Increased fat consumption induced by morphine administration in rats

Patterns of caloric intakes and dietary self-selection of the three macronutrients, protein, fat and carbohydrate were examined in male rats following the administration of morphine sulfate (0.0, 1.0, 10.0 and 20.0 mg/kg, IP). Animals were given access to either ground Purina Chow or one of two dietary self-selection regimes, one with a high-fat ration (7.8 kcal/g) and the other with a fat ration isocaloric to the carbohydrate and protein rations (3.8 kcal/g). Animals received morphine injections at the beginning of a six-hour feeding period and nutrient intakes were measured at 1,2,4 and 6 hours postinjection. Similar patterns of macronutrient choice were observed for both animals maintained on the high-fat regime and animals with access to the isocaloric components following morphine injections. As a function of morphine injections, animals on both self-selection regimes increased fat intake while suppressing carbohydrate intake and exhibiting little modifications in protein intake.     

Differential effects of amphetamine and fenfluramine on dietary self-selection in rats

Daily caloric intakes and dietary self-selection of the three macronutrients, protein, fat and carbohydrate were examined in female rats following administration of d-amphetamine sulfate (0.0, 0.5, 1.0 and 2.0 mg/kg, IP) or fenfluramine hydrochloride (0.0, 1.5, 3.0 and 6.0 mg/kg, IP). Animals were maintained on ground Purina Chow or one of two self-selection regimes, one with a high-caloric fat ration (7.85 kcal/g) and the other with a fat ration isocaloric to the carbohydrate and protein rations (3.76 kcal/g). Animals received drug injections at the beginning of a daily 8-hour feeding period with nutrient intakes measured at 2, 4 and 8 hrs following injections. While both amphetamine and fenfluramine led to dose-related decreases in total caloric intakes, the two drugs resulted in different temporal patterns of feeding. Amphetamine produced its greatest effect on caloric intake during the first 2 hours of the feeding period, whereas fenfluramine suppressed caloric intake equivalently across the 8-hour feeding period. The two anorectic drugs also led to different patterns of nutrient choice. When animals were given the high-caloric fat ration, amphetamine selectively decreased fat intake while fenfluramine produced decreases in both protein and fat intakes, sparing carbohydrate intake. In contrast, when animals were given the isocaloric fat ration, amphetamine resulted in a general suppression of nutrient intakes while fenfluramine led to a sustained decrease in fat intake with a relative sparing of protein and carbohydrate consumption.     

Modifications of nutrient selection induced by naloxone in rats

Total caloric intake and dietary self-selection of the three macronutrients protein, fat, and carbohydrate were examined in male rats maintained on a 6-h feeding schedule following the administration of the opioid antagonist naloxone HCl (0.1, 1.0, and 10.0 mg/kg IP). Total caloric intake (calculated as the sum of caloric intakes from each of the macronutrients) was decreased for up to 2 h following naloxone administration. By the end of the 6-h feeding period, however, no differences in total caloric intakes were observed as a function of naloxone injections. Examination of intakes of the individual macronutrients revealed that naloxone differentially affected fat, carbohydrate, and protein consumption. Across the 6-h feeding period, animals consumed less calories from the fat ration following all three doses of naloxone than after saline injections. Carbohydrate intake was decreased for up to 2 h following naloxone injections, but returned to control values by the end of the 6-h feeding period. Protein intake, in contrast to fat and carbohydrate intakes, did not vary as a function of naloxone administration. Results of the present experiment are contrasted with patterns of dietary self-selection observed following morphine administration.     

Peripheral serotonin administration selectively reduces fat intake in rats

Recent research has led to the hypothesis that serotonergic mechanisms may be involved in both the control of energy intake and appetites for specific nutrients. Most of this research has focused on serotonin (5-HT) within the central nervous system. However, there is evidence which suggests that peripheral 5-HT also may be involved in the control of energy intake and nutrient selection. To further assess this suggestion, the effects of peripheral 5-HT administration on energy consumption and nutrient intakes were examined in adult male Sprague-Dawley rats given separate sources of protein, fat and carbohydrate. Administration of 5-HT (doses ranging from 2-6 mg/kg) led to significant dose-related decreases in total energy intake in both freely feeding and food-restricted rats. Examination of individual nutrient intakes revealed that following 5-HT, fat intake was more suppressed than either carbohydrate or protein intakes. Administration of the 5-HT antagonist, methysergide, blocked the suppressive effects of 5-HT on both total energy intake and fat intake. The present data support the proposal that peripheral serotonergic mechanisms play a role in ingestive behaviors.     

