Acute delta- and kappa-opioid agonist pretreatment potentiates opioid antagonist-induced suppression of water consumption.

The primary objective of this study was to determine whether pretreatment with kappa- and delta-opioid agonists potentiates naltrexone-induced suppression of water consumption following 24h of deprivation. This study also examined the temporal effects of agonist-induced antinociception using the tail-flick and hot-plate tests. Adult male Sprague-Dawley rats were water deprived 20 h and then given an injection (s.c. or i.c.) of an opioid agonist or saline. Drugs included the mu-opioid agonists morphine and DAMGO ([d-Ala2,NMePhe4,Gly-ol5]-enkephalin), the kappa-opioid agonists spiradoline, bremazocine, and U69,593, and the delta-opioid agonists BW 373U86 and DPDPE ([D-Pen2, D-Pen5]-enkephalin). Three hours and forty-five minutes later, animals received a single dose of naltrexone (0.1-30 mg/kg, s.c.) or saline. Fifteen minutes later, animals were allowed free access to water for 30 min. For the tail-flick and hot-plate tests, animals were given a single injection of agonist and tested in both procedures every 30 min for up to 2h, then hourly up to 6h post-injection. Naltrexone dose-dependently suppressed fluid consumption 24h after deprivation. The effects of naltrexone on drinking were potentiated following pretreatment with at least one dose of the agonists tested except BW 373U86. With the exception of BW 373U86, DAMGO, and DPDPE, all of the opioid agonists produced significant antinociception in the hot-plate test. Only BW 373U86 failed to have an antinociceptive effect in the tail-flick test. By 4h after treatment, drug-induced antinociception had largely waned, suggesting the potentiation of naltrexone-induced drinking suppression was not a result of a direct interaction with the agonists. In conclusion, kappa-opioid and delta-opioid receptors appear to contribute to the manifestation of acute opioid dependence, albeit to a lesser degree than mu-opioid receptors.

Early postnatal stress alters place conditioning to both mu- and kappa-opioid agonists.

Clinical literature has established a link between early childhood incidents of neglect and trauma and adult problems with substance abuse. In rats, such early life stress has been modeled using a maternal separation (MS) paradigm in which rat pups were removed from their mothers for a few hours daily during the first two postnatal weeks. In this study, we used the MS model to investigate the effects of early postnatal stress on place conditioning to both mu- and kappa-opioid agonists in male and female Long-Evans rats. Offspring of both rearing conditions [MS or nonhandled (NH)] were conditioned using a biased procedure to saline, the mu-opioid agonist morphine (3.0, 5.6, and 10 mg/kg s.c.), or the kappa-opioid agonist spiradoline (0.3, 1.0, and 3.0 mg/kg) for 3 days, followed by a drug-free place-conditioning test 24 h later. Saline was administered in the morning, 30 min before confinement in one compartment, whereas morphine or spiradoline was administered in a similar manner 6 h later in the opposite compartment. MS offspring spent significantly more time in the morphine-paired compartment than NH offspring, indicating a greater place preference for the mu-opioid agonist. In the case of spiradoline, NH offspring spent significantly less time in the spiradoline-paired compartment, indicating a greater aversion to the kappa-opioid agonist in these animals than in MS offspring. These findings indicate that early postnatal stress can significantly alter the rewarding or aversive value of mu- and kappa-opioid agonists when measured using place conditioning.

Periadolescent male but not female rats have higher motor activity in response to morphine than do adult rats.

Little has been done to investigate the effects of opioid exposure during adolescence. First we determined behavioral differences in response to acutely administered morphine between periadolescent and adult male and female rats. Second, we determined the impact of age of morphine exposure on sensitivity to morphine-induced locomotion later in life. For the acute morphine studies, antinociceptive responses were assessed using cumulative morphine dosing (0.5-12 mg/kg) followed by a time course after the last morphine injection (

The CRF1 receptor antagonist, R121919, attenuates the severity of precipitated morphine withdrawal.

