Aspects of abstinence after morphine ingestion

Sprague-Dawley male rats were intoxicated with morphine, using an ingestion method where exposed and control rats received equivalent amounts of calories and nutrients. The degree of physical dependence on morphine was demonstrated by studying and quantifying abstinence symptoms after withdrawal or after administration of opiate antagonists. The aims of the study were (1) to further enlighten the specificity and validity of the intoxication method concerning physical dependence, and (2) to determine whether some of the abstinence signs might be of value to facilitate quantitation of the degree of physical dependence on morphine, with diet and fluid intake being maintained under control. Withdrawn rats showed a decreased fluid diet intake and a body weight loss, the latter partly due to anorexia. Other mild abstinence signs were irritation, tremor and some motor excitation. The body weight loss during the first day of morphine withdrawal was proportional to the accumulated drug dose (between 25 and 300 mg morphine PO/kg b.wt.). However, prolonged morphine treatment on one dose (340 mg/kg b.wt.) did not reinforce the body weight changes caused by morphine withdrawal. The succeeding weight gain some days after morphine withdrawal was not entirely dependent on the amount of fluid diet intake. Methadone was shown to partially block the decrease in diet intake and the weight loss seen during morphine withdrawal. The naloxone-precipitated withdrawal symptoms were motor excitation, cholinergic signs, body weight loss, diarrhoea and decreased diet intake. The weight loss 2 hr after naloxone administration to long-term intoxicated rats was proportional to the naloxone dose.(ABSTRACT TRUNCATED AT 250 WORDS)     

A study on drug dependence using fast acting drugs]

Physical dependence on narcotics is induced in laboratory animals by intermittent parenteral administration (2 approximately 3 times daily). However, inducing of dependence on pethidine has been unsuccessful using the parenteral method. Recently, it has been reported that physical dependence on pethidine can be induced by continuous infusion methods (5.6). In the present experiment, pethidine was administered to rats (n=5 approximately 6) by ingestion of pethidine-admixed food preparations (0.5 approximately 4.0 mg/g of feed). The results indicated that (a) when rats are allowed free access to two food preparations (0.5 mg/g vs. 1 mg/g of food) for 7 weeks, spontaneous intake ratios of food (1 mg/g of food) gradually increased from 15% to 30% after 3 weeks. (b) Abrupt withdrawal for 48 hr after a 10 day administration period (2 mg/g of food on day 1 approximately 3 and 4 mg/g of food on day 4 approximately 10) resulted in a loss of body weight in the next 24 hr, and the prewithdrawal level of body weight was recovered in 48 hr. (c) The time course of body weight and food intake during the first 24 hr withdrawal period demonstrated the characteristic pattern of abstinence syndrome of pethidine, viz. early onset (12 approximately 13 hr) and rapid recovery (within 48 hr), as compared to morphine withdrawal. (d) Suppression of pethidine abstinence of both a single injection of morphine (10 mg/kg, s.c.) and substitution for morphine-admixed food was also realized. (e) When levallorphan (5 mg/kg, s.c.) was administered to both pethidine and morphine dependence rats, the maximal decrease in body weight was less than that in morphine dependent rats. These data indicate that pethidine possesses about one fifth the dependence liability of morphine and the maximal abstinence syndrome appears within 24 hr after withdrawal. Conclusively, application of a drug-admixed food preparation in drug dependence tests in rats has proven to be a useful method, particularly in the case of pethidine-like drugs which rapidly disappear from the blood.     

Comparison of development of drug dependence in naive and drug dependence-experienced rats]