Naltrexone does not prevent acquisition or expression of flavor preferences conditioned by fructose in rats

The effects of the general opioid antagonist, naltrexone, on the acquisition and expression of flavor preferences conditioned by the sweet taste of fructose were examined. Food-restricted rats were trained over eight daily alternating one-bottle sessions (2 h) to drink an 8% fructose solution containing one novel flavor (CS+/F) and a less preferred 0.2% saccharin solution containing a different flavor (CS-/S). Four groups of rats were treated daily with either saline (control group) or naltrexone doses of 0.1, 1.0, or 5.0 mg/kg during training. Preferences were assessed in two-bottle tests with the CS+/S and CS-/S flavors presented in 0.2% saccharin solutions following saline injections. Naltrexone dose-dependently reduced fructose and saccharin intakes during training, confirming the drug's well-known suppressive effect on the intake of sweet solutions. Despite their reduced training intakes, the naltrexone groups displayed preferences for the CS+/S over the CS-/S (72-86%) that were similar to that of the control group (78%). The effect of naltrexone on the expression of the CS+/S flavor preference was evaluated by treating control rats with naltrexone (0.1-5 mg/kg) prior to CS+/S vs. CS-/S choice tests. The drug doses produced a dose-dependent reduction in CS+/S intake but did not significantly attenuate the CS+/S preference. These data are consistent with the relative inability of naltrexone to reduce flavor-flavor conditioning by sucrose in sham-feeding rats and flavor-nutrient conditioning in rats receiving intragastric sucrose infusions. In contrast, dopamine antagonists reduce both sucrose- and fructose-conditioned flavor preferences, which indicates the sensitivity of these conditioning paradigms to neuropharmacological manipulations. These data indicate that the endogenous opioid system, unlike the dopamine system, does not play a major role in either the acquisition or expression of flavor preference learning as measured in two-bottle choice tests.     

An opioid receptor antagonist, naltrexone, does not alter taste and smell responses in humans

Several studies have shown that an opioid receptor antagonist, naltrexone, decreases palatable food consumption. Naltrexone has also been reported to reduce ethanol intake in alcohol-preferring rodents and human alcoholics. The aim of the present study was to assess the effects of naltrexone on taste and smell responses in healthy male volunteers. Naltrexone did not alter intensity and pleasantness of sucrose, quinine, citric acid, sodium chloride, and ethanol taste. Similarly, ratings of olfactory stimuli (orange extract and ethanol) and Coca-Cola flavor were not influenced by the opioid antagonist. Our findings may indicate that: (i) naltrexone exerts marginal, if any, effects on gustatory and olfactory responses in humans; (ii) the drug does not alter orosensory responses to ethanol.     

Naltrexone decreases D-amphetamine and ethanol self-administration in rhesus monkeys

Amphetamines are the second most highly abused illicit drugs worldwide, yet there is no pharmacological treatment for amphetamine abuse and dependence. Preclinical studies and, more recently, human studies, suggest that the opioid receptor antagonist, naltrexone, might be useful in the treatment of amphetamine abuse. Naltrexone, an opioid receptor antagonist, is currently used for the treatment of alcohol dependence. The aim of this study was to explore the ability of naltrexone to modify self-administration of amphetamine or ethanol in rhesus monkeys. Monkeys were trained to respond to intravenous injections of either D-amphetamine (0.003 mg/kg/injection) or ethanol (0.05 g/kg/injection) on a fixed ratio 30 schedule. Naltrexone (0.01-1 mg/kg) was administered intramuscularly 30 min before the start of treatment test sessions. Naltrexone dose-dependently decreased both amphetamine and ethanol self-administration. These findings support the potential use of naltrexone as therapy for amphetamine and polydrug abuse.     

