Sucrose intake: Increase in non-stressed rats and reduction in chronically stressed rats are both prevented by the gamma-hydroxybutyrate (GHB) analogue, GET73

It has been previously shown that the gamma-hydroxybutyrate analogue N-(4-trifluoromethylbenzyl)-4-methoxybutanamide (GET73) inhibits consumption and reinforcing effect of palatable food, in rats, at doses that have no detrimental effect on open-field behaviour. Here we show that GET73 is also able to prevent both the development of preference for a sucrose solution in non-stressed rats, and the reduction of preference for a sucrose solution induced by the daily exposure to continuously varied mildly stressful situations. Adult male Wistar Kyoto rats (180-190 g) were subjected to chronic unpredictable mild stress. Other rats of the same sex and strain were used to study the development of preference for a sucrose solution. Daily exposure to continuously varied mildly stressful situations produced a reduction of sucrose solution intake that started the 3rd week, and such reduction became highly significant during the 5th week. Treatment with GET73 (10 mg kg(-1), 50 mg kg(-1) or 100 mg kg(-1) once daily per os) produced a more evident reduction of sucrose solution intake during the 2nd and 3rd week, but during the 4th and 5th weeks the intake dose-dependently increased to values that, for the dose of 100 mg kg(-1), were not significantly different from those of non-stressed, vehicle-treated rats. In the same range of doses GET73 dose-dependently prevented the development of preference for a sucrose solution in non-stressed rats. The present data indicate that rats treated with GET73 do not develop the "depression-like" condition produced by the daily exposure, for several weeks, to continuously and unpredictably varied stressful situations in a valid (face, predictive, and construct validity) "depression" model. Moreover, GET73 prevents the development of preference for a sucrose solution in non-stressed rats. Concurrently, present and previous data suggest that GET73 "stabilize" the behaviour of rats, either preventing the development of a "depression-like" condition in a continuously stressful environment, or the rewarding effect of alcohol, sucrose, and palatable food.     

Hindbrain orexin 1 receptors influence palatable food intake, operant responding for food, and food-conditioned place preference in rats

Rationale

Brain orexin 1 receptors (OX1Rs) are involved in food-motivated behavior. Most research has focused on forebrain OX1R populations, but hindbrain OX1Rs affect feeding. We hypothesized that hindbrain OX1Rs affect the reward value of food.

Objectives

We examined the effects of hindbrain OX1R stimulation or blockade on motivation for food, palatable high-fat (HF) food intake, and food-conditioned place preference.

Methods

Rats trained to lever press for sucrose on a progressive ratio (PR) schedule received fourth intracerebroventricular (icv) injections of vehicle, orexin-A (0.1–1 nmol), or the OX1R antagonist SB334867 (10–20 nmol) before operant test sessions. Effects of these treatments on HF food intake during daily 1-h tests were assessed with fourth icv and nucleus of the solitary tract (NTS) injections. We conditioned a place preference by pairing HF food with one side of a two-sided chamber and then examined the effect of 20 nmol fourth icv SB334867 on the expression of that preference.

Results

In ad lib fed rats on the PR schedule, fourth icv orexin-A significantly increased responding and breakpoint relative to the vehicle. In 24-h food-deprived rats, fourth icv SB334867 significantly decreased responding and breakpoint. Orexin-A delivered to the fourth ventricle (0.1 nmol) or NTS (0.01 nmol) increased HF diet intake. Fourth icv SB334867 did not affect HF food intake, but SB334867 delivered either fourth icv (20 nmol) or intra-NTS (5–10 nmol) suppressed chow intake. Expression of HF food-conditioned place preference was inhibited by fourth icv SB334867.

Conclusions

Hindbrain OX1R activity affects food-motivated operant behavior and may play a role in responding to cues that predict palatable food.     

Parathion-provoked lethality in rats is reduced by diethyldithiocarbamate

Intoxication of male rats with 5 mg/kg parathion reduces survival to 15%. Diethyldithiocarbamate (DDC), known to inhibit mixed-function oxidase activity, has no effect on survival rate when this compound is used in a dosage of 300 mg/kg 45 min before or 10 min after parathion intoxication. However, when DDC and parathion are administered simultaneously, the rate of survival rises to 53%.     