Phenobarbital, chordiazepoxide, diazepam and/or morphine were repeatedly administered to both male and female rats (N equals 10) for 4 similar to 6 weeks. The drug dose was gradully increased from 5 to 10, 20, 40, 80 and 160 mg/kg once daily (p.o) at seven day intervals. In the case of morphine, the last dose was 40 mg/kg. The drugs were constantly withdrawn for 24 hr at 8 day intervvals. None of the rats were given drugs for 16 days after administration of the last scheduled dose in order to recover their initial weight (Exp. I). Onset of dependence formation, decrease in body weight and food intake, days required to reach the maximum decrease in body weight and duration of withdrawal signs were observed throughout this experiment. The rats (drug dependence-experimented rats) who survived the first stage of this experiment were continuously subjected to re-administration by the same dosage schedule as in Exp. I (seven days of drug administration, 48hr of withdrawal). The re-administered rats showed a more rapid onset of dependence formation and a longer duration of decreas in boy weight during 16 days withdrawal than did the naive rats. It is concluded, that in addition to the decrease in body weight by withdrawal plus duration of the withdrawal signs, the onset of drug dependence formation is also a specific factor.     

Effect of propranolol on antinociceptive and withdrawal characteristics of morphine

Propranolol at a dose (10 mg/kg) which did not alter tail-flick latency by itself, did not alter the ED50 of morphine when given 10 min prior to the narcotic. Propranolol at doses of 10 and 25 mg/kg given 10 min prior to naloxone challenge did not significantly alter the frequency of naloxone induced jumping 72 hr after morphine pellet implantation. The ED50 of naloxone in morphine pelleted mice was not altered by treatment with propranolol at 0, 24, and 48 hr after pellet implantation. Naloxone caused hyperactivity in mice when administered 72 hr after morphine pellet implantation. An injection of 25 mg/kg propranolol 10 min prior to naloxone did not block this hyperactivity. In addition, administration of 10 mg/kg of propranolol every 8 hr to rats during withdrawal from morphine failed to alleviate the withdrawal syndrome as evidenced by changes in either body weight or water intake. These data suggest that the beta-adrenergic blocking agent, propranolol, does not alter the antinociceptive activity or lessen the withdrawal syndrome of morphine in rodents.     

Production of tolerance and physical dependence in the rat by simple administration of morphine in drinking water.

1 Rats are capable of consuming solutions of morphine sulphate in drinking water ad libitum in the absence of taste-masking chemicals and without the need for scheduled provision or prior parenteral administration of the drug. 2 The success of this method depends on the initial provision of a 0.1 mg/ml solution of morphine sulphate. 3 When the drug concentration is increased to 0.4 mg/ml, the rats achieve an average daily intake of 50 mg/kg body wt. each. 4 Daily intake of morphine may be increased by at least about three fold by increasing the drug concentration to 1.2 mg/ml. 5 Oral morphine administration causes only a moderate loss in body weight. 6 Rats whose daily intake of the drug is 50 mg/kg exhibit tolerance to the analgesic action of morphine and show a drastic loss in body weight at 24 h after withdrawal and most of the behavioural symptoms of the naloxone-precipitated withdrawal syndrome. 7 It is suggested that this simple method of morphine administration is suitable for further biochemical and behavioural studies of the actions of the drug.     

Dependence on and preference for morphine. (III) Improvement of spontaneous drug intake after combined treatment with morphine and cocaine]

One group of rats (n equals 6) was allowed free access to food combined with morphine and cocaine for 3 weeks, another group combined with either morphine or cocaine alone for 3 weeks. Intensity of decrease in body weight by withdrawal and ratios of spontaneous morphine and cocaine intake were compared, and the cross-spontaneous drug intake between morphine and cocaine using cocaine and morphine dependent rats was investigated, respectively. The results indicate that (a) decreased rate in body weight by withdrawal after 3 weeks administration with morphine-cocaine mixtures increased slightly, and spontaneous intake ratio for the higher drug-admixed food during administration period was also slightly increased as compared with a morphine added diet alone. (b) However, spontaneous intake ratio of both morphine [morphine (1 mg/g of food) vs. a normal diet (N.D.)] and cocaine [cocaine (2 mg/g of food) vs. N.D] increased 30 and 50 approximately 60% respectively, and these changes were maintained for about two months. (c) Spontaneous cocaine intake ratio after 3 weeks administration of morphine was almost the same level as that in cocaine dependent rats and decreased dependently when the administration period was extended. Spontaneous morphine intake ratio after 3 weeks administration of cocaine was 20% lower in total daily intake ratio as compared to morphine dependent rats, but spontaneous morphine intake ratio as compared to morphine dependent rats was gradually increased as in morphine dependent rats. (d) Furthermore, morphine-quinine combined treatment and cocaine or codeine treatment had no effect on spontaneous intake of quinine. These data suggest that combinations of morphine and cocaine have promoting properties of morphine and cocaine intake, respectively. Cross-spontaneous drug intake between morphine and cocaine was successful only in cocaine dependent rats using this method.     

