Olfaction contributes to the learned avidity for glucose relative to fructose in mice.

When offered glucose and fructose solutions, rodents consume more glucose solution because it produces stronger postoral reinforcement. Intake of these sugars also conditions a higher avidity for glucose relative to fructose. We asked which chemosensory cue mediates the learned avidity for glucose. We subjected mice to 18 days of sugar training, offering them 0.3, 0.6, and 1 M glucose and fructose solutions. Before and after training, we measured avidity for 0.3 and 0.6 M glucose and fructose in brief-access lick tests. First, we replicated prior work in C57BL/6 mice. Before training, the mice licked at a slightly higher rate for 0.6 M fructose; after training, they licked at a higher rate for 0.6 M glucose. Second, we assessed the necessity of the glucose-specific ATP-sensitive K+ (KATP) taste pathway for the learned avidity for glucose, using mice with a nonfunctional KATP channel [regulatory sulfonylurea receptor (SUR1) knockout (KO) mice]. Before training, SUR1 KO and wild-type mice licked at similar rates for 0.6 M glucose and fructose; after training, both strains licked at a higher rate for 0.6 M glucose, indicating that the KATP pathway is not necessary for the learned discrimination. Third, we investigated the necessity of olfaction by comparing sham-treated and anosmic mice. The mice were made anosmic by olfactory bulbectomy or ZnSO4 treatment. Before training, sham-treated and anosmic mice licked at similar rates for 0.6 M glucose and fructose; after training, sham-treated mice licked at a higher rate for 0.6 M glucose, whereas anosmic mice licked at similar rates for both sugars. This demonstrates that olfaction contributes significantly to the learned avidity for glucose.

Encoding of contextual fear memory requires de novo proteins in the prelimbic cortex.

BACKGROUND: Despite our understanding of the significance of the prefrontal cortex in the consolidation of long-term memories (LTM), its role in the encoding of LTM remains elusive. Here we investigated the role of new protein synthesis in the mouse medial prefrontal cortex (mPFC) in encoding contextual fear memory. METHODS: Because a change in the association of mRNAs to polyribosomes is an indicator of new protein synthesis, we assessed the changes in polyribosome-associated mRNAs in the mPFC following contextual fear conditioning (CFC) in the mouse. Differential gene expression in mPFC was identified by polyribosome profiling (n = 18). The role of new protein synthesis in mPFC was determined by focal inhibition of protein synthesis (n = 131) and by intra-prelimbic cortex manipulation (n = 56) of Homer 3, a candidate identified from polyribosome profiling. RESULTS: We identified several mRNAs that are differentially and temporally recruited to polyribosomes in the mPFC following CFC. Inhibition of protein synthesis in the prelimbic (PL), but not in the anterior cingulate cortex (ACC) region of the mPFC immediately after CFC disrupted encoding of contextual fear memory. Intriguingly, inhibition of new protein synthesis in the PL 6 hours after CFC did not impair encoding. Furthermore, expression of Homer 3, an mRNA enriched in polyribosomes following CFC, in the PL constrained encoding of contextual fear memory. CONCLUSIONS: Our studies identify several molecular substrates of new protein synthesis in the mPFC and establish that encoding of contextual fear memories require new protein synthesis in PL subregion of mPFC.

Ghrelin signaling is not essential for sugar or fat conditioned flavor preferences in mice.

The oral and post-oral actions of sugar and fat stimulate intake and condition flavor preferences in rodents through a process referred to as appetition. Ghrelin is implicated in food reward processing, and this study investigated its involvement in nutrient conditioning in mice. In Exp. 1 ghrelin receptor-null (GHSR-null) and C57BL/6 wildtype (WT) mice learned to prefer a flavor (CS+) mixed into 8% glucose over another flavor (CS-) mixed into a "sweeter" but non-nutritive 0.1% sucralose+saccharin (S+S) solution. In Exp. 2 treating WT mice with a ghrelin receptor antagonist [(D-Lys3)-GHRP-6] during flavor training did not prevent them from learning to prefer the CS+ glucose over the CS-S+S flavor. GHSR-null and WT mice were trained in Exp. 3 to drink a CS+ paired with intragastric (IG) infusion of 16% glucose and a CS- paired with IG water. Both groups drank more CS+ than CS- in training and preferred the CS+ to CS- in a choice test. The same (Exp. 4) and new (Exp. 5) GHSR-null and WT mice learned to prefer a CS+ flavor paired with IG fat (Intralipid) over a CS- flavor paired with IG water. GHSR-null and WT mice also learned to prefer a CS+ flavor added to 8% fructose over a CS- added to water. Together, these results indicate that ghrelin receptor signaling is not required for flavor preferences conditioned by the oral or post-oral actions of sugar and fat. This contrasts with other findings implicating ghrelin signaling in food reward processing and food-conditioned place preferences.