Corticotropin-releasing factor (CRF) regulates the hypothalamic-pituitary-adrenal axis, coordinates the mammalian stress response, and acting primarily via the CRF(1) receptor, has been strongly implicated in the pathophysiology of depression and anxiety. Furthermore, the behavioral and autonomic activation that occurs following withdrawal in drug dependent animals resembles the mammalian stress response. Concordant with this view is evidence of enhanced CRF transcription, release and activity following withdrawal from several drugs of abuse. Conversely, CRF receptor antagonists have been demonstrated to reduce the severity of many drug withdrawal symptoms, implicating a specific role for activation of CRF neurons in mediating the anxiogenic and stress-like reactions observed during withdrawal. To extend these findings, we investigated whether pretreatment with a selective CRF(1) receptor antagonist, R121919, is capable of similarly decreasing the autonomic, behavioral and neuroendocrine activation observed following precipitation of morphine withdrawal in dependent rats. The results indicate that pretreatment with R121919 attenuates the global severity of the precipitated morphine withdrawal syndrome as measured by the Gellert-Holtzman scale. In addition, rats pretreated with R121919 prior to precipitation of morphine withdrawal demonstrated decreased hypothalamic-pituitary-adrenal axis activation, as measured by plasma ACTH concentrations, and decreased early expression of the CRF gene in the paraventricular nucleus of the hypothalamus, as measured by CRF heteronuclear RNA. These findings suggest that activation of CRF neuronal systems via the CRF(1) receptor may be one element of the neurobiological mechanisms activated during drug withdrawal and that CRF(1) receptor antagonists may have a potential therapeutic role in the treatment of human drug withdrawal syndromes.

Maternal separation alters ICSS responding in adult male and female rats, but morphine and naltrexone have little affect on that behavior.

Prior research has provided evidence that the early postnatal environment can have long lasting effects on both the physiology and behavior of offspring. This is modeled in rats by using a maternal separation paradigm in which pups are separated from their mother for a few hours daily during their first two postnatal weeks. While this model has been used extensively to study stress effects and anxiety, less research has been done to examine how these separations affect measures of reward and reinforcement in adulthood. The current study investigated the impact of maternal separation (MS) on intracranial self-stimulation (ICSS) maintained responding in male and female offspring, and the effects of morphine (0.3-3.0 mg/kg) and naltrexone (0.1-10 mg/kg) on that responding. Rearing condition (MS or non-handled, NH) significantly altered response rates during acquisition in both sexes, with NH offspring exhibiting the highest rates. Group differences in baseline responding on a progressive ratio (PR-2) schedule of reinforcement were evident only in females, with MS females having response rates 50% lower than NH females. Neither morphine nor naltrexone differentially affected either rearing group. Sex impacted NH offspring: males acquired responding more readily, but females had higher response rates and breakpoints during all other phases of the experiment. In MS offspring, no sex differences were observed during acquisition, but during all other phases males had higher response rates and breakpoints than females. These results indicate that maternal separation during the first two postnatal weeks can have long-term effects on responding for ICSS, but these effects do not appear tied to endogenous opioid systems in the lateral hypothalamus.

Enhanced sensitivity to naltrexone-induced drinking suppression of fluid intake and sucrose consumption in maternally separated rats.

Early-life stress has been identified as a risk factor in the development of a host of disorders, including substance abuse; however the link between early postnatal stress and changes in measures of reward has not been thoroughly researched. The current study had two main objectives: 1) to determine the impact of maternal separation (an animal model of early-life stress) on the consumption of 10% and 2.5% sucrose solutions by Long-Evans rat dams and male and female offspring, and 2) to determine the effect of the opioid antagonist naltrexone (0.1-3.0 mg/kg) on drinking by each of those groups. Dam-pup separations occurred for varying lengths of time during the first two postnatal weeks. In Experiment 1, a two-bottle choice test (sucrose solution vs. water) was administered across five days to both nonhandled (NH) and maternally-separated (MS) offspring as adults and to dams 2-4 weeks post-weaning. In Experiment 2, naltrexone was administered prior to two-bottle choice tests. MS males and the dams of MS litters exhibited increased intake of total fluid and sucrose solutions, whereas results from females were less consistent. Naltrexone elicited a greater decrease in fluid intake and sucrose intake in male MS offspring compared to male NH offspring. These results indicate that early postnatal stress alters both sucrose consumption, a non-drug measure of reward, and apparently the brain opioid systems that mediate naltrexone-induced drinking suppression.

