-eight hour conditioning produces a robust long lasting flavor preference in rats

Conditioned flavor preference (CFP) learning is a form of associative learning in ingestive behavior. CFP Learning can be rapid and produces preferences of varying strengths that can be exceptionally persistent. We sought to establish a method to produce a robust long-lasting CFP in rats. Rats were given 48-h access (conditioning) to a CS+ flavor (grape or cherry 0.05% Kool-Aid, counterbalanced) mixed with 8% glucose and 0.05% saccharin. In order to determine the strength of conditioning rats were given 14 consecutive days of 24-h access to CS+ and CS- flavors mixed only with 0.05% Kool-Aid and 0.05% saccharin (extinction), then further tested 34 days after the last extinction test (48 days post conditioning) for 2 consecutive days with the CS+ and CS-. We found that not only did the learned CFP fail to extinguish over 14 days of testing, but it also persisted for at least 48 days after conditioning. These data provide a method to produce a robust, long lasting and persistent CFP for use in future ingestive behavior research.

Acquisition and expression of sucrose conditioned flavor preferences following dopamine D1, opioid and NMDA receptor antagonism in C57BL/6 mice

The study of inbred mouse strains is a useful animal model to assess differences in ingestive behavior responses, including conditioned flavor preferences (CFP). C57BL/6, BALB/c and SWR inbred mice display differential sensitivity to dopamine (DA) D1, opioid and muscarinic cholinergic receptor antagonism of sucrose or saccharin intake as well as to muscarinic cholinergic antagonism of acquisition (learning) of sucrose-CFP. Given that DA D1, opioid and N-methyl-D-aspartate (NMDA) receptor antagonists differentially alter sucrose-CFP in BALB/c and SWR inbred mice, the present study examined whether systemic administration of naltrexone, SCH23390 or MK-801 altered acquisition and expression of sucrose-CFP in C57BL/6 mice. In acquisition experiments, male food-restricted C57BL/6 mice were treated with vehicle, naltrexone (1, 5 mg/kg), SCH23390 (50, 200 nmol/kg) or MK-801 (100, 200 µg/kg) 30 min prior to each of ten daily sessions in which they alternately consumed a flavored (CS+, e.g. cherry) 16% sucrose solution and a differently-flavored (CS-, e.g. grape) 0.05% saccharin solution followed by six two-bottle CS choice tests mixed in 0.2% saccharin without injections. SCH23390 and MK-801, but not naltrexone eliminated sucrose-CFP acquisition in food-restricted C57BL/6 mice. In expression experiments, food-restricted C57BL/6 mice underwent the ten training sessions without injections followed by two-bottle CS choice tests 30 min following vehicle, naltrexone (1, 5 mg/kg), SCH23390 (200, 800 nmol/kg) or MK-801 (100, 200 µg/kg). SCH23390 more effectively reduced the magnitude of sucrose-CFP expression than naltrexone or MK-801 in food-restricted C57BL/6 mice. Thus, dopamine D1 and NMDA receptor signaling is essential for learning of sucrose-CFP in C57BL/6 mice.

Facilitation and retardation of flavor preference conditioning following prior exposure to the flavor conditioned stimulus.

In two experiments, rats received pairings of an almond flavor (Experiments 1 and 2B) or a vanilla flavor (Experiment 2A) with sucrose. In each experiment, half of the rats received prior exposure to the flavor and half were exposed to water. Conditioned preference was then assessed through two-bottle, flavor versus water, choice tests. Latent inhibition (indicated by a weaker preference in pre-exposed subjects) was observed in the experiment using the vanilla flavor. However, facilitation (a stronger preference in pre-exposed subjects) instead of latent inhibition was evident with the almond flavor, both across acquisition trials and in the final choice test. These results indicate that, unlike most other paradigms of Pavlovian conditioning, conditioned stimulus pre-exposure in flavor preference learning may either facilitate or retard the acquisition (or the expression) of a conditioned flavor preference. We explore the proposal that the critical difference between the flavors lies in their hedonic values, with facilitation being more likely in a flavor that is initially disliked.