Dietary modulation of the anorectic potency of amphetamine

The effects of intake of an amphetamine solution on food and fluid intakes, body weight (b.wt.) and feed efficiency (FE) were examined in rats fed either a high-carbohydrate (HC) (65% of total calories) or high-fat (HF) (65% of total calories) diet. During a 17-day predrug period, neither caloric intake, fluid intake, b.wt. nor feed efficiency (FE) differed as a function of diet. When given a 0.1 mg/ml amphetamine sulfate (AMPH) solution as their sole source of fluid, rats in both diet groups decreased fluid intake by an equivalent amount. While diet did not influence AMPH intake, it did alter the drug's effects on caloric intake, b.wt. and FE. In both diet groups, rats drinking AMPH decreased caloric intake, b.wt. gain and FE relative to rats which drank water. However, rats fed the HC diet decreased caloric intake less, but lost more weight than rats fed the HF diet. Further, rats fed the HC diet displayed a rapid tolerance to the anorectic effects of AMPH, with no tolerance to the drug's effect on b.wt. In contrast, rats fed the HF diet suppressed caloric intake throughout the drug period, but weighed more than rats fed the HC diet. Thus, when drinking the AMPH solution, FE of rats fed the HC diet was significantly lower than that of rats fed the HF diet. When the drug was removed, rats eating the HC diet failed to alter caloric intake and b.wt. In contrast, rats fed the HF diet increased caloric intake and gained weight. These data indicate that dietary factors must be considered when evaluating the anorectic actions of psychopharmacological agents.     

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.     

A new ingestion method for long-term morphine intoxication in rat

A new method for long-term morphine intoxication in rat was developed. It was designed to deal with the nutritional imbalance and body weight loss that generally occurs using conventional techniques for morphine treatment. The morphine is administered in a nutritionally complete diet. Also pair-feeding is used to deal with intoxicated rats that do not eat the same amount of food as controls. The technique was validated during the study of different intoxication conditions, using different initial doses, dose increments and final doses. An initial dose of 25 mg morphine/kg b.w., raised exponentially up to 340 mg/kg b.w. in 8 days, made the rats dependent, as tested by withdrawal signs, precipitated by excluding morphine from the diet, or by administration of antagonists. A final dose of up to 715 mg morphine/kg b.w. was reached in 13 days without decreased food intake. However, initial doses of 340 or 715 mg/kg led to impaired weight gain and diet consumption. Plasma morphine levels of 3 μg/ml serum were reached on a dose of 340 mg/kg b.w. Also, preference for morphine diet over control diet was evaluated by choice tests. The technique is simple, time-saving and inexpensive, allowing the treatment of numerous animals for long periods under standardized intoxication conditions. No animals get ill or die.     

The effect of neurotensin on food consumption in the rat

The effect of neurotensin on feeding behavior were studied in rats. Intracerebroventricular administration of neurotensin (3.3-30 micrograms) produced a dose-related decrease in food intake in 24 h food deprived rats. Acute intracerebroventricular injection of neurotensin (30 micrograms) shortly after the ingestion of a novel flavor did not produce a flavor aversion during testing 48 h later, suggesting that reduction of food intake by low doses of centrally administered neurotensin is not related to a conditioned taste aversion. Intracerebroventricularly administered thyrotropin-releasing hormone (2.2 micrograms) also inhibited food intake and appeared to attenuate slightly the inhibition of food intake induced by 10 micrograms neurotensin.     

