Role of behavioral and pharmacological variables in the loss of tolerance to amphetamine hypophagia

 Tolerance to amphetamine-induced hypophagia is lost when drug injections are withdrawn for 4 weeks while milk tests are continued (Wolgin and Hughes 1996). The purpose of this study was to determine whether the loss of tolerance is a function of drug withdrawal per se. Rats made tolerant to amphetamine (2 mg/kg, IP) were assigned to one of three groups. During the next 4 weeks (phase), one group continued to receive amphetamine injections prior to daily milk tests (Before group), one group received drug injections after the milk tests (After group), and one group received injections of saline prior to the milk tests (Saline group). Dose-response tests revealed that the Before group retained tolerance, whereas the After and Saline groups lost tolerance. When retested with chronic injections of amphetamine prior to milk, the After and Saline groups reacquired tolerance more rapidly, and to a greater extent, than non-tolerant controls. These results demonstrate that the loss of tolerance is not due to drug withdrawal per se, but may be due to the unlearning of behavioral strategies previously acquired under the drug. Received: 17 September 1996 / Final version: 10 March 1997     

Sensitization to haloperidol-induced suppression of milk intake: effect of interdose interval

The purpose of this study was to determine the effect of manipulating the interdose interval (IDI) on the suppression of milk intake induced by haloperidol (HAL). Groups of rats were given chronic injections of either HAL (0.625 mg/kg) or saline at IDIs of 1, 2, 7, or 14 days. Dose-response curves were determined at the conclusion of the chronic phase. The results indicated that injections of HAL given at IDIs of 1 or 2 days produced neither tolerance nor sensitization, whereas injections given at intervals of 7 or 14 days produced sensitization. Sensitization was also observed in the control groups, perhaps as a result of the intermittent schedule of HAL injections given during the dose-response tests. Sensitization to HAL was not accompanied by changes in sensitivity to amphetamine. The results of this experiment are consistent with those of other studies in showing that the behavioral effects of neuroleptics are strongly influenced by the schedule of injections. In addition, evidence is presented that sensitization to HAL-induced hypophagia is contingent on behavioral experience under the drug.     

Role of associative and nonassociative mechanisms in tolerance to morphine "anorexia"

To determine whether tolerance to morphine-induced anorexia involves associative mechanisms, rats were given chronic injections of morphine (Group 1, 10 mg/kg; Group 2, 20 mg/kg) in the presence of one compound cue on alternate days and injections of saline in the presence of another compound cue on the intervening days. After tolerance developed to the initial suppression of intake, three tests of Pavlovian conditioning were conducted. On the compensatory response test, in which saline injections were given in the presence of the morphine cue, only Group 2 showed a significant increase in milk intake. On the explicit unpairing test and the environmental specificity test, in which morphine injections were given in the presence of the saline cue or in an entirely different room, respectively, neither group showed a significant loss of tolerance. The failure to demonstrate cue-dependent tolerance in this paradigm may have been due in part to inadvertent temporal conditioning and in part to the rapid development of nonassociative tolerance.     

Tolerance,withdrawal, and supersensitivity to dopamine mediated cues in a drug-drug discrimination

Rats were trained to discriminate between 0.25 mg/kg amphetamine (AMPH) and 0.03 mg/kg haloperidol (HAL) in a two-lever drug discrimination task. In order to test for a drug-induced withdrawal state, animals were assigned to one of three chronic treatment groups and given injections of AMPH, HAL, or distilled water (DW) for 10 consecutive days. Subjects from each treatment condition were then tested at 24, 48, or 72 h after the final injection. At the 24 h retest interval, subjects injected with AMPH responded as though administered an acute dose of HAL (0.028 mg/kg) and subjects injected with chronic HAL responded as though administered an acute dose of AMPH (0.15 mg/kg). By 72 h choice behavior had returned to pretreatment values. To determine whether the rebound observed after 10 days of drug treatment was present after a single injection, independent groups of subjects were injected with single doses of either 10 mg/kg AMPH or 1.0 mg/kg HAL and then retested from 4 h to 48 h later. Single doses of both AMPH and HAL produced significant rebounds that peaked between 20 h (AMPH) and 24 h (HAL) following administration. In a third experiment, animals were tested with or without acute doses of drug following pretreatment with either HAL or AMPH. Receptor supersensitivity accounts for the tolerance observed to HAL 24 h after treatment with 1.0 mg/kg HAL, whereas receptor subsensitivity accounts for the tolerance observed 20 h after treatment with 10 mg/kg AMPH.Some of the data presented here was presented at the meeting of the Society for Neurosciences, New Orleans, 1991.     

