An analysis of the effects of contextual cues on the development of morphine tolerance in rats.

Tolerance to morphine analgesia was determined daily by exposing rats either to the same box or to different boxes by a repeated administration of morphine (5 mg/kg). In the Acquisition Phase, the rats received either morphine or saline in the same or different boxes for four consecutive days, and the process of tolerance development was assessed by a hot-plate test. Marked tolerance developed in the group exposed to the same contextual cue, whereas tolerance was attenuated in the group exposed to different cues. In the Extinction Phase, all rats received saline injections in the box exposed on Day 1 for four days. On the first day, hyperalgesia was observed only in the rats injected with morphine in the same contextual cue. In the Retest Phase, the rats underwent a second morphine injection and to some extent showed recovery from tolerance. In the Acquisition Phase, the number of animals showing abnormal activity with morphine injection increased monotonically in the group that was administered morphine in the same box (Group M-S) before injection, but in the group administered the drug in different boxes (Group M-D), no systematic development of the activity occurred. These results indicate that the hindering of a rat's ability to associate with environmental cues under the effect of morphine slows the development of tolerance, and the withdrawal and anticipatory symptoms, and the tolerance of morphine involves psychological and pharmacological factors.     

Sensitivity to the narcotic cue in non-dependent, morphine-dependent and post-dependent rats

Using a food-reinforced two-lever operant procedure, 12 rats were trained to discriminate 10 mg/kg (i.p.) of morphine from saline. Five animals were given daily non-contingent exposure to morphine (20 mg/kg on saline, or no-test days, and 10 mg/kg on drug days) from the beginning of the experiment; the others received injections of saline. In the morphine generalization tests, the dependent rats showed an increased sensitivity to the narcotic cue as compared with non-dependent animals (ratio of the ED50 values: 2.30). This increased sensitivity was still present 3 months after discontinuing the non-contingent treatment with morphine (ratio of the ED50 values: 1.98). The results of the present study, together with other results reported in the literature, suggest that the experimental procedure plays a role in determining whether tolerance, no tolerance or enhanced sensitivity to the discriminative stimulus properties of narcotics, is observed.     

Role of drug-administration cues in the associative control of morphine tolerance in the rat

This research investigated the role of injection procedures as a potential confound in the study of associative and nonassociative morphine tolerance development. Rats administered a series of morphine injections paired with a distinctive context environment can develop tolerance controlled associatively by the context. However, rats given morphine unpaired with the context may also develop some degree of tolerance. This study examined whether this tolerance represents an associative effect with animals using the injection ritual as a cue predictive of morphine delivery. Following 14 days of habituation to handling and injection stimuli, rats were given eight morphine injections (20 mg/kg, IP) explicitly paired or unpaired with a distinctive context. Animals were then tested for morphine analgesia in the context after either a 30-day rest condition or a 30-day period of daily saline injections. Analgesia was assessed by the tail-flick method, and tolerance was defined as the shift to the right of the dose-response curve of morphine-experienced relative to saline control animals. Paired animals across both retention conditions displayed tolerance, whereas tolerance retention in unpaired animals was observed only in those animals not given saline injections over the 30-day interval. Results support an associative interpretation of tolerance observed in unpaired conditions and suggest that the injection ritual may provide highly salient cues for the support of associative tolerance effects.     

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.     

Magnitude of initial effect influences development of tolerance to morphine in rats responding under a fixed-ratio schedule of food presentation

This experiment examined whether development of tolerance to the rate-decreasing effects of morphine can be modulated by the magnitude of the initial effect of morphine. Lever pressing by rats was maintained under a fixed-ratio 30 schedule of food delivery in daily 30 min sessions. Subjects were assigned to two groups, which did not differ in initial sensitivity to morphine, and given daily injections of morphine under dosing schedules chosen to have different initial effects. Group 1 received daily pre-session injections of 10 mg/kg morphine, which initially suppressed lever pressing completely, and post-session injections of saline. Group 2 received morphine in two injections, one pre-session and one post-session. The initial daily doses 10 mg/kg pre- and 9 mg/kg post-session) decreased rates by 30% or less. The proportion of the 10 mg/kg dose administered pre-session was increased gradually over 10 weeks. Group 2 developed greater tolerance than did group 1, as assessed by changes in the dose of morphine required to suppress response rates by > 50%. After 3 months of repeated treatment, the ED(50) of morphine increased 2.5-fold in group 2, but only 1.4-fold in group 1. When the daily dose of morphine was raised to 20 mg/kg (10 mg/kg pre- and 10 mg/kg 14 h post-session) for all subjects, the ED(50) of morphine increased 6.7-fold in group 2, but only 2.6-fold in group 1. During repeated treatment, the groups did not differ in the dose of maloxene required to suppress response rates or elicit weight loss. Following termination of morphine treatment, the ED(50) of morphine returned to original values in group 2, but was 1.8-fold lower than initial values in group 1. Thus, a pronounced [??103] decreasing effect of morphine retarded development of tolerance, perhaps by a conditioning process.     