Intragastric fat self-administration is impaired in GPR40/120 double knockout mice.

Mice acquire strong preferences for flavors paired with intragastric (IG) fat infusions. This IG fat conditioning is attenuated in double knockout (DoKO) mice missing GPR40 and GPR120 fatty acid receptors. Here we determined if GPR40/120 DoKO mice are also impaired in IG fat self-administration in an operant lick task. In daily 1-h sessions the mice were trained with a sipper spout that contained dry food pellets; licks on the spout triggered infusions of IG fat (Intralipid). The training sessions were followed by test sessions with an empty spout. GPR40/120 DoKO mice self-infused more 20% fat than wild type (WT) C57BL/6 mice in training with a food-baited spout (2.4 vs. 2.0kcal/h) but self-infused less 20% fat than WT mice in empty spout tests (1.2 vs. 1.7kcal/h). The DoKO mice also self-infused less 5% fat than WT mice (0.6 vs. 1.3kcal/h) although both groups emitted more licks for 5% fat than 20% fat. The DoKO and WT mice did not differ, however, in their self-infusion of 12.5% glucose (1.5 vs. 1.6kcal/h), which is isocaloric to 5% fat. A second 5% IL test showed that the DoKO mice reverted to a reduced self-infusion compared to WT mice. When the infusion was shifted to water, WT mice reduced licking in the first extinction session, whereas DoKO mice were less sensitive to the absence of infused fat. Our results indicate that post-oral GPR40/120 signaling is not required to process IG fat infusions in food-baited spout training sessions but contributes to post-oral fat reinforcement in empty spout tests and flavor conditioning tests.

Role of NMDA, opioid and dopamine D1 and D2 receptor signaling in the acquisition of a quinine-conditioned flavor avoidance in rats.

A conditioned flavor preference (CFP) can be produced by pairing a flavor (conditioned stimulus, CS+) with the sweet taste of fructose. Systemic dopamine (DA) D1, D2 and NMDA, but not opioid, receptor antagonists significantly reduce the acquisition of the fructose-CFP. A conditioned flavor avoidance (CFA) can be produced by pairing a CS+flavor with the bitter taste of quinine. To evaluate whether fructose-CFP and quinine-CFA share common neurochemical substrates, the present study determined the systemic effects of DA D1 (SCH23390: SCH), DA D2 (raclopride: RAC), NMDA (MK-801) or opioid (naltrexone: NTX) receptor antagonists on the acquisition of quinine-CFA. In Experiment 1, food-restricted male rats were trained over 8 alternating one-bottle sessions to drink an 8% fructose+0.2% saccharin solution (FS) mixed with one flavor (CS-, e.g., grape) and a different flavor (CS+, e.g., cherry) mixed in a solution (FSQ) containing fructose+saccharin and quinine at 0.001-0.030% concentrations. In six subsequent two-bottle choice tests (1-3: two sessions each) with the CS- and CS+ flavors presented in FS solutions, only rats trained with 0.03% quinine displayed a CS+ avoidance in Test 1. In Experiment 2, rats received vehicle (Veh), SCH (200 nmol/kg), RAC (200 nmol/kg), MK-801 (100 µg/kg) or NTX (1 mg/kg) 30 min prior to the 8 one-bottle training sessions with CS-/FS and CS+/FSQ (0.03% quinine) solutions. An additional vehicle group (Veh 0.06%) was trained with a CS+/FSQ containing 0.06% quinine. In the two-bottle choice tests, the Veh and RAC groups avoided the CS+ flavor in Test 1 only, whereas the SCH, MK801, and NTX groups significantly avoided the CS+ in Tests 1-3. The Veh.06% group trained avoided the CS+ in Tests 1 and 2, but not Test 3. In Experiment 3, Veh and SCH groups were trained as in Experiment 2, but were tested with CS flavors presented in 0.2% saccharin solutions. The SCH group avoided the CS+ flavor in Tests 1-3 while the Veh group avoided the CS+ in Test 1 only. Thus whereas DA D1, DA D2 and NMDA, but not opioid receptor antagonism blocked acquisition of sweet taste-based CFP, DA D1, NMDA and opioid, but not DA D2 receptor antagonism enhanced the CFA produced by the bitter taste of quinine.