Development and reversal of sensitization to amphetamine-induced hypophagia: role of temporal,pharmacological, and behavioral variables

This study shows that sensitization can develop to amphetamine-induced hypophagia and examines the stability of this effect following subsequent pharmacological and behavioral experience. Rats given 36 injections of either amphetamine (2.5 mg/kg; Group A) or saline (Group S) at 3-day intervals developed sensitization of hypophagia, as assessed by a shift to the left in the dose-response (DR) function. Group A also displayed sensitization of stereotypy, whereas Group S showed little change except at the highest dose. Subgroups from each group were then given daily injections of amphetamine (2 mg/kg) either before or after access to milk for 4 weeks. Other subgroups were given injections of saline as a control. On a final DR determination, these control groups showed no further changes in milk intake. In contrast, groups given chronic injections of amphetamineafter milk showed a loss of sensitization (DR3=DR1), whereas groups given the drugbefore milk developed tolerance that was limited to the chronic dose. These results demonstrate that (1) sensitization of amphetamine-induced hypophagia and stereotypy can develop independently; (2) sensitization of hypophagia can be reversed, without inducing tolerance, by subsequent daily exposure to the drug; and (3) prior sensitization of hypophagia does not preclude the subsequent development of tolerance if the drug is later given in the context of feeding.     

Effects of repeated administration of serotonergic agonists on diet selection and body weight in rats

Food intake, diet selection and body weight gain were examined in three separate experiments in which rats received saline or one of three serotonergic agonists, dexfenfluramine, RU 24969 and fluoxetine. In all experiments, food was available only in the dark period during which time rats were given simultaneous access to two isoenergetic diets which differed in their protein and carbohydrate content. After habituation to this feeding paradigm and intraperitoneal injections, rats were assigned to control or drug group. Saline or a serotonergic agonist was given to the same rat once daily, 15 min prior to feeding, for six consecutive days. All three agonists (1.5 mg/kg for dexfenfluramine and RU 24969; 3 mg/kg for fluoxetine) caused immediate (first two h of feeding) hypophagia which was accounted for by the selective suppression in intake of the high-carbohydrate-low-protein diet. This selective shift in diet choice was sustained upon repeated exposure. Although the effects of these agonists on daily (12-h) feeding was less pronounced, appetite suppression was due entirely to reduced intake of the high-carbohydrate-low-protein diet. Of the three agonists tested, partial tolerance was observed only after dexfenfluramine. Nevertheless, all three agonists caused comparable declines in weight gain. These results suggest that repeated administration of serotonergic agonists has sustained impacts on food intake, diet choice and weight gain.     

Sensitization of haloperidol-induced hypophagia is contingent on behavioral experience with food

Groups of rats were given injections of haloperiodol (0.31mg/kg) at weekly intervals either before or after access to sweetened milk. Control groups were given injections of saline. At the end of the chronic regimen, all groups received a single injection of haloperidol (0.15mg/kg) prior to milk access. Rats injected with the drug before milk during the chronic phase showed a progressive decrease in milk intake. When subsequently challenged with a lower dose, this group ingested less milk than any of the other groups, which did not differ from one other. These results demonstrate that sensitization of haloperidol-induced hypophagia is contingent on experience with milk while in the drugged state.     