Naltrexone fails to block the acquisition or expression of a flavor preference conditioned by intragastric carbohydrate infusions

The effects of naltrexone on the expression and acquisition of flavor preferences conditioned by the postingestive actions of carbohydrates were investigated. Food-restricted rats (Experiment 1) were given one-bottle training with one flavored saccharin solution (CS+) paired with intragastric (IG) infusions of 16% sucrose, and another flavored saccharin solution (CS-) paired with water infusions. In two-bottle tests CS+ was preferred to CS-, and naltrexone (1.0-5.0 mg/kg) reduced total intake but not CS+ preference. In Experiment 2 food-restricted rats that received naltrexone (0.1 or 1.0 mg/kg; NTX group) throughout one-bottle training consumed less CS+ and CS- than did saline-treated control rats. Yet, the NTX and control groups displayed similar CS+ preferences during two-bottle tests when treated with saline or naltrexone (0.1-5.0 mg/kg). In Experiment 3, rats were trained to accept more CS+ than CS- in one-bottle tests. Naltrexone (0.1-2.5 mg/kg) reduced the one-bottle intakes of both solutions, and the rats continued to consume more CS+ than CS-. We conclude that the opioid system modulates the consumption of flavored solutions, but is not critically involved in the acquisition or expression of flavor preferences conditioned by IG carbohydrate.     

Increased carbohydrate consumption by rats as a function of 2-deoxy-D-glucose administration

Dietary self-selection was examined following the administration of the glucoprivic agent, 2-deoxy-D-glucose (2-DG), in adult male rats given access to separate sources of the three macronutrients, protein, fat and carbohydrate. All animals received injections (IP) of saline, 250, 500 and 750 mg/kg 2-DG with nutrient intakes measured at 2, 4, 6 and 24 hrs following injections. Animals consumed significantly more carbohydrate at 4, 6 and 24 hrs after injections of 500 and 750 mg/kg 2-DG than after saline injections. In contrast, fat intake was significantly suppressed by all three doses of 2-DG at 2 hr, by 250 and 750 mg/kg 2-DG at 4 and 6 hrs, and by 750 mg/kg 2-DG at 24 hr after injections. Protein intake was significantly decreased by all three doses of 2-DG at 2 hr after injections. As a result of the increase in carbohydrate intake and complimentary decrease in fat intake following 2-DG injections, total caloric intake of animals given the self-selection regime was not modified as a function of drug administration. In comparison, rats given a single nutritionally complete diet (ground Purina Laboratory Chow) consumed significantly more calories following 2-DG administration than following saline injections. The ability of animals to make appropriate modifications in nutrient selection following regulatory challenges is discussed.     

Baclofen, raclopride, and naltrexone differentially affect intake of fat/sucrose mixtures under limited access conditions

This study assessed the effects of the opioid antagonist naltrexone, the dopamine 2-like (D2) antagonist raclopride, and the GABA_B agonist baclofen on consumption of fat/sucrose mixtures (FSM) using a limited access protocol. Sixty male Sprague-Dawley rats were grouped according to two schedules of access (Daily [D] or Intermittent [I]) to an optional FSM. Each FSM was created by whipping 3.2% (L), 10% (M), or 32% (H) powdered sugar into 100% vegetable shortening in a w/w manner (n = 10 per group). One-hour intakes of the IL and IM groups were significantly greater than intakes of the respective DL and DM groups, thus fulfilling our operational definition of binge-type eating in these groups. Baclofen reduced intakes of the L and M mixtures regardless of access schedule, but failed to reduce intake of the H mixture. Naltrexone reduced intake in all groups, but potency was greater in IL rats than in DL rats. Furthermore, potency was attenuated in Intermittent rats, but enhanced in Daily rats, at higher sucrose concentrations. Raclopride reduced intake in the DL and stimulated intake in the IL groups, reduced intake in both M groups, and was without effect in both H groups. These results indicate that fat/sucrose mixtures containing relatively low concentrations of sucrose allow distinctions to be made between: 1) intakes stimulated by different access schedules and 2) opioid and dopaminergic modulation of those intakes. These results also suggest that brief bouts of food consumption involving fatty, sugar-rich foods may prove to be particularly resistant to pharmacological intervention.     