Morphine selectively influences macronutrient intake in the rat

Dietary self-selection of the three macronutrients, protein, carbohydrate and fat, was examined in male rats following the administration of three doses of morphine sulphate: 10 mg, 15 mg, and 30 mg/kg body weight. Intakes of all three macronutrients were suppressed in a dose-dependent manner for a two-hour period following morphine administration. Both protein and carbohydrate intakes remained suppressed for a six-hour feeding period after morphine injections. In contrast, animals increased fat intake during the final four hours of the six-hour feeding period resulting in an overall increase in fat intake.     

The ontogeny of opioid receptors mediating opiate-induced feeding in rats

Acute administration of naloxone to preweanling rats does not attenuate independent ingestion of milk until 14 days of age suggesting that the full expression of an endogenous opioid system(s), regulating feeding rats, is not complete prior to this age. The present study was undertaken to examine the functional ontogeny of opioid receptors mediating opiate-induced feeding in rats. Rat pups, satiated with milk, were given intraperitoneal injections of the opiate receptor agonist, morphine, and were allowed free access to milk. Morphine stimulated the intake of milk at 3, 5, 7, 14 and 21 days of age, within 2 hr of injection. A time-course analysis in 7-day-old pups showed greater enhancement of intake between hours 2 and 4, than between hours 0 and 2, for large doses of morphine (0.3 and 1.0 mg/kg) suggesting that morphine-induced behavioral depression, which was observed early in the test session, confounded intake at earlier hours. Administration of the opiate receptor antagonist, naltrexone, produced no effect on intake of its own, but blocked the stimulation of intake by morphine in 5-day-old pups confirming that the effect of morphine on the intake of milk was mediated by opioid receptors. Thus, while a functional endogenous opioid system(s), regulating feeding in rats, is not fully mature until 14 days postpartum, the present results suggest that opioid receptors mediating feeding are functional very early in the postnatal development of the rat.     

Effect of toluene on ethanol preference in rats

The effect of toluene on the preference of ethanol was studied in rats. Toluene was administered orally by stomac tube in doses of 200, 400 or 800 mg/kg daily for 5 weeks or by inhalation at a concentration of 2000 p.p.m. 6 or 8 hr daily for 5 or 6 days per week for 2 weeks in rats of different age. During toluene inhalation exposure the rats had access to either tap water or ethanol-containing water (6 or 10%). After the exposure and during oral administration the rats had access to both ethanol-free and ethanol-containing water. Toluene inhibited the body weight gain in the highest oral dose group and in rats exposed to toluene and forced to drink ethanol in the inhalation experiments. In these experiments, the intake of fluid was reduced in the exposure period in rats forced to drink ethanol-containing water and further reduced in rats exposed to both ethanol and toluene. Exposure to toluene alone increased the fluid consumption. The preference of ethanol defined as consumed ethanol-containing water in per cent of the total water intake was not influenced by oral administration of toluene. It was, however, reduced by toluene given by inhalation to rats forced to drink ethanol-containing water during the exposure period. Toluene exposure alone or forced ethanol intake alone caused in these experiments a short-lasting reduction of the ethanol preference. It is concluded that toluene decreases the preference of ethanol in rats forced to drink ethanol during exposure to toluene.     

Characteristics of reserpine-induced suppression of NaCl solution intake in rats

Effects of single and repeated administration of reserpine on time-limited drinking of a hypertonic (1.5% w/w) NaCl solution were investigated in rats to assess whether this drug possesses anxiolytic action. Rats adapted to a 23-hr water-deprivation schedule with a free-feeding regimen were allowed a daily 1-hr water rehydration session. In the single-administration experiment, reserpine (0.1, 0.2 and 0.4 mg/kg, IP) was administered to rats at 15 min or 23 hr before a drinking session, where the fluid available was 1.5% NaCl solution. Drug was administered every 7th day. In the repeated-administration experiment, reserpine (0.1 mg/kg/day) was injected daily for 10 days 15 min before each drinking session. The fluid available was water on the first 9 days and NaCl solution on the 10th day. Reserpine suppressed NaCl solution intake when it was singly administered at 15 min before the rehydration, whereas no significant change in the fluid intake occurred when it was administered 23 hr before drinking, even though rats showed ptosis in response to 0.2 and 0.4 mg/kg doses. Tolerance developed to the suppressing effect of repeated administration of reserpine on fluid intake, although ptosis and sedation continued and body weights decreased. Tolerance was almost complete after 11 days. The results suggest that reserpine does not have an anxiolytic effect.     