Role of dose interval in the acquisition of tolerance to methylphenidate

Two experiments were performed to test the role of dose interval in the development of tolerance to methylphenidate. Rats were trained to consume sweetened milk and then were given methylphenidate in a dose that decreased milk intake by approx. 50%. For the next 23 sessions they received either saline; methylphenidate daily, immediately post-session; or pre-session methylphenidate, either daily, every-other-day, or every-four-days. The next session, all groups received methylphenidate pre-session. The 3 groups treated on a chronic basis with methylphenidate pre-session returned to baseline levels of milk intake and differed significantly from the daily saline and daily post-session methylphenidate groups, which did not become tolerant. In a second experiment, rats injected presession daily or every fourth day with 15mg/kg methylphenidate developed tolerance to the disruption of milk consumption. As compared to rats treated chronically with saline, the 2 groups given methylphenidate showed a shift of their dose-effect curves to the right and cross-tolerance to d-amphetamine. These results demonstrate that tolerance can occur to the disruptive effects of amphetamine-type drugs even when three drug-free days intervene between administrations. This tolerance is characterized by a shift in the dose-effect curve as well as cross-tolerance to a drug with similar pharmacological properties.     

Changes in behavioural contingencies produce a loss of tolerance to amphetamine hypophagia in rats despite continued feeding tests while drugged

Rats administered amphetamine prior to access to milk in bottles develop tolerance to the hypophagic effect of the drug by learning to suppress stereotyped behaviours that interfere with feeding. When tolerant rats are later allowed to drink milk from a bottle in an unintoxicated state, tolerance is lost, even when drug exposure is held constant by administration of the drug after the test. In the present experiment, we show that tolerance can also be lost in the face of continued administration of amphetamine prior to milk tests, as a result of changes in the contingencies of reinforcement that govern the suppression of stereotypy. Rats were injected with 2 mg/kg amphetamine and given access to milk in bottles for 16 trials. Tolerance to the hypophagic effect was confirmed by dose-response tests in which milk was available in bottles. The rats were then injected with 2 mg/kg amphetamine prior to intraoral milk infusions for 21 trials. This method of feeding did not require the suppression of stereotypy to obtain milk. Subsequent dose-response tests in which milk was again presented in bottles revealed that tolerance was lost, even though intoxicated feedings were never interrupted. These results demonstrate that the contingencies of reinforcement governing the suppression of stereotypy determine whether tolerance is retained or lost.     

Behavioral tolerance to flurazepam

Rats were trained to earn 180 food pellets in daily, fixed-interval 1-min sessions. When performance had stabilized, a Before group was given a weekly 16 mg/kg flurazepam injection IP for 3 weeks immediately before the sessions, while an After group received their weekly injections immediately after the sessions. Then, the After group received 3 such weekly injections before the sessions. Behavioral tolerance developed by the 2nd flurazepam injection for the Before group, but for the After group, the 3 postsession flurazepam injections resulted in subsequent tolerance to presession flurazepam injection for session lever presses, but not for the time taken to earn 180 pellets. Dispositional tolerance to the serum elimination rate of flurazepam did not develop over the course of 3 injections. Behavioral suppression still evident in the initial portion of sessions with the 2nd and 3rd presession injection coincided with the duration of rising and high levels of serum flurazepam.     

Long-term retention of tolerance to amphetamine hypophagia following cessation of drug injections and feeding tests.