Intravenous cocaine self-administration in rats is reduced by dietaryl-tryptophan

Rats were trained to self-administer intravenously-delivered cocaine. Four lever-press responses resulted in a cocaine infusion (0.2 mg/kg) during daily 24-h sessions. The rats were also trained to obtain water from tongue-operated solenoid-driven drinking spouts. Ground food and water from a standard drinking bottle were also available. When cocaine injections reached stable levels,l-tryptophan was mixed with the rats' food for 5 days. Three concentrations ofl-tryptophan (2, 4, and 8%) were tested in different groups of five rats each. Three other groups of five rats each received the samel-tryptophan treatments; however, in these rats saline was substituted for cocaine and a sweet drinking solution consisting of glucose and saccharin (G + S) replaced water in the automatic drinking device. Two other groups consisting of five rats each self-administered a higher (0.4 mg/kg) or lower (0.1 mg/kg) unit dose of cocaine and food adulterated with 4% tryptophan. At the two higher concentrationsl-tryptophan reduced cocaine infusions by at least 50% during the 5 days of treatment, and cocaine infusions returned to baseline levels within 48 h after the regular diet was restored. Responding reinforced by the G + S solution was not altered by any of thel-tryptophan concentrations. Food intake was substantially lowered by the 8%l-tryptophan concentration; however, water intake, responding on an inactive lever, and the number of saline infusions were not affected by addition ofl-tryptophan to the food.l-Tryptophan had the same magnitude of effect on self-administration of the 0.1 and 0.2 mg/kg unit doses of cocaine, but behavior maintained by the highest cocaine dose (0.4 mg/kg) was resistant to the effect ofl-tryptophan. The results of this experiment indicate thatl-tryptophan reduces behavior reinforced by IV cocaine infusions.     

Withdrawal from chronic, intermittent access to a highly palatable food induces depressive-like behavior in compulsive eating rats

The increased availability of highly palatable foods is a major contributing factor toward the development of compulsive eating in obesity and eating disorders. It has been proposed that compulsive eating may develop as a form of self-medication to alleviate the negative emotional state associated with withdrawal from highly palatable foods. This study was aimed at determining whether withdrawal from chronic, intermittent access to a highly palatable food was responsible for the emergence of depressive-like behavior. For this purpose, a group of male Wistar rats was provided a regular chow diet 7 days a week (Chow/Chow), whereas a second group of rats was provided chow for 5 days a week, followed by a 2-day access to a highly palatable sucrose diet (Chow/Palatable). Following 7 weeks of diet alternation, depressive-like behavior was assessed during withdrawal from the highly palatable diet and following renewed access to it, using the forced swim test, the sucrose consumption test, and the intracranial self-stimulation threshold procedure. It was found that Chow/Palatable rats withdrawn from the highly palatable diet showed increased immobility time in the forced swim test and decreased sucrose intake in the sucrose consumption test compared with the control Chow/Chow rats. Interestingly, the increased immobility in the forced swim test was abolished by renewing access to the highly palatable diet. No changes were observed in the intracranial self-stimulation threshold procedure. These results validate the hypothesis that withdrawal from highly palatable food is responsible for the emergence of depressive-like behavior, and they also show that compulsive eating relieves the withdrawal-induced negative emotional state.     

Preference for cocaine by the weight pulling method in rats

The purpose of the present study is to show the efficiency of the weight pulling method in evaluating quantitatively the positive reinforcing effect of cocaine. Rats were trained to pull the weight in order to eat the drug-admixed food (DAF). The experiments began with the preexposure of the drug through the repetition of CFF schedule. The schedule consisted of one choice trial (C) between the intake of normal food and DAF followed by two consecutive forced trials (F), in which the rats were forced to take the DAF only. The study consisted of Experiment I, where cocaine concentration in DAF was varied while the period of cocaine preexposure was kept constant and Experiment II, where the period of preexposure was varied while the cocaine concentration was kept constant. Results show that the reinforcing effect of cocaine was dependent on cocaine intake. On the other hand, the reinforcing effect of cocaine was independent of cocaine preexposure period. The effect of cocaine on the drug-seeking behavior was evident on the first day of cocaine exposure. It is concluded that the weight pulling method is sufficient to evaluate quantitatively the reinforcing effects of cocaine in rats, and this method may be useful for the prediction of dependence potential in man.     