Effects of Pavlovian conditioning and MIF-I on the development of morphine tolerance in rats

Thirty male Sprague-Dawley-derived rats were given daily IP injections of morphine (5.0 mg/kg) in the presence of a specific set of environmental cues for eleven consecutive days. Twelve hours after each morphine session, a control injection was given in a different environment. On Day 12 through 14 the environmental cues associated with each session were reversed. On Day 15 environmental cues associated with each session were the same as on Days 1-11. Analgesia was assessed by the tail-flick method 30 minutes after each morphine and control injection. Four independent groups (n=6) received either a lower (0.1 mg/kg) or a higher (5.0 mg/kg) dose of MIF-I either 10 minutes before or immediately after each morphine and control session. A control group received an injection of a diluent vehicle both before and after each session. None of these peptide-treatments significantly affected either acute action of morphine or the development of tolerance across days. Tail-flick latencies from both morphine and control sessions significantly decreased across days. On Day 12, when morphine was administered in the presence of cues not previously associated with its administration, tail-flick latencies were significantly longer than on the previous day. Tail-flick latencies did not change from Day 11 to Day 15 during control sessions. Morphine-session latencies did not change from Day 14 to Day 15, although they did decrease from Day 12 to Day 14. The significant morphine-induced analgesia on Day 15 of the experiment increases a remarkable resistance to the development of tolerance to morphine. The results partially support the hypothesis proposed by Siegel [115-18] that principles of Pavlovian conditioning exert an important influence on the development of tolerance to morphine.     

Anxiolytic-like effects of morphine and buprenorphine in the rat model of fear-potentiated startle: tolerance,cross-tolerance,and blockade by naloxone

Rationale Morphine and buprenorphine have analgesic and anxiolytic-like properties. While their analgesic effects have been well characterized, their anxiolytic-like properties have not. Objectives Effects of acute morphine and buprenorphine on the expression of acoustic fear-potentiated startle (FPS) and naloxone pretreatment were assessed. Effects of chronic morphine and buprenorphine on tolerance, cross-tolerance, and withdrawal were also examined. Materials and methods Fear-conditioned rats were given subcutaneous drug treatment immediately before testing for FPS. Experiment 1, rats were administered morphine (0.03, 0.25, 0.63, 2.5, or 10 mg/kg) or buprenorphine (0.004, 0.0075, 0.015, 0.03, or 0.25 mg/kg). Experiment 2, rats were given saline or naloxone (0.5 mg/kg) and 5min later given saline, morphine (2.5 mg/kg), or buprenorphine (0.03 mg/kg). Experiment 3, rats received once-daily injections of saline, morphine (10 mg/kg), or buprenorphine (0.25 mg/kg) for 7 days. Immediately before testing, saline-treated rats were given saline, morphine (2.5 mg/kg), or buprenorphine (0.03 mg/kg), morphine-treated rats were given morphine (2.5 mg/kg) or buprenorphine (0.03 mg/kg), and buprenorphine-treated rats were given buprenorphine (0.03 mg/kg) or morphine (2.5 mg/kg). Tolerance and cross-tolerance in analgesia were assessed via the tail-flick test, as were naloxone-precipitated withdrawal. Results Morphine and buprenorphine had parallel dose–response curves in blocking FPS, with buprenorphine 40 times more potent than morphine. Naloxone reversed these effects. Morphine and buprenorphine showed tolerance and cross-tolerance in their anxiolytic-like and analgesic effects. Chronic buprenorphine produced less withdrawal than chronic morphine. Conclusions Cross-tolerance between morphine and buprenorphine suggests a common receptor mediating their anxiolytic-like and analgesic effects.     