According to the instrumental learning model, tolerance to amphetamine hypophagia involves learning to suppress stereotyped movements that interfere with feeding. If both drug injections and feeding tests are then suspended, learning should be retained and no loss of tolerance should occur. However, previous studies have only assessed the retention of tolerance for 3-4 weeks. In the present study, retention intervals of 4-31 weeks were used. Rats were given daily injections of amphetamine (2 mg/kg) and access to milk for 30 min until tolerance developed to drug-induced hypophagia. Yoked controls were injected with saline. Both before and after this phase, dose-response (DR) tests were conducted. Drug injections and feeding tests were then suspended. At 4, 10, 18, and 31 weeks, both groups were injected with 2 mg/kg amphetamine and given access to milk for 30 min to assess the retention of tolerance. A final DR determination was then conducted. Most (88%) rats retained tolerance to 2 mg/kg amphetamine for 31 weeks. However, DR tests revealed that tolerance was not retained at 4 mg/kg. The results demonstrate that learned tolerance to amphetamine can be retained over long intervals when both drug injections and feeding tests are suspended.     

Effects of chronic haloperidol on stress-induced oral behaviour in rats

Rats received daily injections of haloperidol (HAL, 5 mg/kg SC, or IP) or tartaric acid vehicle for 14–21 days. Four to six days after discontinuation of the daily injections, rats were given a single 5-min tail pinch (TP) test. HAL-treated rats showed significantly shorter latencies to display oral behaviours (OB: licking, gnawing, or drinking) compared to controls in five separate replications. Food consumption per se was not consistently affected. Shorter OB latencies were significantly corrlated both with increased striatal ³H-spiperone binding and with apomorphine stereotypy scores. In a final eperiment, this effect of HAL on OB latencies was blocked by a systemic injection of naloxone (2 mg/kg IP) prior to the TP test. Naloxone did not affect OB latency in control rats, suggesting the possibility of endogenous opiate involvement in the chronic HAL effects. The overall results suggest that OB latencies following mildly activating stimulation may provide a useful behavioural assay for neuroleptic-induced oral abnormalities as well as a functional index of striatal dopamine D-2 receptor sensitivity.     

Loss of tolerance to amphetamine-induced hypophagia in rats: homeostatic readjustment vs. instrumental learning.

According to the homeostatic model, the loss of tolerance to amphetamine-induced hypophagia requires a period of unrestricted feeding in the drug-free state, which transforms the compensatory response mediating tolerance ("hyperhunger") into a functional disturbance to homeostasis. In the absence of such a disturbance, tolerance should be retained. To test this prediction, rats tolerant to amphetamine's hypophagic effect were given a 4-week tolerance retention period during which milk intakes were restricted and deprivation levels held relatively constant. During this period the rats were assigned to one of the following drug treatment conditions: 1) saline injections both before and after daily milk tests (saline group); 2) saline injections before, and amphetamine injections after, daily milk tests (after group); 3) no injections and no milk tests (no-treatment group); or 4) amphetamine injections before, and saline injections after, milk tests (before group). Despite the restricted feeding regimen, both the saline and after groups lost tolerance. These results do not support the homeostatic model, but are consistent with the instrumental learning model, which views drinking milk in the undrugged state as analogous to receiving noncontingent reinforcement.     

Tolerance to the discriminative stimulus properties of d-amphetamine

Male rats (F-344) responding for milk on a VI 20 sec schedule of reinforcement were trained to discriminate which of two levers to press on the basis of whether they had been injected with d-amphetamine (0.50, 1.00 or 1.50 mg/kg) or saline 15 min prior to daily training sessions. Dose-response functions determined for each of the three (n = 6) training-dose groups indicated that ED₅₀ values were directly correlated with training dose. Two days following chronic amphetamine injections (a total of 78 mg/kg over 4 days) rats were tested for tolerance at a dose which normally produced about 80% drug-lever responding. Rats in all three groups showed tolerance to the cue properties of amphetamine. In the 0.50 and 1.00 mg/kg groups, complete tolerance was shown as evidenced by the fact that the drug lever responding did not differ from that which was appropriate following saline injections.     