Effect of dopamine D1 and D2 receptor antagonism in the lateral hypothalamus on the expression and acquisition of fructose-conditioned flavor preference in rats.

The attraction to sugar-rich foods is influenced by conditioned flavor preferences (CFP) produced by the sweet taste of sugar (flavor-flavor learning) and the sugar's post-oral actions (flavor-nutrient) learning. Brain dopamine (DA) circuits are involved in both types of flavor learning, but to different degrees. This study investigated the role of DA receptors in the lateral hypothalamus (LH) on the flavor-flavor learning produced the sweet taste of fructose. In an acquisition study, food-restricted rats received bilateral LH injections of a DA D1 receptor antagonist (SCH23390), a D2 antagonist (RAC, raclopride) or vehicle prior to 1-bottle training sessions with a flavored 8% fructose+0.2% saccharin solution (CS+/F) and a less-preferred flavored 0.2% saccharin solution (CS-). Drug-free 2-bottle tests were then conducted with the CS+ and CS- flavors presented in saccharin. The fructose-CFP did not differ among groups given vehicle (76%), 12 nmol SCH (78%), 24 nmol (82%) or 24 nmol RAC (90%) during training. In an expression study with rats trained drug-free, LH injections of 12 or 24 nmol SCH or 12-48 nmol RAC prior to 2-bottle tests did not alter CS+ preferences (77-90%) relative to vehicle injection (86%). Only a 48 nmol SCH dose suppressed the CS+ preference (61%). The minimal effect of LH DA receptor antagonism upon fructose flavor-flavor conditioning differs with the ability of LH SCH injections to block the acquisition of glucose flavor-nutrient learning.

Dopamine D1 and opioid receptor antagonism effects on the acquisition and expression of fat-conditioned flavor preferences in BALB/c and SWR mice.

Sugar and fat appetites are influenced by unlearned and learned responses to orosensory and post-ingestive properties which are mediated by dopamine (DA) and opioid transmitter systems. In BALB and SWR mice, acquisition and expression of sucrose conditioned flavor preferences (CFP) are differentially affected by systemic DA D1 (SCH23390: SCH) and opioid (naltrexone: NTX) antagonism. The present study examined whether fat-CFP occurred in these strains using preferred (5%) and less preferred (0.5%) Intralipid solutions, and how SCH and NTX altered its expression and acquisition. Food-restricted mice drank flavored (CS+, e.g., cherry) 5% and flavored (CS-, e.g., grape) 0.5% Intralipid solutions in alternate 1 h/day sessions. Six two-bottle tests with both CS flavors presented in 0.5% Intralipid occurred. Robust and comparable fat-CFP was observed in both strains. In Experiment 2, vehicle (Veh), SCH (50-800 nmol/kg) or NTX (1-5 mg/kg) were administered prior to two-bottle testing in an identical paradigm. Expression of fat-CFP was significantly reduced by SCH and NTX in BALB mice, and only by SCH in SWR mice. NTX produced significantly greater inhibition of fat-CFP expression in BALB versus SWR mice. In Experiment 3, separate groups of BALB and SWR mice received Veh, SCH (50 nmol/kg) or NTX (1 mg/kg) injections prior to daily one-bottle CS+ and CS- training sessions to assess acquisition effects. Six two-bottle CS+ vs. CS- choice tests were then conducted. Fat-CFP acquisition was significantly reduced in SWR mice by SCH, but not NTX, and was only mildly affected by both antagonists in BALB mice. SCH produced significantly greater inhibition of fat-CFP acquisition in SWR versus BALB mice. The pattern of SCH effects upon the expression and acquisition of fat-CFP in BALB and SWR mice was quite similar to that observed for sucrose-CFP, suggesting similar DA D1 receptor involvement in sugar- and fat-CFP in these mouse strains.