Preference for a cocaine-associated environment is attenuated by augmented accumbal serotonin in cocaine withdrawn rats

RATIONALE: Recent studies have found decreased serotonin (5-HT) transmission within the nucleus accumbens following withdrawal from chronic cocaine. OBJECTIVE: We sought to investigate whether increasing brain 5-HT levels would decrease behavioral responses that occur following cocaine withdrawal, namely increased preference for a cocaine environment and anxiety. METHODS: The conditioned place preference and the defensive burying paradigms were used to measure the behavioral responses that occur 1 week following cocaine withdrawal. RESULTS: We show that pharmacological agents that increase 5-HT transmission (sertraline or 5-hydoxytryptophan, 5-HTP) abolish the preference of subchronically cocaine-treated, abstinent rats for a cocaine-associated environment. Similar results were seen when sertraline was microinjected into the nucleus accumbens. Conversely, rats acutely conditioned with cocaine showed an increased preference for a cocaine-associated environment when pretreated with these drugs. Sertraline also decreased the heightened anxiety-like behaviors found in subchronically treated cocaine rats. CONCLUSIONS: These results indicate that drugs that augment 5-HT function may reduce the desire for cocaine following cocaine withdrawal, and thus facilitate cocaine abstinence in dependent subjects.     

Differential actions of dopamine receptor antagonism in rats upon food intake elicited by either mercaptoacetate or exposure to a palatable high-fat diet.

Selective dopamine receptor antagonists have been shown to reduce food intake of rats under such regulatory challenge conditions as food deprivation and 2-deoxy-D-glucose-induced glucoprivation, and under such palatable conditions as acute exposure to sucrose solutions. Food intake is increased following either pretreatment with the free fatty acid oxidation inhibitor, mercaptoacetate (MA), or acute exposure to a palatable high-fat source. The present study examined whether equimolar doses (50-800 nmol/kg, s.c.) of either the selective D(1) receptor antagonist, SCH23390, or the selective D(2) receptor antagonist, raclopride, would alter food intake elicited by either MA (70 mg/kg, i.p.) or acute exposure to a high-fat diet (67% ground rat chow, 33% vegetable shortening). SCH23390 significantly and dose-dependently reduced MA-induced feeding with the two higher (400 and 800 nmol/kg) doses eliminating this response after the first 2 h and the two lower (50 and 200 nmol/kg) doses preventing the occurrence of significant MA-induced feeding. Raclopride eliminated MA-induced feeding at the highest dose, and produced dose-dependent reductions at lower doses. A different pattern of dopamine antagonist effects emerged for high-fat intake. The identical dose range of SCH23390 failed to alter high-fat intake. In contrast, whereas the highest (800 nmol/kg) dose of raclopride significantly reduced high-fat intake after 1 h, the middle (200 and 400 nmol/kg) doses of raclopride significantly increased high-fat intake after 2 h. These data are discussed in terms of the modulatory actions of dopamine upon food intake, of the differential actions of dopamine receptor subtypes upon intake under challenge and palatable conditions, and of the potential participation of presynaptic and postsynaptic receptor populations in these responses.     

Olanzapine and sibutramine have opposing effects on the motivation for palatable food

Both olanzapine and sibutramine target serotonergic and noradrenergic neurotransmission and influence body weight, but in opposite ways. The second-generation antipsychotic olanzapine, an antagonist at serotonergic and noradrenergic receptors, frequently induces weight gain as a side-effect, whereas sibutramine, a noradrenaline/serotonin reuptake inhibitor, is known as a weight-reducing agent. To investigate whether altered motivation for palatable food influences the effect of these drugs on body weight, we determined their effects on responding for sucrose pellets under a progressive ratio schedule of reinforcement in rats. We found that a low dose of olanzapine selectively increased responding to sucrose, without affecting free-feeding intake of sucrose. In contrast, sibutramine dose-dependently reduced responding to sucrose and similarly reduced free-feeding intake. Furthermore, coadministration of a dose of sibutramine that failed to affect responding to sucrose when administered alone prevented the increase in motivation by the effective dose of olanzapine. These data show that increased motivation for palatable food is likely to be a significant contributor to olanzapine-induced weight gain. Moreover, the ability of sibutramine to reduce this motivation for palatable food may play an important role in the efficacy of sibutramine as an add-on treatment to counteract olanzapine-induced weight gain.     