Pharmacology of sucrose-reinforced place-preference conditioning: effects of naltrexone

Two experiments investigated the role of the opioid system in sucrose-reinforced conditioned place preferences (CPPs) in rats. Experiment 1 examined the effects of a general opioid antagonist, naltrexone, on the expression of a CPP acquired in the absence of the drug. Subjects were trained to associate one compartment of a two-compartment chamber with sucrose and the other compartment with water. Rats displayed a preference for the sucrose-associated compartment in a choice test without sugar or water available following vehicle saline treatment. Naltrexone doses of 2.5 and 5.0 mg/kg reduced this preference for the sucrose-associated compartment. Experiment 2 examined the effects of naltrexone on the acquisition as well as the expression of CPPS. Different groups of rats received daily injections of either saline, 0.1, 1.0, or 5.0 mg/kg of naltrexone prior to each training session, and then these groups were given a choice test for the CPP after saline or naltrexone injections. Although naltrexone treatment attenuated the expression of CPPs in each group relative to saline treatment, there were no group differences during these tests in the magnitude of the preferences. Moreover, all groups displayed equal acquisition of CPPs despite the fact that naltrexone dose dependently decreased sucrose intake during the training phase. Together, the results indicate that the opioid system modulates the expression but not the acquisition of sucrose-reinforced CPPs.     

Reduction of fat and protein intakes but not carbohydrate intake following acute and chronic fluoxetine in female rats.

Fluoxetine hydrochloride, a selective serotonin reuptake inhibitor, leads to reductions in food intake and body weight and is under investigation as a possible treatment for obesity. Additionally, it has been suggested that fluoxetine administration could lead to a selective suppression in carbohydrate consumption. Because women more often than men seek weight reduction treatment, the present study examined the acute and chronic effects of fluoxetine on food intake, macronutrient selection, body weight, estrous cycle, and motor activity in female rats. Female Long-Evans rats were provided with separate sources of protein, fat and carbohydrate, and nutrient intakes were recorded following single (5.0, 10.0, and 20.0 mg/kg, IP) and chronic daily (10 mg/kg for 28 days) injections of fluoxetine. Acute and chronic administration of fluoxetine significantly reduced total caloric intake when compared to vehicle treatment. Moreover, fluoxetine significantly suppressed fat and protein intakes, but not carbohydrate intake following both acute and chronic drug administration. Animals chronically treated with fluoxetine gained significantly less weight than animals treated with vehicle. Chronic fluoxetine treatment did not significantly alter estrous cycle. However, in both fluoxetine- and vehicle-treated animals, total caloric intake, and carbohydrate and protein intakes were reduced and fat intake was increased when estrogen levels were high. Fluoxetine significantly reduced motor activity up to 4 h postinjection, and increased motor activity 24 h postinjection.     

Effects of oral loperamide on efficacy of naltrexone, baclofen and AM-251 in blocking ethanol self-administration in rats

Naltrexone is a μ-opioid receptor antagonist that has been extensively studied for its ability to block the rewarding effects of ethanol. Opioid receptors are widely distributed within the gastrointestinal tract (GIT). Typically, naltrexone is administered by parenteral routes in nonclinical studies. We initially tested if opioid receptors within the GIT would influence the ability of oral naltrexone to inhibit ethanol oral self-administration in rats using the co-administration of oral loperamide, a peripherally restricted opioid agonist. As expected, oral naltrexone only had modest effects on ethanol intake, and the response was not dose-dependent. However in rats, treatment with loperamide prior to the administration of naltrexone resulted in a suppression of ethanol intake which approached that observed with naltrexone given by the subcutaneous (SC) route. Importantly, administration of loperamide prior to administration of naltrexone did not alter blood concentrations of naltrexone. We then evaluated if oral loperamide would enhance effects of baclofen (a GABA_B receptor agonist) and AM-251 (a CB-1 receptor antagonist) and found that pre-treatment with loperamide did potentiate the action of both drugs to reduce ethanol self-administration. Finally, the specific opioid receptor type involved was investigated using selective μ- and κ-receptor antagonists to determine if these would affect the ability of the AM-251 and loperamide combination to block ethanol drinking behavior. The effect of loperamide was blocked by ALKS 37, a peripherally restricted μ-receptor antagonist. These data suggest an important role for opioid receptors within the GIT in modulating central reward pathways and may provide new insights into strategies for treating reward disorders, including drug dependency.     

Naltrexone, serotonin receptor subtype antagonists, and glucoprivic intake: 2. Insulin.