Differential tolerance development to buprenorphine-, Diprenorphine-, and heroin-induced disruption of food-maintained responding in Macaque monkeys

Single daily subcutaneous injections of buprenorphine (1.0 mg/kg), diprenorphine (1.0 mg/kg), or heroin (1.0 mg/kg) were given over 25 consecutive days to examine the degree and the rate of tolerance development to drug-induced suppression of food maintained responding. One gram food pellets were available on a second order schedule (FR 4 VR 16:S) during the 1-hr sessions three times a day. All drug and saline control injections were given at 10:00 a.m., 1 hr before a food session. During the first three days of treatment all three drugs produced marked suppression of food-maintained performance. Recovery from buprenorphine-and diprenorphine-induced suppression of food-maintained responding occurred within four and eight days, respectively. By the 25th day of buprenorphine and diprenorphine treatment, operant responding for food increased significantly above control levels (p<0.01). In contrast, the significant heroin-induced disruption of food-maintained responding (p<0.01) persisted throughout the 25-day treatment period. Saline substitution for all three drugs resulted in a gradual return to control levels of food pellets earned. Linear regression analysis of the linear portion of the time-effect curve revealed significant differences in both the rate and the degree of tolerance development of these three drugs. These differences in tolerance development may reflect pharmacokinetic differences between the relatively short-acting heroin and the longer-acting diprenorphine and buprenorphine.     

Naloxone and fluid consumption in rats: dose-response relationships for 15 days

Rats were given daily intraperitoneal injections of 10.0, 1.0, 0.1, 0.01, 0.001 or 0.0 mg/kg naloxone for 15 days. Each day after the injections, animals were allowed access to a 20% sucrose solution for two hours and to tap water for the subsequent 10 hours. Consumption of the sucrose solution by the group that received 1.0 mg/kg was reliably decreased on Day 1 and 2, reflecting the suppressive effect of naloxone at that dose. By Day 3 until the end of the experiment, however, the suppression was no longer significant, suggesting that tolerance had developed. A similar effect was seen with the group given the highest dose, 10.0 mg/kg; although drinking was significantly less than the control in each of the 15 sessions, this group showed a trend to increase intake over the days of the experiment, thus also indicating possible tolerance to the effect of naloxone. Drinking patterns of the other groups did not differ statistically from the control. Thus, the low doses had no ability to suppress consumption, and the lowest dose that did lower it soon lost that ability; the highest dose continued to suppress drinking throughout the study but with decreasing efficacy. High performance liquid chromatography (HPLC) demonstrated that the naloxone remained intact over the 15 days of the experiment, supporting the suggestion that tolerance to naloxone might have developed.     

Tolerance and cross-tolerance to the effects of amphetamine, methamphetamine and fenfluramine on milk consumption in the rat.

It has been reported that rats develop tolerance to the milk intake suppressant effects of d-amphetamine, d-methamphetamine, and d, l-fenfluramine. However, it has been hypothesized that the mechanism of action of fenfluramine is different from that of the other two drugs. In the present experiment, rats were given one of these 3 phenylethylamine derivatives each day until tolerance developed to the suppression of milk intake. A second of these drug s was then substituted and milk intake measured. There was cross-tolerance to the drug-induced suppression between d-amphetamine and d-methamphetamine, but not between d-amphetamine and d,l-fenfluramine. The lack of cross tolerance suggests a different mechanism of action for these drugs.     

Effects of acute and chronic cocaine on milk intake, body weight, and activity in bottle- and cannula-fed rats

The effects of cocaine on the milk intake, body weight and activity of bottle- and cannula-fed rats was compared under both acute and chronic dosing conditions. Bottle-fed rats were initially more hypophagic than cannula-fed rats when given acute injections of cocaine (4-40mg/kg). Following chronic injections of the drug (16mg/kg), bottle-fed rats developed tolerance, as shown by a rightward shift in the dose-response function for milk intake. Such tolerance was accompanied by a decrease in drug-induced motor activity. In contrast, cannula-fed rats showed marked sensitization of stereotyped movements. Bottle -fed rats showed marked sensitization of stereotyped movements. However, weight loss per se was not a determining factor in tolerance development, because cannula-fed rats given chronic injections of 32mg/kg cocaine lost even more weight, but did not become tolerant. These results suggest that, at moderate doses, cocaine suppresses feeding primarily by inducing behaviors that are incompatible with the appetitive phase of feeding, and that tolerance involves learning to inhibit such responses in order to feed.     