Locomotor response to novelty as a predictor of reactivity to aversive stimuli in the rat.

In an animal model for vulnerability to drug abuse, animals that exhibit greater motor activity in a novel environment (high responders; HR) are more sensitive to drugs of abuse and are more likely to self-administer these drugs compared to less reactive animals (low responders; LR). In the light of clinical evidence on comorbidity between drug abuse and mood disorders, we used this model to investigate whether individual differences in locomotor reactivity to novelty are related to anxiety- and depression-like responsiveness using male Sprague-Dawley rats. Animals were categorized as HR and LR based on motor responses to novelty during a 30-min session. Anxiety-like reactivity was then measured using the elevated plus-maze, the defensive withdrawal test and acoustic startle-induced ultrasonic vocalization test. Depression-like reactivity was measured by the forced swim test. HR rats showed less anxiety-like behavior in the elevated plus-maze and defensive withdrawal tests than LR, but the opposite was true in the acoustic startle-induced vocalization test. In response to a series of loud acoustic stimuli, HR rats were faster to begin vocalizing and did so for a longer duration compared to LR. There were only minor differences between LR and HR rats in the forced swim test. These data suggest that an HR/LR model can be used to study a link between vulnerability to drug abuse and anxiety-like reactivity. The exact nature of this link depends upon the model of anxiety used and may reflect the heterogeneous nature of anxiety-like reactivity in the rat.

Neonatal stress and litter composition alter sucrose intake in both rat dam and offspring.

The early postnatal environment can have long lasting effects on the physiology and behavior of both mother and offspring. A great deal of evidence indicates that stress during this time period is a risk factor for the future development of a multitude of disorders including substance abuse. The maternal separation paradigm is used to model such stress in rats. The current study evaluated the effects of maternal separation and litter composition on sucrose consumption, a non-drug measure of reward, in both male Long-Evans rat offspring and mother. On postnatal day 2, rats were cross-fostered, placed in single-sex or mixed-sex litters, and subsequently stressed by daily dam-pup separations during the first two postnatal weeks. The length of the separation (15 min, 1 h, or 3 h) was randomly assigned to each day. A two-bottle choice test was given to maternally separated and nonhandled offspring when they were adults, and to dams 2-4 weeks after weaning. Intake of a 10% sucrose solution or water was compared for 1 h daily across five consecutive days. Rearing condition had a profound effect on total fluid intake and sucrose solution intake by both offspring and dam, with the separated offspring and dams generally consuming a greater amount of total fluid and 10% sucrose solution than their nonhandled counterparts. Litter composition also affected consumption, with the offspring from maternally separated mixed-sex litters consuming more total fluid and 10% sucrose solution than offspring from maternally separated single-sex litters. These results indicate that early postnatal and postpartum stress can lead to changes in sucrose consumption, a non-drug measure of reward, indicating that such stress may alter the underlying brain reward mechanisms.

Modeling the onset of drug dependence: a consideration of the requirement for protein synthesis.

It has been proposed, with some supporting evidence, that development of opiate tolerance and dependence requires protein synthesis. However, a quantitative, biologically based model within which to analyse and support the data has been lacking. Utilizing such a framework or model, we recently compared the time course of onset of opiate dependence in laboratory animals, with the mathematical time course of general changes in protein levels. Not only did the time course of onset of dependence parallel the time course of increasing levels of a protein, but also the half-life of the putative protein required by the model was very similar to those of many brain proteins. In this study, we have more extensively tested the model by producing and examining a much more detailed and surprisingly complex time course of the onset of dependence. Applying the protein synthesis time course model to the data suggested the presence of two distinct components of dependence, an early transient component and a later long-lasting component. These components appear to correspond to acute and chronic dependence, respectively. The protein synthesis hypothesis more readily applies to the chronic dependence portion. Because consideration of the model can generate components that correspond to accepted and well-known components of dependence, both the utility of the model as well as the hypothesis that opiate dependence at least partially requires protein synthesis are supported. It is also possible that individual components of the withdrawal syndrome have individual and unique rate limiting mechanisms. In any case, time course analysis may be helpful in revealing underlying mechanisms of change.