Opiate antagonist inhibition of deprivation-induced intake and 2-deoxy-D-glucose (2DG) hyperphagia is significantly enhanced by the 5-hydroxytryptamine3 (5-HT3) antagonist, ICS-205,930. Interactions between opiate antagonists and either 5-HT or 5-HT2 antagonists produced smaller effects. The present study evaluated whether insulin (5 U/kg) hyperphagia was affected by methysergide (0.5-5 mg/kg), ritanserin (0.25-2.5 mg/kg), and ICS-205,930 (0.5-5 mg/kg) alone or in combination with naltrexone (2.5-10 mg/kg). Whereas ICS-205,930 stimulated insulin hyperphagia across the 6-h time course, ritanserin and, to a lesser degree, methysergide reduced insulin hyperphagia. Naltrexone marginally (19-33%) reduced insulin hyperphagia. Pairing naltrexone with either ICS-205,930 or ritanserin significantly suppressed insulin hyperphagia after 2 h. Pairing naltrexone with each of the serotonin antagonists significantly enhanced insulin hyperphagia after 4 and 6 h. These data suggest that 5-HT2 and 5-HT3 receptor subtypes interact with opioid systems to modulate insulin hyperphagia. Given that central insulin reduces food intake and body weight, the interaction between serotonergic and opioid systems may occur peripherally.     

Naltrexone and morphine alter the discrimination and plasma levels of ethanol

The opioid antagonist, naltrexone, reduces intake of, and operant responding for, ethanol, but reports of how the opioid agonist morphine alters these effects are conflicting. We examined the discrimination and plasma levels of ethanol with naltrexone and morphine pretreatments. Rats were trained to discriminate ethanol (1.5 g/kg; i.g.) from water, under a two-lever, food-reinforced procedure. Ethanol and pentobarbital, but not amphetamine, substituted for ethanol in a dose-related manner. Naltrexone reduced ethanol-induced, ethanol-appropriate responding to about 35%, but the peripherally-acting antagonist, naltrexone methobromide, was without effect. Morphine neither substituted for nor enhanced ethanol-appropriate responding. Rather, ethanol-induced, ethanol-appropriate responding was attenuated in a dose-related manner by morphine administration. Neither naltrexone nor morphine altered ethanol-appropriate responding to the substitution with pentobarbital. In another group of rats, both naltrexone and morphine decreased plasma ethanol levels, and delayed the time of peak concentrations, suggesting that opiates alter the behavioral effects of ethanol through both pharmacokinetic and pharmacodynamic mechanisms. The similarities between an opioid agonist and an antagonist suggest that either naltrexone has opioid agonist-like effects, or that these effects occur through non-opioidergic mechanisms.     

Effects of naltrexone on food preference and concurrent behavioral responses in food-deprived rats

Naltrexone (0.05-5.0 mg/kg, SC) was administered to food-deprived rats prior to a 15-min food-preference test. Total food intake and feeding duration was reduced following administration of the opiate antagonist. However, while naltrexone reduced the consumption of the initially-preferred chocolate-coated cookies, the ingestion of the nonpreferred standard laboratory chow pellets was significantly enhanced. These data cannot be explained in terms of a general anorexic effect and nonspecific suppression of feeding responses. Instead, they indicate that naltrexone reduced preference for the highly palatable cookies, so that a feeding response to the chow pellets emerged. Under the conditions of test-familiarity, naltrexone did not reduce grooming, locomotion or rearing duration. An increase in locomotion may have been secondary to the reduction in feeding. The results agree with previous data from animal and human studies in suggesting that endogenous opioid peptide activity is involved in the palatability of preferred foods.     

Diet selection following a chronic morphine and naloxone regimen

Total caloric intake and patterns of dietary self-selection of the three macronutrients, protein, carbohydrate and fat, were examined in adult male rats maintained on a 6-hr feeding schedule following daily injections of morphine (10 mg/kg), naloxone (1 mg/kg), the two drugs together, and saline. Animals received drug injections for 10 consecutive days. All animals received saline injections for the 5 days preceding and 5 days following the experimental period. Naloxone injections led to a significant reduction in total caloric intake. Neither morphine nor morphine and naloxone together significantly affected total caloric intake. Each of the drugs had a distinct effect on macronutrient selection. Morphine produced a significant increase in fat intake and decrease in carbohydrate intake, while naloxone led to a slight reduction in fat intake. When the two drugs were given together, a significant elevation in carbohydrate intake and reduction in fat intake were observed. Protein intake was not affected by any of the drugs. These results are discussed with respect to the hypothesized role of the endogenous opioid system in the regulation of energy balance.     