Glucose-conditioned flavor preference learning requires co-activation of NMDA and dopamine D1-like receptors within the amygdala.

The role of amygdala (AMY) NMDA receptor signaling and its interaction with dopamine D1-like receptor signaling in glucose-mediated flavor preference learning was investigated. In Experiment 1, rats were trained with a flavor (CS+) paired with intragastric (IG) 8% glucose infusions and a different flavor (CS-) paired with IG water infusions. In the two-bottle tests (Expression), bilateral intra-AMY injections of the NMDA receptor antagonist, AP5 (0, 5 and 10 nmol/brain), did not block the CS+ preference. In Experiment 2, new rats received intra-AMY injections of either vehicle or AP5 (10 nmol), prior to training sessions with CS+/IG glucose and CS-/IG water. In the two-bottle tests without drug treatment, AP5 rats failed to prefer the CS+ flavor (50%). In Experiments 3, new rats were trained as in Experiment 2 except that, during training, half the rats received AP5 injections (5 nmol) in one side of the AMY and SCH23390 (D1-like receptor antagonist, 6 nmol), in the contralateral AMY (Drug/Drug group). The remaining rats received vehicle injections in one side of the AMY and either AP5 (5 nmol) or SCH23390 (6 nmol) in the contralateral AMY (Drug/Vehicle group). The two-bottle choice tests without drug treatment revealed that, unlike the Drug/Vehicle group (85%), the Drug/Drug group failed to prefer the CS+ flavor (50%). These results reveal an essential role for AMY NMDA receptor activation in the acquisition of flavor preference learning induced by the post-oral reinforcing properties of glucose and demonstrate that such learning is based on co-activation of NMDA and DA D1 receptors within this forebrain structure.

Double-dissociation of D1 and opioid receptor antagonism effects on the acquisition of sucrose-conditioned flavor preferences in BALB/c and SWR mice.

Sugar appetite is influenced by unlearned attractions to sweet taste and learned responses to sugars' taste and post-ingestive actions. In rats, sugar-conditioned flavor preferences (CFP) are attenuated by dopamine D1 (SCH23390: SCH), but not by opioid (naltrexone: NTX), receptor antagonism. Sucrose-CFP occurs in BALB/c and SWR inbred mice that differ in their suppressive effects of SCH and NTX on sucrose intake. The present study examined whether SCH and NTX altered expression of previously learned sucrose-CFP and acquisition (learning) of sucrose-CFP in these strains. In Experiment 1, food-restricted mice were trained (10 one-bottle sessions) to drink a more-preferred flavored (e.g., cherry) 16% sucrose solution (CS+/Sucrose) on odd-numbered days, and a less-preferred flavored (e.g., grape) 0.05% saccharin solution (CS-/Saccharin) on even-numbered days. Two-bottle tests with the flavors mixed in 0.2% saccharin occurred 30 min following vehicle (Veh), SCH (50-800 nmol/kg) or NTX (1-5mg/kg) assessing preference expression. CS+ preference expression in BALB/c and SWR mice following Veh were significantly reduced by SCH and NTX. In Experiment 2, separate groups of BALB/c and SWR mice received Veh, SCH (50 nmol/kg) or NTX (1mg/kg) injections 30 min prior to daily one-bottle training sessions with the CS+/Sucrose and CS-/Saccharin solutions assessing preference acquisition. Subsequent two-bottle tests with the CS+ vs. CS- solutions were conducted without injections. CS+/Sucrose training intakes were reduced by SCH in both strains and by NTX in BALB/c mice. In the initial two-bottle test, sucrose-CFP acquisition was significantly reduced in BALB NTX (54%), but not in BALB SCH (77%) groups relative to the BALB Veh group (85%). In contrast, sucrose-CFP acquisition was significantly reduced in SWR SCH (61%), but not in SWR NTX (83%) groups relative to the SWR Veh group (86%). DA D1 and opioid receptor signaling modulate acquisition and/or expression of sucrose-CFP in mice with significant strain differences observed.

Dopamine signaling in the medial prefrontal cortex and amygdala is required for the acquisition of fructose-conditioned flavor preferences in rats.