Periadolescent morphine exposure alters subsequent behavioral sensitivity to morphine in adult rats.

Little research has been conducted investigating the long-term impact of opioid exposure during adolescence. These experiments were conducted to determine the behavioral effects of morphine exposure during periadolescence (postnatal days 30-32) versus young adulthood (postnatal days 65-67) on subsequent sensitivity to morphine. Male Sprague-Dawley rats were treated with three days of saline (S-S-S), one day of 10 mg/kg morphine followed by two days of saline (M-S-S), or three days of morphine (M-M-M). Unlike adult-treated counterparts, periadolescent M-M-M-treated rats showed greater locomotor response to morphine compared to S-S-S or M-S-S cohorts five weeks after treatment, suggesting age- and exposure-dependent differences in opioid sensitivity.

Naltrexone-induced conditioned place aversion following a single dose of morphine in the rat.

Limited preclinical research has been conducted investigating the motivational or "affective" properties of withdrawal from acute opioid dependence following a single morphine exposure. Therefore, the purpose of the present study was to pharmacologically characterize the motivational properties associated with naltrexone-precipitated withdrawal after a single injection of morphine using place conditioning. Conditioned place aversion was assessed using a biased two-compartment apparatus and procedure. Adult male Sprague-Dawley rats were given 15 min free access to the entire apparatus on day one to determine initial preferences. Beginning on the second day, combinations of either saline or morphine (1.0-10 mg/kg, s.c.) followed by naltrexone (0.003-3.0 mg/kg, s.c.) 3.75 h later were administered. Rats were then immediately confined to one compartment for 30 min. The next day, rats received the alternative treatment and were confined to the opposite compartment. Twenty-four hours later animals were tested again for 15 min while they had access to the entire apparatus. Morphine followed by naltrexone conditioned significant place aversion (CPA) with just one pairing. This effect was a function of the naltrexone and morphine doses. CPA was also dependent on morphine pretreatment time, with significant aversion only occurring 4 h after morphine pretreatment. Finally continuous morphine administration followed by a single injection of naltrexone resulted in CPA. These data extend the range of behavioral effects associated with antagonist-precipitated withdrawal from acutely administered morphine and suggest that place conditioning is an effective model in assessing motivational aspects of withdrawal from acute opioid dependence in rats.

Caffeine induces differential cross tolerance to the amphetamine-like discriminative stimulus effects of dopaminergic agonists.

The purpose of this study was to determine if caffeine induces cross tolerance to the amphetamine-like discriminative stimulus effects of dopaminergic drugs that act through distinct mechanisms (e.g., release, uptake inhibition, direct activation of dopamine D(1)- or D(2)-family receptors). Rats were trained to discriminate 1.0 mg/kg d-amphetamine from saline in a two-choice discrete-trial procedure. Stimulus-generalization curves were generated by cumulative dosing for d-amphetamine (0.1-1.0 mg/kg), methylphenidate (0.3-5.6 mg/kg), SKF 81297 (0.3-3.0 mg/kg), and R-(-)-propylnorapomorphine (NPA; 0.001-1.78 mg/kg), as well as for caffeine (3.0-56 mg/kg); curves were re-determined after twice daily injections of caffeine (30 mg/kg) for 3.5 days. The rats generalized dose dependently to the four dopaminergic drugs, but only to a limited extent to caffeine. Twice daily injections of caffeine induced significant cross tolerance (i.e., increased ED(50)) to the amphetamine-like discriminative effects of methylphenidate and SKF 81297, attenuated non-significantly the effects of NPA, and did not alter the effects of amphetamine. Thus, caffeine produces differential cross tolerance to the amphetamine-like discriminative effects of dopaminergic drugs, a phenomenon in which the dopamine D(1) receptor appears to have an important role.

Discriminative stimulus effects of acute morphine followed by naltrexone in the squirrel monkey: a further characterization.