Opioid modulation of thermal dehydration-induced thirst in rats.

Male Sprague-Dawley rats were utilized to study the effects of the opioid receptor antagonists, naloxone and naltrexone, on thirst induced by thermal dehydration. In an initial experiment, the depressant effect of naloxone (1.0 mg/kg, IP) on the water intake of rats deprived of water for 24 h was confirmed. In subsequent experiments, rats were thermally dehydrated by exposing them without water to a 40 degrees C environment for 1-4 h. Following heat exposure, rats were injected with either naloxone or naltrexone either IP or ICV. Fifteen minutes later, rats were provided with water and water intake was measured for 2 h. Both naloxone and naltrexone had dose (0.1-5.0 mg/kg, IP)-dependent effects of reducing water intake of rats thermally dehydrated for 3 h. Water intake of rats thermally dehydrated for 2 or 4 h was also attenuated by pretreatment with naloxone. Rats thermally dehydrated for 3 h exhibited decreases in water intake following ICV injection of either naloxone or naltrexone at a dose of 50 micrograms. Neither naloxone nor naltrexone had an effect on urine output in any experiment. The water intake data support the hypothesis that thirst induced by thermal dehydration in rats is modulated by an opioid mechanism.     

Chronic d-amphetamine inhibits opioid receptor antagonist-induced supersensitivity.

Chronic treatment with an opioid antagonist, such as naltrexone, increases opioid receptor density and opioid agonist potency. Since stimulants such as d-amphetamine can increase opioid potency and opioid abusers may administer stimulants during naltrexone treatment, the effect of chronic d-amphetamine on naltrexone-induced opioid receptor upregulation and supersensitivity was examined in mice. Mice were implanted s.c. with a 15 mg naltrexone or placebo pellet for 8 days. Mice were injected daily with saline or d-amphetamine (7.5 or 5.0 mg/kg per day s.c.) for 7 days beginning 24 h following implantation. Naltrexone and placebo pellets were removed on the 8th day, and 24 h later mice were tested for morphine analgesia (tail-flick) or whole brain was removed and opioid receptor binding studies were conducted. Chronic naltrexone significantly enhanced the analgesic potency of morphine in saline-treated mice. However, naltrexone treatment did not increase morphine potency in mice treated with d-amphetamine. In binding studies, naltrexone increased [3H][D-Ala2,NMePhe4,Gly-ol5]enkephalin (DAGO) Bmax (+60-70%) without altering KD in both saline- and d-amphetamine-treated mice. Results from studies with 2 nM [3H][D-Pen2,D-Pen5]enkephalin (DPDPE) were similar. These studies indicate that daily d-amphetamine can limit naltrexone-induced supersensitivity but not receptor upregulation. Thus, upregulation can be dissociated from functional supersensitivity.     

Norepinephrine, clonidine, and tricyclic antidepressants selectively stimulate carbohydrate ingestion through noradrenergic system of the paraventricular nucleus

Using a self-selection feeding procedure, the present experiments examined the impact of central and peripheral injection of the alpha-adrenergic agonist clonidine (CLON) and the tricyclic antidepressant drugs amitriptyline (AMIT) and chlorimipramine (CIMIP) on nutrient selection in the adult male rat. In tests with mixed diets or with separate sources of the 3 macronutrients (carbohydrate, protein, and fat) simultaneously available, the following results were obtained: Peripheral and paraventricular nucleus (PVN) injection of CLON stimulated total food intake and preferentially increased ingestion of carbohydrate. Little or no change in protein or fat intake was observed. This pattern of response is similar to that observed with norepinephrine. PVN injection of AMIT and peripheral injection of CIMIP also selectively enhanced carbohydrate intake. These drug effects on carbohydrate selection occurred under a variety of conditions, including with mixed diets and pure dietary nutrients; under ad lib and restricted feeding conditions; in short (1 hr) as well as long (6 hr) test intervals; and in the absence or presence of a change in total calorie intake. Based on this and other evidence, it is proposed that noradrenergic neurons innervating the PVN in the rat play a role in regulating carbohydrate selection, and that this neurochemical system mediates the stimulating action of CLON and antidepressants on carbohydrate ingestion.