Systemic administration of dopamine (DA) D1 (SCH23390: SCH) and D2 (raclopride: RAC) antagonists blocked both acquisition and expression of fructose-conditioned flavor preferences (CFP). It is unclear what brain circuits are involved in mediating these effects. The present study investigated DA signaling within the nucleus accumbens shell (NAcS), amygdala (AMY) and medial prefrontal cortex (mPFC) in the acquisition and expression of fructose-CFP. In Experiment 1, separate groups of rats were injected daily in the NAcS or AMY with saline, SCH (24 nmol) or RAC (24 nmol) prior to training sessions with a flavor (CS+) mixed with 8% fructose and 0.2% saccharin (CS+/F) and a different flavor (CS-) mixed with only 0.2% saccharin. In the two-bottle choice tests with 0.2% saccharin, only rats injected with RAC in the AMY failed to acquire a CS+ preference (45-54%). In Experiment 2, new rats were identically trained, but saline, SCH and RAC were injected in the mPFC. In subsequent two-bottle choice tests, SCH- and RAC-treated rats failed to exhibit a CS+ preference (50-56%). In Experiment 3, new rats were trained with CS+/F and CS- without injections. Subsequent two-bottle choice tests were then conducted following bilateral injections of SCH or RAC in the mPFC at total doses of 0, 12, 24 and 48 nmol. Expression of the CS+ preference failed to be affected by either antagonist, indicating that the mPFC is not involved in the maintenance of this preference. These data indicate that the acquisition of fructose-CFP is dependent on DA signaling in the mPFC and AMY.

Strain differences in sucrose- and fructose-conditioned flavor preferences in mice.

Genetic factors strongly influence the intake and preference for sugar and saccharin solutions in inbred mouse strains. The present study determined if genetic variance also influences the learned preferences for flavors added to sugar solutions. Conditioned flavor preferences (CFPs) are produced in rodents by adding a flavor (CS+) to a sugar solution and a different flavor (CS-) to a saccharin solution (CS-) in one-bottle training trials; the CS+ is subsequently preferred to the CS- when both are presented in saccharin solutions in two-bottle tests. With some sugars (e.g., sucrose), flavor preferences are reinforced by both sweet taste and post-oral nutrient effects, whereas with other sugars (e.g., fructose), sweet taste is the primary reinforcer. Sucrose and fructose were used in three experiments to condition flavor preferences in one outbred (CD-1) and eight inbred strains which have "sensitive" (SWR/J, SJL/J, C57BL/10J, C57BL/6J) or "sub-sensitive" (DBA/2J, BALB/cJ, C3H/HeJ, 129P3/J) sweet taste receptors (T1R2/T1R3). Food-restricted mice of each strain were trained (1 h/day) to drink flavored 16% sucrose (CS+ 16S, Experiment 1), 16% fructose (CS+ 16F, Experiment 2) or 8% fructose+0.2% saccharin (CS+ 8F, Experiment 3) solutions on five alternate days and a differently flavored saccharin solution (0.05% or 0.2%, CS-) on the other five alternating days. The CS+ and CS- flavors were presented in 0.2% saccharin for two-bottle testing over six days. All strains preferred the CS+ 16S to CS- although there were significant strain differences in the magnitude and persistence of the sucrose preference. The strains also differed in the magnitude and persistence of preferences for the CS+ 16F and CS+ 8F flavors over the CS- with two strains failing to prefer the fructose-paired flavors. Sucrose conditioned stronger preferences than did fructose which is attributed to differences in the taste and post-oral actions of the sugars. These differential training intakes may not have influenced the sucrose-CFP because of the post-oral reinforcing actions of sucrose. Overall, sweet sensitive and sub-sensitive mice did not differ in sucrose-CFP, but unexpectedly, the sub-sensitive mice displayed stronger fructose-CFP. This may be related to differential training intakes of CS+ and CS- solutions: sweet sensitive mice consumed more CS- than CS+ during training while sub-sensitive mice consumed more CS+.

Dopamine and learned food preferences.

