The cholecystokinin-A receptor mediates inhibition of food intake yet is not essential for the maintenance of body weight.

Food intake and body weight are determined by a complex interaction of regulatory pathways. To elucidate the contribution of the endogenous peptide cholecystokinin, mice lacking functional cholecystokinin-A receptors were generated by targeted gene disruption. To explore the role of the cholecystokinin-A receptor in mediating satiety, food intake of cholecystokinin-A receptor-/- mice was compared with the corresponding intakes of wild-type animals and mice lacking the other known cholecystokinin receptor subtype, cholecystokinin-B/gastrin. Intraperitoneal administration of cholecystokinin failed to decrease food intake in mice lacking cholecystokinin-A receptors. In contrast, cholecystokinin diminished food intake by up to 90% in wild-type and cholecystokinin-B/gastrin receptor-/- mice. Together, these findings indicate that cholecystokinin-induced inhibition of food intake is mediated by the cholecystokinin-A receptor. To explore the long-term consequences of either cholecystokinin-A or cholecystokinin-B/gastrin receptor absence, body weight as a function of age was compared between freely fed wild-type and mutant animals. Both cholecystokinin-A and cholecystokinin-B/gastrin receptor-/- mice maintained normal body weight well into adult life. In addition, each of the two receptor-/- strains had normal pancreatic morphology and were normoglycemic. Our results suggest that although cholecystokinin plays a role in the short-term inhibition of food intake, this pathway is not essential for the long-term maintenance of body weight.

Wheel running attenuates the antinociceptive properties of morphine and its metabolite, morphine-6-glucuronide, in rats.

Recent work has shown that chronic exercise is associated with a reduction in the pain-relieving actions of opioid drugs in experimental animals. To determine whether this reduction represents an interaction between exogenously administered opioids and the endogenous opioid system, or is the result of altered drug pharmacokinetics, the antinociceptive actions of morphine and its metabolite, morphine-6-glucuronide (M6G), were compared in active and inactive female Long-Evans rats. Active animals were housed in running wheels and inactive animals in standard laboratory cages for 3 weeks preceding determinations of antinociception using the tail-flick test. At the end of the 3-week period, active rats were running the equivalent of 9-11 km a day. Antinociceptive responses, determined following subcutaneous injections of either morphine (0.625-20 mg/kg) or M6G (0.3-10.0 mg/kg), were significantly reduced in active rats relative to inactive rats. This reduction was manifested by both a lower magnitude of antinociception, and a shorter duration of antinociception after drug administration in active compared to inactive rats. This reduction was not associated with alterations in the estrous cycle or with differences in body weight between the active and inactive animals. The present results support the hypothesis that cross-tolerance develops between endogenous opioid peptides released in response to exercise and exogenously administered opioid drugs.

Intake of dietary sucrose or fat reduces amphetamine drinking in rats.

The effects of intake of a palatable food source on oral amphetamine intake were assessed in adult male Long-Evans rats. In Experiment 1, six rats were given an amphetamine sulfate solution (0.1 mg/ml) and four rats were given water as their sole source of fluid. Rats were given a choice of chow and granulated sucrose for a week, alternated with weeks when only chow was fed. In Experiment 2, eight rats were given the amphetamine solution, and four rats water to drink. Rats were fed chow and hydrogenated vegetable fat for a week alternated with weeks when only chow was available. In both experiments, rats drank significantly less of the amphetamine solution when the palatable food choice was available than when given only chow to eat. Intake of palatable foods had a significantly smaller effect on water intake. In both experiments, rats drinking the amphetamine solution took in less fluid and less calories and gained less weight than rats drinking water. However, in Experiment 1, when sucrose was available, rats drinking amphetamine consumed a significantly greater proportion of their calories as sucrose than rats drinking water. Similarly, in Experiment 2, rats drinking the amphetamine solution chose a significantly greater percentage of their calories as fat than rats drinking water. These results demonstrate that intake of sucrose or fat leads to a significant reduction in amphetamine intake, and that the anorectic effects of amphetamine are not equivalent for different types of foods.

Prior exposure to palatable solutions enhances the effects of naltrexone on food intake in rats.

Previous research has suggested that chronic intake of palatable foods and fluids enhances the activity of the endogenous opioid system. To examine this suggestion, the effect of naltrexone on food intake was examined in male Long-Evans rats with or without prior exposure to palatable solutions. In Experiment 1, rats were fed laboratory chow alone or laboratory chow and a 32% sucrose solution, and in Experiment 2, were fed chow alone, chow and a 32% Polycose solution, or chow and a 0.15% saccharin solution for three weeks. The sucrose, Polycose, and saccharin solutions were removed 18 h prior to drug administration. Rats then received injections of naltrexone hydrochloride (0.0, 0.3 or 3.0 mg/kg. sc) and chow intakes were measured during the subsequent 1, 2, 4, 6 and 24 h. Naltrexone injections had minimal effects on intakes of animals which previously had consumed only chow. In contrast, naltrexone led to significant dose-related decreases in chow intakes in rats which had previously consumed the sucrose, Polycose, or saccharin solutions. These results provide confirmation for the suggestion that chronic intake of palatable solutions alters the activity of the endogenous opioid system.