The discriminative stimulus effects of acute morphine followed by naltrexone have been described previously in nonhuman primates. The purposes of this study were to 1) extend the pharmacological characterization of the discrimination by testing mu-opioid agonists other than morphine and opioid-like compounds other than naltrexone and 2) to examine further the relationship between agonist pretreatment time and manifestation of the cue produced by morphine followed by naltrexone. Subjects were trained to discriminate 1.7 mg/kg morphine --> 0.1 mg/kg naltrexone (MOR --> NTX) versus saline followed by 0.1 mg/kg naltrexone. When combined with 0.1 mg/kg naltrexone, all agonists tested, save buprenorphine, meperidine, and nalbuphine, produced dose-dependent increases in MOR --> NTX-appropriate responding, culminating in criterion levels of responding. Comparing agonist ED50 values revealed a rank order of potency of etorphine >> fentanyl >> levorphanol > heroin > or = methadone > or = nalbuphine > or = morphine. ED50 values for buprenorphine and meperidine could not be calculated. MOR --> NTX-appropriate responding after doses of agonist that produced criterion or near criterion levels of responding was also a function of naltrexone dose. After morphine pretreatment, diprenorphine and nalorphine, but not buprenorphine, dose-dependently substituted for naltrexone. The MOR --> NTX discrimination also depended upon the interval between morphine and NTX administration. Finally, 1-h pretreatment with morphine and etorphine, but not buprenorphine, followed by naltrexone generalized to 4 h MOR --> NTX. These results suggest a minimum efficacy requirement of acutely administered agonists together with the naltrexone training dose for stimulus control of behavior. However, in some cases this requirement can be overcome with higher doses of naltrexone.

Individual differences in locomotor reactivity to a novel environment and sensitivity to opioid drugs in the rat. I. Expression of morphine-induced locomotor sensitization.

RATIONALE: Vulnerability for development of substance abuse is often associated with a "sensation-seeking" or "thrill-seeking" phenotype. In an animal model, rats more reactive in a novel environment (high responders, HR) are more sensitive to stimulant/reinforcing effects of amphetamine and are more likely to self-administer this drug, than are less reactive animals (low responders, LR). OBJECTIVE: We tested whether HR and LR also differ in sensitivity to effects of morphine on locomotor activity. METHODS: Male Sprague-Dawley rats were categorized as HR or LR based on motor responses to novelty (sorting day; S). After 1 day (B) of baseline activity measurements, subjects were tested daily after SC injections of either morphine (10 mg/kg) or saline for 7 days and again on day 10. Beginning 5 days later, four daily injections of saline and 1.0-10 mg/kg morphine were tested in all animals. RESULTS: LR and HR were similar in the onset and overall magnitude of sensitization and tolerance following daily morphine administration. HR were more sensitive than LR to locomotor stimulant effects of acute morphine. However, LR had more robust and persistent context-specific increases in activity due to conditioning than did HR, and expression of sensitization was apparent in all behavioral variables. CONCLUSIONS: These results provide further evidence that phenotypic differences between HR and LR may, in part, be associated with differences in the endogenous opioid systems. Differences in sensitivity to acute versus repeated morphine suggest that at least in relation to opioid drugs, these phenotypes may reflect different aspects of drug vulnerability rather than simply the presence or absence of it.

Individual differences in locomotor reactivity to a novel environment and sensitivity to opioid drugs in the rat. II. Agonist-induced antinociception and antagonist-induced suppression of fluid consumption.

RATIONALE: In an animal model for vulnerability to drug abuse, rats that are more reactive to a novel environment (high responders, HRs) are more sensitive to behavioral effects of psychostimulants than are less reactive rats (low responders, LRs). In a companion article, we reported that HRs and LRs differ in sensitivity to morphine-induced locomotor sensitization. OBJECTIVE: We tested whether LRs and HRs also differ in sensitivity to opioid-induced antinociception and opioid antagonist-induced suppression of fluid consumption. METHODS: LRs and HRs were categorized based on motor responses to novelty during a 30-min session. Responses to nociceptive stimuli of varied intensities were measured using the tail-flick and hot-plate tests alone or following cumulative doses of morphine (1.0-12 mg/kg), buprenorphine (0.025-0.6 mg/kg), or etorphine (0.25-6.0 microg/kg). Potential changes in endogenous opioid-mediated reward systems were tested using naltrexone-induced (0.01-30 mg/kg) suppression of drinking either water following 24-h deprivation or sweetened condensed milk in a non-deprived state. These effects were further examined following 2 weeks of daily access to sweetened condensed milk. RESULTS: At the lowest stimulus intensity tested, HRs had significantly shorter tail-flick response latencies than LRs. Additionally, HRs were less responsive to cumulative doses of morphine than LRs. There were no overall group differences in the hot-plate test. Following 2 weeks of daily access to sweetened condensed milk, HRs were more sensitive to naltrexone-induced suppression of consumption. CONCLUSIONS: Under the proper conditions, differences in sensitivity to opioid drugs between HRs and LRs at least partially extend to antinociceptive and appetitive reward systems and are suggestive of more extensive differences in phenotype. As with the effects of repeated morphine exposure on locomotor activity, the effect of repeated exposure to appetitive reward associated with sweetened milk appears to be more robust in LRs than HRs.