An early study performed in Bart Hoebel's laboratory suggested that dopamine (DA) signaling in the nucleus accumbens was involved in learned flavor preferences produced by post-oral nutritive feedback. This paper summarizes our studies investigating the role of DA in flavor preference conditioning using selective DA receptor antagonists. Food-restricted rats were trained to prefer a flavored saccharin solution (CS+) paired with intragastric (IG) sugar infusions over a flavored saccharin solution (CS-) paired with water infusions. Systemic injections of a D1-like receptor antagonist (SCH23390), but not a D2-like receptor antagonist (raclopride) during training blocked flavor preference learning. Neither drug prevented the expression of an already learned preference except at high doses that greatly suppressed total intakes. Central sites of action were examined by local microinjections of SCH23390 (12 nmol) during flavor training or testing. Drug infusions in the nucleus accumbens, amygdala, medial prefrontal cortex, or lateral hypothalamus during training blocked or attenuated CS+ flavor conditioning by IG glucose infusions. The same drug dose did not suppress the expression of a learned CS+ preference. The findings suggest that DA signaling within different components of a distributed brain network is involved in sugar-based flavor preferences. A possible role of DA in conditioned increases in flavor acceptance is discussed.

Opioid mediation of starch and sugar preference in the rat.

In our prior studies, administration of the opioid receptor antagonist naltrexone did not block conditioned preferences for a flavor paired with a preferred sugar solution over a flavor paired with saccharin. This may be because both training solutions were sweet, and their attractiveness was reduced by naltrexone. The present study compared the effects of naltrexone on preferences for flavors paired with sugar or starch drinks that have distinctive tastes to rats. Experiment 1 assessed naltrexone's effect on the preference for unflavored 8% cornstarch and 8% sucrose aqueous solutions/suspensions. The food-restricted rats displayed a significant sucrose preference which increased following systemic treatment with naltrexone (1 or 3mg/kg) even though total intake of both solutions declined. In Experiment 2, rats were trained to drink flavored (cherry or grape) starch and sucrose solutions in separate one-bottle sessions. In a two-bottle choice test with both flavors presented in a sucrose-starch mixture, the rats significantly preferred the starch-paired flavor. Naltrexone treatment blocked the expression of this starch-conditioned preference. In Experiment 3, rats were treated with saline or naltrexone throughout one-bottle training with flavored sucrose and starch solutions. In a subsequent choice test, both the saline and naltrexone groups displayed significant preferences for the starch-paired flavor, indicating that opioid antagonism failed to alter the acquisition of this conditioned preference. In summary, novel outcomes of this study included the increased rather than the predicted decrease in sucrose preference produced by naltrexone. Also, starch unexpectedly conditioned the stronger flavor preference, although this can be explained by the differential post-oral reinforcing actions of starch and sucrose, and naltrexone blocked the expression, but not the acquisition, of this preference. These findings suggest that the reward value of starch in liquid form is more dependent upon opioid signaling than is that of sugar.

Neuropharmacology of learned flavor preferences.

Innate and learned flavor preferences influence food and fluid choices in animals. Two primary forms of learned preferences involve flavor-flavor and flavor-nutrient associations in which a particular flavor element (e.g., odor) is paired with an innately preferred flavor element (e.g., sweet taste) or with a positive post-oral nutrient consequence. This review summarizes recent findings related to the neurochemical basis of learned flavor preferences. Systemic and central injections of dopamine receptor antagonists implicate brain dopamine signaling in both flavor-flavor and flavor-nutrient conditioning by the taste and post-oral effects of sugars. Dopamine signaling in the nucleus accumbens, amygdala and lateral hypothalamus is involved in one or both forms of conditioning and selective effects are produced by D1-like and D2-like receptor antagonism. Opioid receptor antagonism, despite its suppressive action on sugar intake and reward, has little effect on the acquisition or expression of flavor preferences conditioned by the sweet taste or post-oral actions of sugars. Other studies indicate that flavor preference conditioning by sugars is differentially influenced by glutamate receptor antagonism, cannabinoid receptor antagonism and benzodiazepine receptor activation.

Acquisition of glucose-conditioned flavor preference requires the activation of dopamine D1-like receptors within the medial prefrontal cortex in rats.