Chronic running-wheel activity decreases sensitivity to morphine-induced analgesia in male and female rats.

The effects of exercise on morphine-induced analgesia were examined in male and female Long-Evans rats. In Experiment 1, 10 male rats were housed in standard laboratory cages, and 10 in activity wheels for 20 days prior to nociceptive testing. Pain thresholds were assessed using a tail-flick (TF) procedure. Morphine sulfate was administered using a cumulative dosing procedure (2.5, 5.0, 7.5, 10.0, 12.5, and 15.0 mg/kg). TF latencies were measured immediately prior to and 30 min following each injection. In Experiment 2, morphine-induced analgesia was examined in females in an identical manner to that of Experiment 1. Additionally, to determine if the attenuation of morphine-induced analgesia was permanent or reversible, after the initial test nociceptive test, previously active female rats were placed in standard cages, and previously inactive females placed in running wheels for 17 days prior to a second nociceptive test. Baseline TF latencies were significantly shorter in active male rats than in inactive animals. Additionally, both active male and female rats displayed decreased morphine-induced analgesia relative to inactive controls. Moreover, females that had been inactive and then were permitted to run showed a suppression in morphine-induced analgesia relative to presently inactive rats, and to their own nociceptive responses when sedentary. In contrast, morphine-induced analgesia in initially active females who were housed in standard cages during part 2 of Experiment 2 was enhanced relative to their first nociceptive test and to presently active rats. Experiment 3 examined the effects of short-term (24 h) running on antinociception. Baseline TF latencies were shorter in active rats than inactive rats. However, no differences in morphine-induced analgesia were observed as a function of short-term exposure to exercise. Experiment 4 investigated whether differences in body weight contributed to the differences in morphine-induced analgesia between chronically active and inactive animals. %MPEs did not vary among male rats maintained at 100, 85, or 77% of their free-feeding body weight. These results indicate that chronic activity can decrease morphine's analgesic properties. These effects may be due to crosstolerance between endogenous opioid peptides released during exercise and exogenous opioids.

Altering dietary levels of protein or vitamins and minerals does not modify morphine-induced analgesia in male rats.

Previous research has demonstrated that chronic intake of nutritive sweet solutions, but not nonnutritive sweet solutions, enhances morphine's analgesic potency. To separate out the effects of sweet taste from other changes in dietary intake, which result when rats consume a sucrose solution, the effects of altering dietary levels of protein, or vitamins and minerals on morphine-induced analgesia were examined. In Experiment 1, 40 male Long-Evans rats were fed standard chow or a semipurified diet containing either 10, 20, or 40% protein. Three weeks later, antinociceptive responses to morphine were examined using the tail flick procedure. Tail flick latencies were measured immediately prior to and 30, 60, and 90 min after the administration of morphine sulfate (0.0, 1.25, 2.5, and 5.0 mg/kg, SC). At all three measurement times, antinociceptive responses increased directly as a function of the dose of morphine, but did not differ as a function of diet. In Experiment 2, 24 rats were maintained on either standard laboratory chow or semipurified diets containing 20% protein and either 100% or 25% of the recommended levels of vitamins and minerals for 3 weeks. Tail flick latencies were measured immediately prior to and 30 min after injections (SC) of 2.5 mg/kg morphine sulfate. This procedure was repeated until a cumulative dose of 10.0 mg/kg was obtained. Tail flick latencies increased significantly as a function of drug dose, but did not differ across dietary conditions. These results demonstrate that the increase in morphine-induced analgesia seen in rats consuming a sucrose solution is not due to alterations in either protein or micronutrient intake.

A role for melanin-concentrating hormone in the central regulation of feeding behaviour.

The hypothalamus plays a central role in the integrated regulation of energy homeostasis and body weight, and a number of hypothalamic neuropeptides, such as neuropeptide Y (ref. 1), galanin, CRH (ref. 3) and GLP-1 (ref. 4), have been implicated in the mediation of these effects. To discover new hypothalmic peptides involved in the regulation of body weight, we used differential display polymerase chain reaction to identify messenger RNAs that are differentially expressed in the hypothalamus of ob/+ compared with ob/ob C57B1/6J mice. We show here that one mRNA that is overexpressed in the hypothalamus of ob/ob mice encodes the neuropeptide melanin-concentrating hormone (MCH). Fasting further increased expression of MCH mRNA in both normal and obese animals. Neurons containing MCH are located in the zona incerta and in the lateral hypothalamus. These areas are involved in regulation of ingestive behaviour, but the role of MCH in mammalian physiology is unknown. To determine whether MCH is involved in the regulation of feeding, we injected MCH into the lateral ventricles of rats and found that their food consumption increased. These findings suggest that MCH participates in the hypothalamic regulation of body weight.