Early neonatal experience of Long-Evans rats results in long-lasting changes in reactivity to a novel environment and morphine-induced sensitization and tolerance.

In Long-Evans rats, daily 3-h separation from the dam during the neonatal period results in enduring alterations in behavioral and neuroendocrine responses to stressors and sensitivity to antinociceptive effects of acute and chronic morphine. We tested whether early neonatal experience alters sensitivity to effects of morphine on locomotor activity. The subjects were adult rats that had one of the following backgrounds: daily separation from the dam on postnatal days 2-14 for either 3 h (maternal separation (MS)) or 15 min (handled control (H)) or no separation from the dam (non-handled control (NH)). After two consecutive days of baseline activity measurements, subjects were tested daily after SC injections of either morphine (10 mg/kg) or saline for seven days and again on day 10. Beginning five days later, saline and 1.0-10 mg/kg of morphine were tested in all animals. On the baseline days, MS animals had higher horizontal and vertical activity than did NH controls, whereas H animals spent more time in the center of the testing chamber. In MS and H animals but not in NH controls, daily injections of morphine produced progressive increases in all locomotor activity measures, indicative of sensitization (horizontal counts, center time) and tolerance (vertical counts). MS animals with a history of morphine treatment had significantly higher horizontal and vertical activity after a saline injection than did their counterparts with a history of saline treatment, indicative of conditioning. They also exhibited greater locomotor sensitization to 1.0 mg/kg of morphine than did H and NH controls. These results provide further evidence that environmental manipulation in the form of maternal separation early in life results in enduring changes in sensitivity to effects of morphine that could reflect altered endogenous opioid systems.     

The effect of the CS-UCS interval and extinction on place conditioning and analgesic tolerance with morphine

In experiment 1, a CS-UCS interval study of place conditioning and analgesic tolerance with morphine was conducted. Morphine (10 mg/kg i.p.) was administered to separate groups of rats either 2 h prior to, 1 h prior to, immediately prior to, immediately after or 2 h after 30-min confinement in one end compartment of a place conditioning apparatus. A total of three choice tests was given, one after every six morphine injections. A preference for the end compartment contingent upon morphine injection was shown in groups that received morphine prior to end compartment placement. Groups that received morphine after end compartment placement were not different in their preference behavior from groups that received only saline during place conditioning training. A hot-plate test for tolerance to the analgesic effect of morphine was given at the end of all choice testing. All groups that had received morphine during place conditioning training were equally tolerant. These results indicate a dissociation between the analgesic effect of morphine and the effect that produces place preference, since the former was not affected by temporal parameters that did affect the latter. In the second experiment, the effect of extinction on a morphine-induced place preference was studied using extinction procedures that, in contrast to previous studies, equated exposure to both end compartments. Whereas the morphine-induced place preference was undiminished by a 10-day retention period in which animals received saline injections in the home cage, extinction trials during the same period eliminated the place preference. These results provide evidence that morphine-induced place preferences involve associative processes.     

Morphine-induced place conditioning is not confounded by drug-induced alterations in locomotor activity

The influence of locomotor activity and environmental familiarity upon the reinforcing effects of morphine was examined in an unbiased place preference conditioning procedure. Groups of rats were trained to associate one distinctive environment with morphine and another with saline. One group was made tolerant to the locomotor activity effects of morphine by the SC administration of morphine (5.0 mg/kg/12 hr) for four days prior to conditioning. The other group received injections of saline. Administration of morphine, at doses which decreased locomotor activity, resulted in marked preferences for the drug-associated place in saline-treated rats. In contrast, chronic morphine treatment resulted in tolerance to the sedative effects of morphine and an abolition of the morphine-induced place preference. These results indicate that in the place conditioning procedure, measures of reinforcement are not confounded by drug-induced increases in activity.     

Contingent suppression of tolerance to haloperidol: a dose-response analysis

Rats were given injections of haloperidol (HAL; 0.625 or 2.5 mg/kg) either before (Before groups) or after (After groups) access to sweetened milk on alternate days. Controls (Saline groups) were given injections of saline. At biweekly intervals ("test days"), all groups were given pretest injections of the drug in order to monitor the development of tolerance in the After and Saline groups. Rats in the Before groups showed no tolerance to the initial suppression of milk intake. In contrast, rats in the After groups had greater intakes, although the level of intake declined on subsequent test days in the group given the lower dose. Rats in the Saline groups drank less on the test days than any of the other groups, suggesting that sensitization occurred. These results are consistent with previous findings (29) that tolerance to HAL is suppressed following pretest injections of the drug. The degree of suppression appears to be inversely related to the frequency of such injections.     