Murine genetic variance in muscarinic cholinergic receptor antagonism of acquisition and expression of sucrose-conditioned flavor preferences in three inbred mouse strains

Conditioned flavor preferences (CFP) are elicited by sucrose relative to saccharin in inbred mice with both the robustness of the preferences and sensitivity to pharmacological receptor antagonists sensitive to genetic variance. Dopamine, opioid and N-methyl-d-aspartate receptor antagonists differentially interfere with the acquisition (learning) and expression (maintenance) of sucrose-CFP in BALB/c and SWR inbred mice. Further, the muscarinic cholinergic receptor antagonist, scopolamine (SCOP) more potently reduces both sucrose and saccharin intake in BALB/c and C57BL/6 relative to SWR inbred mice. The present study examined whether SCOP altered the expression and acquisition of sucrose-CFP in BALB/c, C57BL/6 and SWR mice. In expression experiments, food-restricted mice alternately consumed a flavored (CS+, e.g., cherry, 5 sessions) 16% sucrose solution and a differently-flavored (CS−, e.g., grape, 5 sessions) 0.05% saccharin solution. Two-bottle CS choice tests with the two flavors mixed in 0.2% saccharin solutions occurred following vehicle or SCOP at doses of 1 or 5 mg/kg. SCOP significantly reduced the magnitude of the expression of sucrose-CFP in BALB/c, but not either C57BL/6 or SWR mice. In acquisition experiments, separate groups of BALB/c, C57BL/6 and SWR mice were treated prior to acquisition training sessions with vehicle or 2.5 or 5 mg/kg SCOP doses that was followed by six two-bottle CS choice tests without injections. SCOP dose-dependently reduced (1 mg/kg) and eliminated (2.5 mg/kg) the acquisition of sucrose-CFP in BALB/c mice, and reduced the magnitude of acquisition of sucrose-CFP in SWR mice. In contrast, neither SCOP dose affected the acquisition of sucrose-CFP in C57BL/6 mice. Thus, muscarinic cholinergic receptor signaling is essential for the learning of sucrose-CFP in BALB/c mice, to a lesser degree in SWR mice, but not in C57BL/6 mice. Murine genetic variance differentially modulates muscarinic cholinergic receptor control of sweet intake per se relative to learned conditioned flavor preferences of sweets.

Orexin A in the ventral tegmental area enhances saccharin-induced conditioned flavor preference: The role of D1 receptors in central nucleus of amygdala

In industrialized societies, food intake is largely determined by its hedonic characteristics, which can be modified by our experience via taste learning. In this learning, the hedonic value of a neutral flavor changes after its association with a motivationally significant stimulus. Experiment 1 analyzes the effect of orexin administration (53 and 107 ng) in the ventral tegmental area (VTA) on hedonic intake through acquisition of a flavor-taste preference and a flavor-taste aversion. Accordingly, animals underwent four one-bottle acquisition sessions with unilateral application of orexin-A or saline in the VTA at 10 min before a 15-min flavor intake period. Preference and aversion were tested by a two-bottle test containing the flavors used for CS+ and CS−. Results indicate that intra-VTA orexin strengthens flavor-taste conditioned flavor preference (CFP) by saccharin but does not facilitate flavor-taste aversion induced by association of a neutral flavor with the unpalatable taste of quinine. Experiment 2 examines the acquisition of a flavor-taste preference after co-administration of an effective dose of orexin-A in the VTA and of D1-like dopamine receptor antagonist SCH23390 (6 and 12 nmol) in the central nucleus of the amygdala (CeA). SCH23390 impedes the CFP strengthening observed after intra-VTA orexin administration, indicating that this effect may be mediated by dopaminergic receptors in the CeA. These data suggest that the simultaneous presentation of a flavor and a hedonically positive taste may be detected by orexinergic neurons that activate dopamine-releasing neurons of the VTA, thereby reinforcing the positive signals required to develop a taste preference.

Prior exposure to nutritive and artificial sweeteners differentially alters the magnitude and persistence of sucrose-conditioned flavor preferences in BALB/c and C57BL/6 inbred mouse strains