The benzodiazepine partial agonists, Ro16-6028 and Ro17-1812, increase palatable food consumption in nondeprived rats

Two novel imidazobenzodiazepines, Ro16-6028 and Ro17-1812, have been described recently as partial agonists acting at benzodiazepine receptors. In a test of palatable food consumption using nondeprived rats, Ro16-6028 (0.01-10 mg/kg) and Ro17-1812 (0.01-10 mg/kg) were shown to produce dose-dependent increases in food intake. Ro16-6028 was more potent than Ro17-1812. Suriclone, midazolam, and the beta-carbolines ZK 93423 and ZK 91296 also significantly increased food intake. The maximum effects of Ro16-6028 and Ro17-1812 were at least equivalent to those obtained with full agonists acting at benzodiazepine sites. Neither Ro16-6028 nor Ro17-1812 reduced locomotion or rearing frequency in an open field test, although there was a reduction in grooming frequency. In contrast, the full agonist midazolam dose-dependently reduced all measures of general activity. The results indicate that some novel benzodiazepine partial agonists strongly stimulate food intake in the absence of side effects typical of the classical benzodiazepines.     

Modulation of morphine-induced antinociception by palatable solutions in male and female rats

The analgesic potency of opioid drugs varies as a function of gender, and can be modified by the intake of palatable sweet-tasting solutions. To determine if gender interacts with diet-induced changes in antinociceptive responses, male and female Long-Evans rats were fed laboratory chow and water alone, or chow, water and either a 32% w/v sucrose solution or a 0.15% w/v saccharin solution, and tested in two analgesic paradigms, the tail-flick test and the hot-plate test. For both tests, antinociceptive responses of male and female rats were tested following administration of cumulative doses (1.25, 2.5, 5.0, and 10.0 mg/kg, SC) of morphine sulfate. On the tail-flick test, morphine produced dose-related increases in antinociceptive responses. In addition, relative to both the chow only and saccharin conditions, chronic intake of the sucrose solution access significantly augmented morphine's antinociceptive properties. On the hot-plate test, when the plate was heated to 51 degrees C, morphine led to significant dose-related increases in antinociceptive responses, but diet did not affected antinociceptive responses. When the temperature of the hot plate was increased to 53 degrees C, there was a trend for animals given sucrose to have greater antinociceptive responses than those given either chow alone or saccharin. No differences in baseline pain sensitivity or morphine-induced analgesia were observed as a function of gender.     

Behavioural profile of exendin-4/naltrexone dose combinations in male rats during tests of palatable food consumption

Rationale

The glucagon-like peptide 1 receptor (GLP-1R) agonist exendin-4 potently suppresses food intake in animals and humans. However, little is known about the behavioural specificity of this effect either when administered alone or when co-administered with another anorectic agent.

Objectives

The present study characterises the effects of exendin-4, both alone and in combination with naltrexone, on behaviours displayed by male rats during tests with palatable mash.

Methods

Experiment 1 examined the dose-response effects of exendin-4 (0.025–2.5 μg/kg, IP), while experiment 2 profiled the effects of low-dose combinations of the peptide (0.025 and 0.25 μg/kg) and naltrexone (0.1 mg/kg).

Results

In experiment 1, exendin-4 dose dependently suppressed food intake as well as the frequency and rate of eating. However, these effects were accompanied by dose-dependent reductions in all active behaviours and, at 2.5 μg/kg, a large increase in resting and disruption of the behavioural satiety sequence (BSS). In experiment 2, while exendin-4 (0.25 μg/kg) and naltrexone each produced a significant reduction in intake and feeding behaviour (plus an acceleration in the BSS), co-treatment failed to produce stronger effects than those seen in response to either compound alone.

Conclusion

Similarities between the behavioural signature of exendin-4 and that previously reported for the emetic agent lithium chloride would suggest that exendin-4 anorexia is related to the aversive effects of the peptide. Furthermore, as low-dose combinations of the peptide with naltrexone failed to produce an additive/synergistic anorectic effect, this particular co-treatment strategy would not appear to have therapeutic significance.     