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.     

Chronic inflammatory pain does not attenuate the development of tolerance to chronic morphine in adult male rats

The overall impact of chronic pain on the response to opioids is ambiguous in the literature, and comparisons between human and animal studies are complicated by vast differences between the manner and dosage of opioids given to humans treated for pain in comparison to rodents as well as a lack of healthy participant studies examining the impact of chronic opioids. The purpose of this study was to evaluate the impact of chronic pain on the development of tolerance to morphine and to assess how the concentration of drug affects this process. Twenty-four hours after the injection of CFA or normal saline in the left hind paw, the level of mechanical hypersensitivity was assessed and animals were randomly assigned to a morphine dose (1, 3 or 8 mg/kg or saline). Morphine was administered by subcutaneous injection twice a day for 5 days. On Day 6, animals were challenged with a single dose of 3 mg/kg morphine prior to formalin testing. Evidence of tolerance was mixed, and the results varied widely among the conditions. Analysis of mean paw withdrawal thresholds indicated that the analgesic efficacy of subcutaneous morphine diminished following repeated dosing. The presence of the chronic inflammatory pain condition during the morphine dosing period produced an increase in formalin pain behaviors compared to saline controls, such that animals given any dose of morphine during the 5-day dosing period showed higher responding to formalin following the 3 mg/kg dose than animals that had received saline injections. These results indicate that chronic pain does influence the development of opioid tolerance, but it does not prevent this phenomenon from occurring as suggested by some researchers.     

Dissociation in the modulatory effects of environmental novelty on the locomotor,analgesic, and eating response to acute and repeated morphine in the rat

Rationale. We have previously shown that environmental novelty can potentiate the activating effects of morphine and the development of sensitization to this effect. Objectives. Our main goal was to determine whether environmental novelty can also modulate the prophagic (time spent eating and food intake; experiment 1) and/or the analgesic (tail-flick test; experiments 2 and 3) effect of morphine, as well as the development of tolerance or sensitization to these effects. Methods. In experiment 1, two groups of rats were administered seven intraperitoneal (i.p.) injections of either saline or morphine (4.0 mg/kg) either in their home cages (home groups) or in a distinct environment (novelty groups). After 7 days of withdrawal, both groups underwent a morphine challenge (4.0 mg/kg, i.p.). In experiment 2, home and novelty rats were administered four doses of morphine (0.0, 2.0, 4.0, and 8.0 mg/kg, i.p.) following a counterbalanced order. In experiment 3, home and novelty rats were administered eight intraperitoneal (i.p.) injections of either saline or morphine (8.0 mg/kg) and then given a morphine challenge (4.0 mg/kg). Results. Environmental novelty enhanced the locomotor activating effect of morphine and the expression of sensitization to this effect (even after a period of withdrawal). Environmental novelty had relatively little effect on morphine-induced eating, and no effect on morphine-induced analgesia. Conclusions. Environmental context can have very different consequences on distinct drug effects as well as on distinct neurobehavioral adaptations to the same drug treatment (e.g., psychomotor sensitization versus analgesic tolerance). Electronic Publication     

Naloxone enhances the expression of morphine-induced conditioned place preference

The present study examined the effects of naloxone on acquisition and expression of morphine-induced conditioned place preference (CPP). Three groups of rats were given morphine (5 mg/kg, SC), both morphine and naloxone (1 mg/kg, SC), or saline paired with a distinctive environment. On alternating days they were given saline paired with another distinctive environment. After four exposures to each environment, the animals were given a preference test in which they had access to both environments simultaneously while under the influence of either naloxone (1 mg/kg, SC) or saline. Morphine-conditioned animals showed CPP evident as an increased amount of time spent in the drug-associated environment relative to saline controls. Rats given both naloxone and morphine during conditioning, and saline on the test day, did not show CPP. In contrast, morphine-conditioned animals given naloxone on the test day showed stronger CPP than morphine-conditioned animals given saline. These findings indicate that naloxone blocks the acquisition, but enhances the expression of morphine-induced CPP. In a separate experiment, the effects of naloxone on locomotor activity were determined during the CPP test. The results indicated that naloxone decreased locomotor activity. In morphine-conditioned animals only, naloxone also produced an increase in the amount of time per entry in the drug-associated environment. The results suggest that naloxone may enhance morphine-induced CPP by decreasing locomotor activity that may otherwise compete with expression of CPP.     