Murine genetic variance affects sucrose’s ability to condition flavor preferences (CFP) relative to saccharin. Whereas BALB/c mice display robust sucrose- and fructose-CFP, C57BL/6 mice only display sucrose-CFP. Prior exposure to sucrose or saccharin solutions alters subsequent food choice responsiveness. The present study examined whether pre-exposure for one month to 10% sucrose or 0.2% saccharin altered subsequent sucrose-CFP in male and female BALB/c and C57BL/6 mice. Two weeks later, food-restricted mice were exposed to 10 CFP training trials with uniquely flavored 16% sucrose and 0.2% saccharin solutions. Two-bottle choice tests of the flavors mixed in saccharin followed for 4 weeks. Male mice weighed more than females across all conditions, and male BALB/c, but not C57BL mice consumed more 85 sucrose than females. No other notable sex differences were observed. BALB/c mice consumed more sucrose during pre-exposure and one-bottle training than C57BL/6 mice. Although the magnitudes of sucrose-CFP were comparable in two-bottle choice tests in water-exposed BALB/c and C57BL/6 mice, sucrose- and saccharin-exposed BALB/c mice displayed significantly greater sucrose-CFP preferences relative to C57BL/6 counterparts. These data indicate murine genetic variance in the effects of prior exposure to nutritive or non-nutritive sweeteners upon the magnitude of adult sugar-CFP.

Flavor preferences conditioned by nutritive and non-nutritive sweeteners in mice

Recent studies suggest that preferences are conditioned by nutritive (sucrose) but not by non-nutritive (sucralose) sweeteners in mice. Here we compared the effectiveness of nutritive and non-nutritive sweeteners to condition flavor preferences in three mouse strains. Isopreferred sucrose and sucralose solutions both conditioned flavor preferences in C57BL/6J (B6) mice but sucrose was more effective, consistent with its post-oral appetition action. Subsequent experiments compared flavor conditioning by fructose, which has no post-oral appetition effect in B6 mice, and a sucralose+saccharin mixture (SS) which is highly preferred to fructose in 24-h choice tests. Both sweeteners conditioned flavor preferences but fructose induced stronger preferences than SS. Training B6 mice to drink a flavored SS solution paired with intragastric fructose infusions did not enhance the SS-conditioned preference. Thus, the post-oral nutritive actions of fructose do not explain the sugar's stronger preference conditioning effect. Training B6 mice to drink a flavored fructose solution containing SS did not reduce the sugar-conditioned preference, indicating that SS does not have an off-taste that attenuates conditioning. Although B6 mice strongly preferred flavored SS to flavored fructose in a direct choice test, they preferred the fructose-paired flavor to the SS-paired flavor when these were presented in water. Fructose conditioned a stronger flavor preference than an isopreferred saccharin solution, indicating that sucralose is not responsible for the limited SS conditioning actions. SS is highly preferred by FVB/NJ and CAST/EiJ inbred mice, yet conditioned only weak flavor preferences. It is unclear why highly or equally preferred non-nutritive sweeteners condition weaker preferences than fructose, when all stimulate the same T1r2/T1r3 sweet receptor. Recent findings support the existence of non-T1r2/T1r3 glucose taste sensors; however, there is no evidence for receptors that respond to fructose but not to non-nutritive sweeteners.

Acquisition and expression of fat-conditioned flavor preferences are differentially affected by NMDA receptor antagonism in BALB/c and SWR mice

Conditioned flavor preferences are elicited by fat (Intralipid) in inbred mouse strains with BALB/c and SWR mice displaying among the most robust preferences. Dopamine D1 and opioid receptor antagonism differentially reduces the acquisition (learning) and expression (maintenance) of fat-conditioned flavor preferences in these two strains. Because noncompetitive NMDA receptor antagonism with MK-801 differentially altered sugar-conditioned flavor preferences in these strains, and because NMDA receptors are involved in fat intake, the present study examined whether MK-801 differentially altered expression and acquisition of fat (Intralipid)-conditioned flavor preferences in BALB/c and SWR mice. In expression studies, food-restricted male mice alternately consumed a flavored (CS+, e.g., cherry, 5 sessions) 5% Intralipid solution and a differently-flavored (CS−, e.g., grape, 5 sessions) 0.5% Intralipid solution. Two-bottle CS choice tests occurred following vehicle or MK-801 (100, 200µg/kg). MK-801 blocked expression of Intralipid-CFP at both doses in BALB/c mice, but only at the 100µg/kg dose in SWR mice. In acquisition studies, groups of BALB/c (0, 100µg/kg) and SWR (0, 100µg/kg) male mice were treated prior to the ten acquisition training sessions followed by six 2-bottle CS choice tests without injections. MK-801 eliminated acquisition of Intralipid-conditioned flavor preferences in BALB/c mice, and actually changed the preference to an avoidance response in SWR mice. Thus, NMDA receptor signaling appears essential especially for the learning of fat-conditioned flavor preferences in both mouse strains.