In rats, acute morphine dependence results in antagonist-induced response suppression of intracranial self-stimulation.

RATIONALE: Lower (0.001-1.0 mg/kg) doses of the opioid antagonist naltrexone produce few behavioral effects in otherwise drug-free rats responding for ICSS, but reduce response rates by up to 75% after a single dose of morphine. OBJECTIVES: The present study represents an effort to verify that other opioid antagonists produce this acute opioid dependence effect, and to characterize their relative pharmacological profiles. METHODS: We implanted bipolar electrodes in the lateral hypothalamus of adult male rats, and then trained them to lever-press on an "autotitration" ICSS schedule, where responding on a "reset" lever allows the rat to control the frequency of stimulation; performance stabilized at approximately 1.5 responses/s. RESULTS: During twice-weekly test sessions, cumulative doses of five of seven opioid antagonists produced significant response rate decreases (30-80%) in saline-pretreated rats; nalorphine (ED25=15.6 mg/kg)> naltrexone (ED25=13.1 mg/kg)>naloxone (ED25=7.3 mg/kg)>levallorphan (ED25=13.96 mg/kg)>(-)cyclazocine (ED25=0.028 mg/kg). A single MOR pretreatment (10 mg/kg, 4 h) significantly enhanced the rate-decreasing effects of six of the seven agonists tested; by 10-fold (-) cyclazocine>13-fold (nalorphine)>93-fold (levallorphan)>972-fold (naloxone)>2190-fold (naltrexone). The pure non-selective antagonist diprenorphine potently decreased rates after MOR pretreatment (ED25= 0.01 mg/kg), but did not after saline pretreatment. The mixed opioid agonist-antagonist drug nalbuphine (1.0-30 mg/kg) did not affect responding after either saline or MOR. CONCLUSIONS: Antagonists with a high affinity for, and a lack of intrinsic activity at, the micro-opioid receptor precipitate the greatest behavioral changes in rats acutely dependent on MOR.

Discriminative stimulus effects of acute morphine followed by naltrexone in the squirrel monkey.

RATIONALE: The discriminative stimulus effects of a combination of acute morphine followed by naltrexone have been described in rats. OBJECTIVE. The purpose of this study was to extend observations to a non-human primate. METHODS: Eight squirrel monkeys were trained in a discrete-trial avoidance/escape procedure to discriminate morphine (1.7 mg/kg, IM, 4 h) followed by naltrexone (0.1 mg/kg, IM, 0.25 h) (MOR-->NTX) versus saline (1.0 ml/kg, IM, 4 h) followed by naltrexone (0.1 mg/kg, IM, 0.25 h) (SAL-->NTX). RESULTS: Seven subjects acquired the discrimination in an average of 108+/-14 sessions. MOR-->NTX-appropriate responding increased as an orderly function of increasing dose of morphine (0.56-1.7 mg/kg) and of naltrexone (0.01-10 mg/kg). The discrimination was also dependent upon interval between morphine and naltrexone administration. The MOR-->NTX cue was fully generalized to the combination of levorphanol (0.3 mg/kg) followed by naltrexone, but not to the non-opioid stereoisomer of levorphanol, dextrorphan (0.3 and 3.0 mg/kg) or the kappa-opioid-receptor-selective agonist U69,593 (0.3 mg/kg) followed by naltrexone. Naltrexone administered 15 min before morphine dose-dependently blocked MOR-->NTX-appropriate responding. CONCLUSIONS: This is the first non-rodent study of the discriminative effects of MOR-->NTX. MOR-->NTX produces a unique interoceptive stimulus that is pharmacologically selective, requires occupation of opioid receptors, presumably mu, for some minimum period of time, and is reversible. This discrimination procedure might provide new insights into the early drug-receptor interactions that underlie the development of physical dependence upon morphine-like drugs.