Tolerance to morphine stimulus control: role of morphine maintenance dose

Experiments assessed the development of tolerance to morphine stimulus control during treatment with selected maintenance doses of morphine. Separate groups of rats were trained to discriminate saline and either 3.2 mg/kg or 5.6 mg/kg morphine under fixed-ratio schedules of food delivery. Dose-response functions for generalization of morphine stimulus control were determined before, during, and after repeated treatment with selected doses of morphine. Similar experiments were performed with repeated pentobarbital treatment in order to assess the pharmacological selectivity of tolerance. Repeated treatment with saline, 3.2 mg/kg morphine, or twice daily injections of 17.8 mg/kg pentobarbital produced no tolerance to morphine stimulus control. In contrast, treatment with daily injections of 10 mg/kg or twice daily injections of 10 or 17.8 mg/kg morphine produced a dose-dependent increase in the dose of morphine required for stimulus control. The magnitude of tolerance to morphine stimulus control varied directly with the maintenance dose of morphine and was slightly greater for a lower than a higher morphine training dose. Termination of repeated treatment was followed by a return to initial sensitivity, without additional training. Tolerance to morphine stimulus control was not necessarily accompanied by tolerance to its rate-suppressing effects.     

Repeated administration of nicotine attenuates the development of morphine tolerance and dependence in mice

Clinical use of morphine in pain management is a controversial issue. Both nicotine and morphine are widely abused. So, investigating the interaction between nicotinic and opioid receptors is of great interest to both basic mechanistic and clinical view. We investigated the influence of repeated administration of nicotine on the development of morphine tolerance and dependence. Adult male albino mice were rendered dependent on morphine by subcutaneous (s.c.) injections three times daily for 3 days. Repeated intraperitoneal (i.p.) injection of nicotine (0.001-2 mg/kg) or saline (1 ml/kg) was performed 15 min prior to each morphine injection. Maximal possible effect (MPE%) of morphine (50 mg/kg; s.c.) was used on the fourth day as an index for the development of tolerance. Likewise, to assess the occurrence of dependence in drug-treated mice, naloxone (5 mg/kg; i.p.) was injected 2 h after the last dose of morphine. Repeated nicotine administration significantly attenuated the development of tolerance in a dose-dependent manner whereas it significantly decreased withdrawal jumping behavior in a biphasic profile (V-shape) manner. Furthermore, the central nicotinic receptor antagonist mecamylamine (0.01-0.1 mg/kg; i.p.) neither the peripheral nicotinic receptor antagonist hexamethonium (0.01 and 0.1 mg/kg; i.p.) nor the muscarinic receptor antagonist atropine (2.5-10 mg/kg; i.p.), dose-dependently antagonized both the inhibition of withdrawal jumping as well as increase in MPE% which was produced by repeated nicotine administration (0.1 mg/kg; i.p.). On the other hand, 3 days of solely nicotine treatment resulted in significant jumping behavior precipitated by naloxone after single morphine injection on the test day. The data suggests that the inhibitory effect of nicotine on the morphine tolerance and dependence is mediated by central nicotinic receptors and there is a cross-dependence between nicotine and morphine.     

Long-lasting changes in morphine-induced locomotor sensitization and tolerance in Long-Evans mother rats as a result of periodic postpartum separation from the litter: a novel model of increased vulnerability to drug abuse?