A shift to diurnal feeding following chronic administration of theophylline in rats]

Influence of chronic administration of theophylline on feeding behavior in rats was studied in contrast with the saline controls under a 12 hr light-dark cycle. The results obtained were as follows: 1) total amount of food intake per 24 hr was not affected by either chronic administration of theophylline or saline administration; 2) theophyllinized rats, however, ate more food during light time (0900-2100) and less food during the dark (2100-0900); 3) the amount of food intake during the dark in the control group was approximately 80% of total intake, while that in the theophylline group decreased to below 60%; 4) increase in ratios of body weights in the theophylline and the control groups did not differ significantly throughout the entire experimental period. Feeding behavior of the rat is closely correlated with a daily light-dark cycle as was seen in the control group. The feeding circadian pattern disappeared in theophyllinized rats, i.e., the behavioral characteristic of a nocturnal feeder could not be preserved following theophylline administration. The disappearance of the feeding cycle may result from a phase shift of the cycle, which was induced by facilitatory effects of theophylline on food intake during light time.     

Effects of methylphenidate on food and water consumption at different body weights.

The effects of intraperitoneally administered methylphenidate at 0, 1.5, 3.0, 6.0, and 12.0 mg/kg were studied in two experiments. Experiment 1 determined the effects of methylphenidate on 0.5, 1.0, 2.0, and 24 hr post injection food and water consumption in rats at ad lib feeding body weights. Experiment 2 determined the post injection effects of methylphenidate on 0.5, 1.0 and 2.0 hr food and water and 24 hr water consumption in rats maintained at 80% ad lib feeding body weight due to partial food deprivation. The results of Experiment 1 indicate that when animals are feeding ad lib at normal body weight food and water consumption is decreased for 2 hr following the administration of the lowest 1.5 mg/kg dose of methylphenidate. Methylphenidate in doses as high as 12.0 mg/kg has no effect on 24 hr food and water consumption under these conditions. The results of Experiment 2 indicate that when animals are maintained at reduced body weight due to partial food deprivation, food consumption for 2 hr is significantly decreased by the highest, 12.0 mg/kg, dose of methylphenidate. These effects are observed within the first 30 min post injection when methylphenidate decreases food consumption in a dose dependent manner. Methylphenidate has no effect on water consumption under these conditions. The effects of methylphenidate on ingestive behavior are discussed in terms of previous experiments and the possible differential effects on motor activity at different body weights under different stimulus conditions.     

Differential effects of morphine on food and water intake in food deprived and freely-feeding rats

In two experiments the effects of a range of doses of morphine (1, 3, 10 and 30 mg/kg) on the food and water consumption of rats were studied. The results of the first experiment showed that in 24 h food-deprived rats, morphine reduced levels of food and water intake. The duration of these actions was dependent upon dose, with only the highest dose (30 mg/kg) producing any effect persisting for longer than 4 h. In contrast a second experiment showed that morphine increased levels of food and water intake in non-deprived animals. The effect on food intake was most apparent when measurements were taken at 2 h and 4 h after drug administration, while water intake remained above control levels for over 6 h. This study shows that the actions of morphine on ingestion of food and water are affected by food deprivation, and the results are consistent with the hypothesised role of endogenous opiates in the mediation of such behaviour.     

Physical dependence on morphine, phenobarbital and diazepam in rats by drug-admixed food ingestion.