Effects of a chronic administration of two benzodiazepines on food intake in rats given a highly palatable diet

Chronic administration of benzodiazepines is known to increase food intake in numerous species. But this effect has been studied only after a unique daily injection and over a short part of the 24 hr cycle. In the present study, during 28 days, drugs were administered to rats receiving ordinary chow or a highly palatable diet (cafeteria diet): diazepam (DZ) (2.5 mg/kg IP) twice a day, or brotizolam (BR) (1 mg/kg IP), a longer acting compound, once a day. In the chow fed rats, DZ and BR provoked a post injection hyperphagia throughout the study, followed by a compensatory hypophagia resulting in 24 hr food intakes not different from those of controls; conversely neither body weight nor weight of fat pads were increased. The cafeteria diet provoked hyperphagia and overweight. DZ did not induce any supplementary hyperphagia. BR provoked a post injection hyperphagia, also compensated in time, resulting again in 24 hr food intakes, body weight gains and weight of fat pads not increased compared to those of cafeteria controls. Thus in the rat, benzodiazepine treatment increases food intake, but only acutely, and does not provoke any trend toward obesity.     

Ethanol's antiseizure efficacy is reduced by stress.

The ability of ethanol to antagonize the electrical precipitation of seizures in an incremental electroconvulsive shock paradigm was examined in groups of stressed and control mice. In stressed mice, the dose-response curve for ethanol's antiseizure efficacy was down-shifted and right-shifted relative to controls. These data may have clinical implications with respect to the interaction between stress, ethanol, and proneness to seizures.     

Ventricular fibrillation is reduced in hypothyroid rats with enhanced myocardial alpha-adrenoceptor responsiveness.

1. The severity of ventricular arrhythmias induced by coronary artery occlusion and reperfusion has been examined in control rats and animals made hypothyroid by pretreatment with 6-propylthiouracil (PTU). The maximal driving frequency and sensitivity of isolated left atria and papillary muscles to isoprenaline and to phenylephrine in the presence of propranolol, were also examined in tissues from control and hypothyroid animals. 2. Pretreatment with PTU resulted in a potentiation of responses to the alpha-adrenoceptor agonist phenylephrine in both left atria and papillary muscles, while responses to isoprenaline were depressed in left atria but unaltered in papillary muscles from hypothyroid animals. 3. In rats subject to coronary artery occlusion, PTU pretreatment reduced the incidence of ventricular fibrillation during acute myocardial ischaemia and abolished reperfusion-induced ventricular fibrillation. Mortality during myocardial ischaemia and reperfusion was also abolished. Diastolic blood pressure was similar in hypothyroid and control animals, but there was a small reduction in systolic blood pressure and a marked decrease in heart rate in PTU pretreated animals. 4. These results demonstrate that PTU-induced hypothyroidism represents a condition where cardiac alpha-adrenoceptor-mediated responses are enhanced but the severity of ischaemia- and reperfusion-induced arrhythmias is reduced.     

Scopolamine-induced suppression of paradoxical sleep is reversed by the somatostatin analogue SMS 201-995 in rats

The intraperitoneal administration of the octapeptide somatostatin analogue SMS 201-995 produced a significant increase in paradoxical sleep (PS) in rats. The suppression of PS by the muscarinic receptor blocker scopolamine was reversed by SMS 201-995. These findings confirm previous results demonstrating a role of somatostatin in the generation of PS. In addition they suggest that the suppression of PS by scopolamine may be due to an inhibitory effect on somatostatin release, rather than to an alteration of cholinergic function alone.     

Control of the dispersing process and pharmacokinetics in rats for lipid A analogue, E5531.

E5531 is a synthetic disaccharide analogue of lipid A which has a low toxicity but retains the ability to reduce production of tumour necrosis factor. This analogue has potential for use in the treatment of septic shock. An injectable formulation of E5531 would be useful, but dispersion in aqueous solution is a problem. In the present study the dispersing process for E5531 was evaluated using the pH-jump method (pH 11.0-->7.3). The size of the aggregates was decreased (reaching 20 nm) with increasing dispersing time in 0.003 M NaOH (pH 11.0). The membrane fluidity of the aggregates increased with increasing dispersing time. When prepared by the normal dilution method (pH 7.3-->7.3), the size of the aggregates remained constant at 140 nm and the membrane fluidity was smaller than that of samples prepared by the pH-jump method. This indicates that during dispersing at basic pH, the hydration proceeded in a normal manner, but then stopped, just after adjustment of the pH to 7.3. This suggests that the degree of hydration of the membrane is dependent on the dispersing time at pH 11.0. Using samples with different degrees of hydration and different membrane fluidity prepared by the pH-jump method, the pharmacokinetics and stability of the aggregates were evaluated after intravenous injection into rats. The data thus obtained confirmed that the membrane fluidity was correlated with the pharmacokinetics and stability in rat plasma. It was concluded that the pharmacokinetics of E5531 in rats can be controlled by changing the degree of hydration and membrane fluidity by means of using different dispersing times in alkaline solution (pH 11.0).     