Discrimination of a single dose of morphine followed by naltrexone: substitution of other agonists for morphine and other antagonists for naltrexone in a rat model of acute dependence.

Rats were trained to discriminate 4-h pretreatment with 10 mg/kg morphine and 15-min pretreatment with 0.3 mg/kg naltrexone (morphine-->naltrexone) from pretreatment with saline and 0.3 mg/kg naltrexone (saline-->naltrexone). The discrimination seems to derive from interoceptive stimuli from antagonist-precipitated withdrawal from acute morphine dependence. The purpose of this study was to extend pharmacological characterization of the discrimination by testing opioid agonists other than morphine and antagonists other than naltrexone. Of seven mu-opioid agonists tested in place of morphine, only two (heroin and levorphanol) substituted completely for it; trials completed on the morphine-->naltrexone-appropriate lever increased as a function of agonist and naltrexone dose. Agonists with intrinsic efficacy higher (etorphine, fentanyl, and methadone) or lower (buprenorphine and meperidine) than that of morphine substituted only partially. However, when naltrexone was administered during continuous infusion of fentanyl or methadone via s.c. osmotic pump, rats responded as if they had received morphine-->naltrexone; discriminative responding correlated with global withdrawal scores. Rats responded primarily on the saline-->naltrexone-appropriate lever when naltrexone was administered after pretreatment with dextrorphan, the dextrorotatory isomer of levorphanol, or kappa-opioid agonists (5-alpha,7-alpha,8-beta)-(+)-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro(4,5)dec-8-yl]-benzeneacetamide (U69,593) and spiradoline. Antagonists with no intrinsic efficacy at mu-opioid receptors (naloxone and diprenorphine) substituted completely for naltrexone, whereas those with some efficacy (nalorphine and levallorphan) substituted partially. Thus, morphine-->naltrexone-like stimulus control of behavior by drugs administered acutely requires pretreatment with certain mu-opioid agonists and a pure antagonist, is independent of agonist efficacy, and is stereoselective. Interoceptive stimuli from naltrexone-precipitated opioid withdrawal are more similar across morphine-like agonists during chronic dependence than they are during acute dependence.

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.

Changes in urination/defecation, auditory startle response, and startle-induced ultrasonic vocalizations in rats undergoing morphine withdrawal: similarities and differences between acute and chronic dependence.

In drug-free subjects, a single dose of morphine followed by an opioid antagonist a few hours later results in signs of a withdrawal syndrome, suggesting a state of physical dependence. Increased urination/defecation, altered startle, and ultrasonic vocalizations (USV) are some signs of the withdrawal syndrome in rats chronically dependent on morphine. We investigated whether naltrexone stimulates urination/defecation and alters startle and USV in male rats that were pretreated with only a single dose of morphine and compared these indices to the ones of chronic dependence. Separate groups of rats were pretreated with either a single dose (10 mg/kg) or with a continuous s.c. infusion of morphine via an osmotic pump. Naltrexone (0.01-1.0 mg/kg) was administered 2 to 6 h after the single dose of morphine and on days 7 to 11 of the infusion. Immediately after the naltrexone injection subjects were placed in sound-attenuating boxes to record startle and USV and to collect urine/feces. Subjects chronically exposed to morphine also were tested during spontaneous withdrawal 3 to 24 h after pump removal. Naltrexone increased urination/defecation in subjects pretreated with morphine either chronically or acutely; it increased startle and USV in acutely dependent rats but decreased them in chronically dependent rats. In the latter group, changes in the four variables during spontaneous withdrawal were qualitatively similar to those during precipitated withdrawal but smaller in magnitude. Differences in withdrawal signs between acute and chronic dependence suggest that the neural substrates that mediate those particular components of the withdrawal syndrome are affected differently in the two states of dependence.