In this study, we investigated the role of dopamine transmission within the medial prefrontal cortex (mPFC) in flavor preference learning induced by post-oral glucose. In Experiment 1, rats were trained with a flavor (CS+) paired with intragastric (IG) infusions of 8% glucose and a different flavor (CS-) paired with IG water infusions. The CS+ preference was evaluated in two-bottle tests following bilateral injection of the dopamine D1-like receptor antagonist, SCH23390, into the mPFC at total doses of 0, 12 and 24nmol. SCH23390 produced dose-dependent reductions in CS+ intake but did not block the CS+ preference. In Experiment 2, new rats were injected daily in the mPFC with either saline or SCH23390 (12nmol), prior to training sessions with CS+/IG glucose and CS-/IG water. In the two-bottle choice tests, SCH rats, unlike the Control rats, failed to prefer the CS+ (50% vs. 74%). Collectively, the results show that D1-like receptor activation in the medial prefrontal cortex plays a crucial role in the acquisition of flavor preference learning induced by the post-oral reinforcing properties of glucose.

Opioid receptor antagonism in the nucleus accumbens fails to block the expression of sugar-conditioned flavor preferences in rats.

In our prior studies, systemic administration of the opioid receptor antagonist naltrexone (NTX) did not block flavor preference conditioning by the sweet taste or post-oral actions of sugar despite reducing intake. Because opioid signaling in the nucleus accumbens (NAc) is implicated in food reward, this study determined if NTX administered into the NAc would block the expression of sugar-conditioned preferences. In Experiment 1, food-restricted rats with bilateral NAc shell or core cannulae were trained to drink a fructose (8%)+saccharin (0.2%) solution mixed with one flavor (CS+) and a less-preferred 0.2% saccharin solution mixed with another flavor (CS-) during one-bottle sessions. Two-bottle tests with the two flavors mixed in saccharin solutions occurred 10 min following total bilateral NAc shell or core doses of 0, 1, 25 and 50 microg of NTX. The rats preferred the CS+ over CS- following vehicle (80%) and all NTX doses in the shell and core. The CS+ preference was reduced to 64% and 72% by 50 microg NTX in the shell and core, although only the core effect was significant. In Experiment 2, food-restricted rats were trained to drink one flavored saccharin solution (CS+) paired with an intragastic (IG) glucose (8%) infusion and a second flavored saccharin solution (CS-) paired with an IG water infusion. In subsequent two-bottle tests, the rats displayed significant preferences for the CS+ (81-91%) that were unaltered by any NTX dose in the shell or core. CS+ intake, however, was reduced by NTX in the shell, but not the core. These data indicate that accumbal opioid antagonism slightly attenuated, but did not block the expression of sugar-conditioned flavor preferences. Therefore, while opioid drugs can have potent effects on sugar intake they appear less effective in altering sugar-conditioned flavor preferences.

Role of olfaction in the conditioned sucrose preference of sweet-ageusic T1R3 knockout mice.

Prior work has shown that sweet taste-deficient T1R3 knockout (KO) mice developed significant sucrose preferences when given long-term sugar versus water tests. The current study investigated the role of olfaction in this experience-conditioned sucrose preference. T1R3 KO and C57BL/6 wild-type (WT) mice were given 24-h sugar versus water tests with ascending concentrations of sucrose (0.5-32%), after which the mice received olfactory bulbectomy (OBx) or sham surgery. When retested with sucrose, the Sham-KO mice preferred all sugar solutions to water, although their intake and preference were less than those of the Sham-WT mice. The OBx-KO mice, in contrast, showed no or weak preferences for dilute sucrose solutions (0.5-8%) although they strongly preferred concentrated sugar solutions (16-32%). OBx-WT mice displayed only a partial reduction in their sucrose preference. Although the OBx mice of both genotypes underconsumed dilute sucrose solutions relative to Sham mice, they overconsumed concentrated sucrose. These results indicate that olfaction plays a critical role in the conditioned preference of T1R3 KO mice for dilute sugar solutions. Further, the fact that OBx-KO mice preferred concentrated sucrose solutions in the absence of normal sweet taste and olfactory sensations underscores the potency of postoral nutritive signals in promoting ingestion.

Dopamine D1-like receptor antagonism in amygdala impairs the acquisition of glucose-conditioned flavor preference in rats.