Effects of variation in chronic dose of cocaine on contingent tolerance as assessed in a milk-drinking task

Tolerance to the suppressive effects of cocaine on milk drinking by rats was studied using a contingent tolerance experimental design. Three separate groups (n=6) of rats received 8.0, 16.0, or 32.0 mg/kg cocaine daily 15 min before a 15-min period of access to sweetened condensed milk for 20 days. Three additional groups of six rats each received the same chronic doses 15 min after access to milk. Milk, water, and food intake as well as body weight were measured daily. Tolerance effects were assessed by comparing initial acute dose-effect determinations with a probe dose-effect redetermination in which all rats again received doses of cocaine pre-session after having experienced the differential pre- or post-session chronic treatment. Behavioral tolerance on the milk intake measure was observed for the 8.0 mg/kg and 16.0 mg/kg doses, but not for the 32.0 mg/kg chronic treatment, even though the latter group exhibited evidence of tolerance in the water intake measure. Chronic treatment with 8.0 and 16.0 mg/kg produced different outcomes in that chronic exposure to 16.0 mg/kg in the presence of milk resulted in generalization of tolerance to both a lower (8.0 mg/kg) and a higher dose (32.0 mg/kg), but the group receiving 8.0 mg/kg did not exhibit generalization of tolerance to higher doses. Modest sensitization effects were observed in the rats treated post-session with either 8.0 or 16.0 mg/kg. Rats receiving 32.0 mg/kg pre-milk exhibited a dramatic lowering of food intake and concomitant loss in body weight compared to the same dose given after the milk access period, suggesting that pairing cocaine with a highly palatable substance may enhance its toxic effects. The observed behavioral tolerance effects were discussed in terms of (1) the reinforcement loss hypothesis, (2) the channeling hypothesis developed in the context of behavioral tolerance to amphetamine, and (3) the concept of dose-specific tolerance.Supported by DA 05/253     

Time course for the induction and maintenance of tolerance to Delta(9)-tetrahydrocannabinol in mice

The time course for the development of tolerance to delta-9-tetrahydrocannabinol (Delta(9)-THC) was studied in an effort to determine the role that length of dosing may have in the onset and maintenance of tolerance. Mice were chronically treated with either vehicle or 10 mg/kg of Delta(9)-THC subcutaneously twice a day. The mice were tested 24 h after the last injection for tolerance as assessed by the production of antinociception and suppression of spontaneous activity. Tolerance was first observed after three injections of Delta(9)-THC (1.5 days) resulting in a 7-fold and 23-fold decrease in potency for the measures of antinociception and hypoactivity, respectively. Seven injections of Delta(9)-THC (3.5 days of dosing) resulted in a 12-fold and 36-fold decrease in potency, respectively, while 13 injections of Delta(9)-THC (6.5 days of dosing) produced a 6.2-fold and 9.8-fold degree of tolerance. The time course for the recovery from Delta(9)-THC-induced tolerance was also determined with a separate group of animals. Mice were dosed for 6.5 days with 10 mg/kg of Delta(9)-THC and were not tested until 4.5, 7.5, and 11.5 days after cessation of drug treatment. After 4.5 days without drug treatment the mice exhibited a 7.5-fold and 2.3-fold degree of tolerance as measured by antinociception and hypoactivity, respectively. After 7. 5 days without drug treatment a 3.4-fold degree of tolerance remained for the measure of antinociception, while no tolerance was detected for the measure of hypoactivity. No tolerance was observed for the measure of antinociception after 11.5 days without drug treatment. This time course indicates that the mechanisms responsible for either the production or maintenance of tolerance differ between the measures of antinociception and suppression of spontaneous activity.