Operant behavior to obtain palatable food modifies neuronal plasticity in the brain reward circuit

Palatability enhances food intake by hedonic mechanisms that prevail over caloric necessities. Different studies have demonstrated the role of endogenous cannabinoids in the mesocorticolimbic system in controlling food hedonic value and consumption. We hypothesize that the endogenous cannabinoid system could also be involved in the development of food-induced behavioral alterations, such as food-seeking and binge-eating, by a mechanism that requires neuroplastic changes in the brain reward pathway. For this purpose, we evaluated the role of the CB₁ cannabinoid receptor (CB₁-R) in the behavioral and neuroplastic changes induced by operant training for standard, highly caloric or highly palatable isocaloric food using different genetics, viral and pharmacological approaches. Neuroplasticity was evaluated by measuring changes in dendritic spine density in neurons previously labeled with the dye DiI. Only operant training to obtain highly palatable isocaloric food induced neuroplastic changes in neurons of the nucleus accumbens shell and prefrontal cortex that were associated to changes in food-seeking behavior. These behavioral and neuroplastic modifications induced by highly palatable isocaloric food were dependent on the activity of the CB₁-R. Neuroplastic changes induced by highly palatable isocaloric food are similar to those produced by some drugs of abuse and may be crucial in the alteration of food-seeking behavior leading to overweight and obesity.     

Stress-induced hyperthermia is reduced by rapid-acting anxiolytic drugs independent of injection stress in rats

Background

Stress-induced hyperthermia (SIH) is the transient rise in body temperature after encountering a stressor. The SIH response can be blocked by administration of various anxiolytic drugs prior to inducing stress. However, a drug injection involves handling and injection stress and therefore induces a SIH response itself. In the standard SIH test, drugs are therefore injected 60 min before stress induction to allow injection-induced hyperthermia to decline. This makes it difficult to study putative anxiolytic compounds with a short half-life. The present study therefore aimed to compare the effects of standard (stressful) and stress-free anxiolytic drug administration on the subsequent SIH response with a 10-minute injection-stressor interval.

Methods

Anxiolytic drugs with short half-lives (midazolam, 8-OH-DPAT, nicotine) were injected subcutaneously in rats using either a stressful (manual injection) or stress-free injection (subcutaneous cannula) method 10 min before novel cage stress. Body temperature and locomotor activity were measured using telemetric transmitters.

Results

Stressful and stress-free drug administration resulted in comparable drug effects on the stress-induced hyperthermia and locomotor responses in rats.

Conclusion

The present study shows that both stressful and stress-free drug injection shortly before a stressor results in reproducible attenuation of the SIH response in rats. In rats, a short injection-stressor interval can therefore be applied using the SIH model, enabling the study of putative anxiolytic drugs with short half-lives.     

Operant behavior to obtain palatable food modifies ERK activity in the brain reward circuit

Food palatability produces behavioral modifications that resemble those induced by drugs of abuse. Palatability-induced behavioral changes require both, the activation of the endogenous cannabinoid system, and changes in structural plasticity in neurons of the brain reward pathway. The ERK intracellular pathway is activated by CB₁ receptors (CB₁-R) and plays a crucial role in neuroplasticity. We investigated the activation of the ERK signaling cascade in the mesocorticolimbic system induced by operant training to obtain highly palatable isocaloric food and the involvement of the CB₁-R in these responses. Using immunofluorescence techniques, we analyzed changes in ERK intracellular pathway activation in the mesocorticolimbic system of wild-type and CB₁ knockout mice (CB₁?/?) trained on an operant paradigm to obtain standard, highly caloric or highly palatable isocaloric food. Operant training for highly palatable isocaloric food, but not for standard or highly caloric food, produced a robust activation of the ERK signaling cascade in the same brain areas where this training modified structural plasticity. These changes induced by the operant training were absent in CB₁?/?. We can conclude that the activation of the ERK pathway is associated to the neuroplasticity induced by operant training for highly palatable isocaloric food and might be involved in CB₁-R mediated alterations in behavior and structural plasticity.