This study examined the role of dopamine within the amygdala (AMY) in flavor preference learning induced by post-oral glucose. In Experiment 1, rats were trained with a flavor [conditioned stimulus (CS+)] paired with intragastric (IG) infusions of 8% glucose and a different flavor (CS-) paired with IG water infusions. The CS+ preference was evaluated in two-bottle tests following bilateral injection of the dopamine D1-like receptor antagonist, SCH23390 (SCH), into the AMY at total doses of 0, 12, 24 and 48 nmol. SCH produced dose-dependent reductions in CS+ intake but did not block the CS+ preference except at the two highest doses, which also greatly suppressed the CS intakes. In Experiment 2, new rats were injected daily in the AMY with either saline or SCH (12 nmol), prior to training sessions with CS+/IG glucose and CS-/IG water. In the two-bottle tests, SCH rats, unlike the control rats, failed to prefer the CS+ (55 vs. 81%). In Experiments 3 and 4, new rats were trained as in Experiment 2, except that brain injections were in the basolateral and central nuclei of the AMY, respectively. SCH rats learned to prefer the CS+ to the CS-, although their preference was weaker than that displayed by the control rats (Experiment 3: 59 vs. 80%; Experiment 4: 73 vs. 88%). These results show an essential role for D1-like receptor activation in the AMY in the acquisition of flavor preference learning induced by the post-oral reinforcing properties of glucose. A distributed network mediating flavor-nutrient incentive learning is discussed.

Role of amygdala dopamine D1 and D2 receptors in the acquisition and expression of fructose-conditioned flavor preferences in rats.

Systemic administration of dopamine D1-like (SCH23390) and, to a lesser degree D2-like (raclopride), receptor antagonists significantly reduce the acquisition and expression of fructose-conditioned flavor preferences (CFP) in rats. Given the role of dopamine in the amygdala (AMY) in the processing and learning of food reward, the present study examined whether dopamine D1-like or D2-like antagonists in this site altered acquisition and/or expression of a fructose-CFP. In Experiment 1, food-restricted rats with bilateral AMY cannulae were trained to drink a fructose (8%)+saccharin (0.2%) solution mixed with one flavor (e.g., grape, CS+/Fs) and a less-preferred 0.2% saccharin solution mixed with another flavor (e.g., cherry, CS-/s) during one-bottle (16 ml) sessions. Two-bottle tests with the two flavors mixed in saccharin solutions (CS+/s, CS-/s) occurred 10 min following total bilateral AMY doses of 0, 12, 24 and 48 nmol of SCH23390 or raclopride. Preference for CS+/s over CS-/s was significantly reduced relative to vehicle baseline by the 48 nmol doses of SCH23390 and raclopride (from 77% to 66% and 68%), but not lower doses. In Experiment 2, rats received bilateral AMY injections (12 nmol) of SCH23390 (D1 group) or raclopride (D2 group) 10 min prior to one-bottle training sessions with CS+/Fs and CS-/s. Yoked Control rats received vehicle and were limited to the CS intakes of the D1 and D2 groups; untreated controls were not injected or limited to drug group intakes during training. Subsequent two-bottle tests revealed initial preferences of CS+/s over CS-/s in all groups that remained stable in untreated and Yoked Controls, but were lost over the 6 tests sessions in the D1 group, but not in the D2 group. These data indicate that dopamine D1-like and D2-like antagonists significantly attenuated the expression of the previously acquired fructose-CFP, and did not block acquisition of the fructose-CFP. D1-like antagonism during training hastened extinction of the fructose-CFP. The results are similar to those produced by dopamine D1-like and D2-like antagonist injections into the nucleus accumbens shell which suggests that flavor conditioning involves a regionally distributed brain network.

Lateral hypothalamus dopamine D1-like receptors and glucose-conditioned flavor preferences in rats.

This study examined the role of dopamine D1-like receptor transmission in the lateral hypothalamus (LH) in flavor preference learning induced by intragastric (IG) infusions of glucose. Rats fitted with gastric catheters were injected daily in the LH with either saline or SCH23390 (12 nmol/brain), 10 min prior to training sessions with a flavor (CS+) paired with IG infusions of 8% glucose and a different flavor (CS-) paired with IG water infusions. In a post-training two-bottle test, SCH-treated rats preferred the CS+ to the CS- although their preference was weaker than that of the Control rats (61% vs. 87%). The same dose of SCH23390 reduced CS+ intake of the Control rats in a subsequent test but did not suppress their CS+ preference (90%). These results show that D1-like receptor activation in the lateral hypothalamus modulates the acquisition, but not the expression of flavor preference learning induced by the post-oral reinforcing properties of glucose.