Daily postpartum separations from the litter produce enduring changes in anxiety and sensitivity to the antinociceptive effects of morphine in Long-Evans dams. We tested whether postpartum experience alters sensitivity to the effects of morphine on locomotor activity. Dams were tested 4-6 weeks after their pups were weaned, and had one of the following backgrounds: daily separation from the litter on postpartum days 2-14 for either 3 h (prolonged separation-LS) or 15 min (brief separation-BS), or no separation (nonhandled control-NH). After 2 consecutive days (B1-2) of baseline activity measurements, subjects were tested daily after s.c. injections of either morphine (10 mg/kg) or saline for 7 days and again on day 10. Beginning 5 days later, saline and 1.0-10 mg/kg of morphine were tested in all dams. On B1, LS and BS dams habituated slower than NH controls, yielding higher horizontal counts. LS dams failed to habituate across baseline days and were more active than other dams on B2. Sensitization, a progressive increase in horizontal activity, was more rapid and robust in LS and BS dams compared to NH animals. LS was the only group that developed tolerance to morphine-induced decreases in vertical activity. In LS dams with the history of morphine treatment, injection of saline resulted in higher horizontal activity and center time compared to saline-treated counterparts, indicative of conditioning. Among animals with a history of saline treatment, LS dams were more sensitive to morphine challenges than BS and NH dams. As a result of the robust and long-lasting increases in the ability of morphine to induce behavioral sensitization in litter-separated dams, periodic postpartum separation may represent a new animal model of increased vulnerability to substance abuse.     

The acoustic startle response as a measure of behavioral dependence in rats

A series of experiments was conducted to assess the sensitivity of the acoustic startle response to chronic morphine administration and naloxone-precipitated withdrawal. Rats were implanted with two subcutaneous pellets containing either 75 mg each of morphine or containing only placebo. In experiment 1, withdrawal induced by 0.05–0.2 mg/kg naloxone dose-dependently decreased the magnitude of the startle response. Physical dependence was confirmed by a naloxone-induced acute weight loss seen in morphine-implanted rats, but naloxone had no effect on startle or body weight in nondependent animals. In experiment 2, a modified procedure with fewer trials per session and fewer test days was employed. Naloxone (0.2 mg/kg) given 4–5 days after implantation induced large startle-response decreases in morphine-dependent rats while having no effect in placebo-implanted rats. Post-naloxone saline tests revealed no significant differences in startle between morphine and placebo groups. Startle scores were significantly higher in morphine-implanted rats than in placebo rats during a saline test given 3 days following pellet implantation. In a separate group of animals, however, acute IP injections of morphine from 0.3–10 mg/kg had no significant effect on startle amplitude. The effect of repeated pairings of withdrawal with the startle environment was assessed in experiment 3. Morphine-dependent rats startled significantly less if naloxone injections were given before the startle session than if they were administered 4 h later. Conditioned withdrawal effects, expressed during a final test session when all rats received saline, were observed for the body-weight measure but not for the startle response. These results suggest that the acoustic startle response may be a useful objective measure in evaluating physical dependence produced by substances of abuse.     

Effects of test conditions on the outcome of place conditioning with morphine and naltrexone in mice

 Drug administration during test trials can increase the expression of place conditioning, offering an opportunity to determine the specificity of this enhanced response. Prior to training, Swiss-Webster mice spent similar durations in each of the distinctive compartments of a two-compartment box during three 900-s tests. During a 4-day conditioning period, daily injections of morphine (5–20 mg/kg, SC) or vehicle were differentially paired with one of two compartments of the box using an unbiased place conditioning procedure. Post-conditioning tests were conducted 2 and 3 days after the last conditioning day. Mice pre-treated during post-conditioning tests with vehicle did not show significant preference for the morphine-paired compartment when conditioned with morphine. Pretreatment with morphine (2.5–30 mg/kg, SC) led to a dose-dependent increase in time spent in the morphine-paired compartment. Post-conditioning tests in other groups of mice were conducted with heroin (0.1–3 mg/kg), fentanyl (0.01–0.3 mg/kg), cocaine (10–30 mg/kg) and pentobarbital (10–30 mg/kg), and results suggested that none of the tested drugs facilitated the expression of the morphine-conditioned place preference. In another experiment, naltrexone (0.1–10 mg/kg, SC) was administered as the conditioning drug. When tested with naltrexone (0.1–10 mg/kg), there was a dose-dependent avoidance of the naltrexone-paired compartment. Overall, the present data indicated that: (1) failure to exhibit place preference or place aversion when tested in a drug-free state does not imply the failure of conditioning procedure; and (2) effects of the morphine cue reinstatement during the post-conditioning tests appeared to be related to the unique pharmacological profile of the morphine stimulus. Received: 28 November 1997 / Final version: 23 July 1998