To produce physical dependence on morphine, phenobarbital and diazepam in rats, these drugs were mixed with powder form of rat food in concentrations of 0.5 mg/g, 1 mg/g and 2 mg/g of food. One group of rats (the lower dose group) was continuously exposed for 1 week to two morphine-admixed foods with morphine to food ratios of 0.5 mg/g and 1 mg/g in a cage. The other group (the higher dose group) could choose between two morphine-admixed foods with morphine to food ratios of 1 mg/g and 2 mg/g. After 1 week, morphine-admixed foods were replaced with morphine free food for 2 days. Both groups of rats showed greatly reduced body weight and food intake after the first 24-48 hr withdrawal. The body weight decrease was greater for rats in the higher dose group. Control groups of morphine dependent rats were kept on the morphine added food diets and showed the same body weight increase as well as normal control rats during the course of these experiments. Physical dependence on phenobarbital and diazepam was produced using the same dosage schedules as with morphine. Both the lower and higher dose groups showed significant decrease in body weight due to withdrawal after 1 week of drug-food exposure. Levallorphan (0.5, 1, 3 and 5 mg/kg, s.c.) administered to morphine dependent rats had dose-dependent effects on the intensity of abstinence symptoms (e.g., diarrhea, piloerection and wet shakes phenomena), maximal decrease in body weight and duration of decreased body weight. Cross-physical dependence between phenobarbital and diazepam was demonstrated by this method.     

Morphine-stimulated feeding: analysis of macronutrient selection and paraventricular nucleus lesions

The hypothalamic paraventricular nucleus (PVN) has been found to be sensitive to the feeding stimulatory effects of opiates. The present experiments investigated the effect of systemic morphine (2 mg/kg) on macronutrient selection in freely-feeding and food-restricted rats and assessed the impact of PVN electrolytic and 6-hydroxydopamine lesions on the rats' ability to respond to peripheral morphine injection. In satiated rats, maintained ad lib on pure macronutrient diets, morphine increased food intake. This effect was associated with a preferential increase in protein ingestion; carbohydrate consumption, compared with fat and protein intake, was least affected. In food-restricted rats, permitted to eat for 6 hr, morphine instead produced a particular preference for fat, with no significant enhancement of total calorie intake. While PVN 6-hydroxydopamine lesions, which depleted PVN catecholamine levels by 70%, failed to alter morphine-stimulated feeding, electrolytic lesions of the PVN significantly attenuated this response, particularly protein and fat ingestion. This suggests that opiate-induced feeding may, in part, be mediated through the PVN, which is known to have an important function in the control of food ingestion.     

Circulating naphthionic acid in nonpregnant and pregnant rats after feeding amaranth

Naphthionic acid (NA) [1-naphthylamine-4-sulfonic acid], is one of the azoreduction products absorbed from the rat's gut after oral ingestion of amaranth [FD&C Red No. 2, C.I. 16185; CAS [915-67-3]]. This report describes the blood levels of NA in male and nonpregnant female rats as well as the level of NA in fetal blood, amniotic fluid, and maternal blood of rats fed various doses of amaranth. Three groups of five male and eight female rats were fed diets for 9 days to provide estimated daily doses of 20, 200, and 2000 mg amaranth/kg body wt. Five male and six female rats were fed a control diet. Blood samples were collected twice daily after 1, 2, 3, 4, 5, and 8 days of feeding and analyzed for NA. After the ninth day, the male rats were fasted and blood was collected at specific intervals over the next 96 hr. The female rats were mated with untreated males and received the same dosages of amaranth throughout mating and gestation. The amniotic fluids and blood from each fetus as well as the maternal blood were collected between the 19th and 21st day of gestation for the NA determination. A significant elevation in plasma NA concentrations occurred in male and female rats receiving estimated doses of 200 or 2000 mg amaranth/kg body wt. Plasma NA concentrations were slightly but not significantly increased at 20 mg amaranth/kg body wt. The NA concentrations in fetal plasma and amniotic fluid from the 2000 mg amaranth/kg body wt dose group were significantly less than (p < 0.05) 10-fold higher than those from the 200 mg amaranth/kg body wt dose group. The NA concentrations in maternal blood from the 2000 mg amaranth/kg body wt dose group were less than (although not significant statistically) 10-fold higher than from the 200 mg amaranth/kg body wt dose group. The NA concentrations in maternal plasma were about five times higher than in plasma of their fetuses. There was a significant increase in the NA concentrations in amniotic fluid between the 20th and 21st day of gestation.