Naloxone prevention of morphine LDR curve flattening associated with high-dose tolerance

Male wistar rats, previously made tolerant to morphine by at least 3 weeks of daily intraperitoneal (IP) injections of 20 mg/kg morphine-SO₄ (MS), were then given 200 mg/kg MS daily for 4 or 5 days. Tail immersion tests of antinociception, carried out before and after the 200 mg/kg MS treatment, indicated that the additional morphine treatment was followed by a large further decrease in opiate sensitivity, characterized by decreased slope of the log-dose/response curve (LDR curve flattening). The further decrease in opiate sensitivity was substantially reduced by naloxone-HCl (IP) in a dose of 10 mg/kg given 30 min before and 8 h after the 200 mg/kg MS injections, or a dose of 4 mg/kg given 45 min after the MS. It was concluded that LDR curve flattening produced by high doses of MS is mediated by specific opiate receptors, and is a true expression of a high degree of opiate tolerance in the intact rat.     

Log dose/response curve flattening in rats after daily injection of opiates

Rats injected (IP) daily with 0, 20, and 200 mg/kg morphine-SO₄ for 25–49 days experienced log dose/response (LDR) curve flattening (decrease in slope and/or maximum response) for analgesia (tail immersion test) produced by etorphine-HCl injected IP or intracerebroventricularly (ICV), and for latency to maximum rectal temperature increase produced by IP etorphine. Rats treated similarly with 0, 50, and 500 g/kg etorphine-HCl for 32 days exhibited LDR-curve flattening for analgesia produced by etorphine and morphine (IP). In addition, a profound body weight loss produced by high-dose morphine treatment (200 mg/kg) was found not to be involved in flattening, since similar body weight decreases produced by food restriction in 0 and 20 mg/kg rats did not have this effect. Flattening, however, may be due to a rapidly acquired and rapidly lost within-session (acute) tolerance. When flattening was not seen at short intervals after IP or ICV test etorphine doses, flattening was seen when rats were retested at longer test intervals. Forty-eight hours after cessation of chronic etorphine treatment, flattening of the etorphine analgesia LDR curve was lost, but parallel shift was unaffected. Similarly, 200 mg/kg morphine-treated rats lost morphine tolerance more rapidly than 20 mg/kg-treated rats during the first 12 days after the last treatment injection. Subsequently, however, levels of the analgesia and the amounts of tolerance loss were comparable in both chronically treated groups. The data support the notion that chronic tolerance reflects an enhancement or prolongation of acute tolerance.     

Quantitative relationships among measures of morphine tolerance and physical dependence in the rat

The influence of treatment dose on a number of characteristics of opiate tolerance was studied in male Sprague-Dawley rats treated with daily intraperitoneal (IP) injections of morphine sulfate. Zero, 7.5, 15, 25 or 45 mg/kg/day was given for 34 consecutive days and the degree of morphine effect on four different tests was periodically assessed. Dose-related effects on tailflick latency (tail immersion test, 1.5 hr post injection), swimming test (2 hr post injection), and body weight gain revealed the development of tolerance; there was a non-dose-related hyperthermia (1.5 hr post injection) to which rapid sensitization occurred. All changes reached asymptote and the rate and extent of change varied with the test. Plots of log response vs test day for tailflick and swimming indicated an early steep component and a later less steep component of decline. Subsequent testing indicated that the log-dose/response (LDR) curves for tailflick latency and time to maximum hyperthermia shifted to the right by an amount dependent on the treatment dose; there was no change in the curve for hyperthermia duration. In high dose groups no further shift occurred, but the tailflick LDR curves became flattened. The tailflick LDR curve changes were replicated in rats treated for 24 days with 0, 8, 24, 48, 96, or 240 mg/kg/day. Subsequently, a constellation of withdrawal signs precipitated by naloxone HCl (1 mg/kg, IP) was measured. On the basis of the relation between treatment dose and the magnitude of the various measures, there was a parallel between analgesia tolerance and some, but not all, signs of physical dependence.     

Effects of memantine on estrogen-dependent acute tolerance to the morphine analgesia in female rats

Both human and animal studies suggest that there are sex differences in responding to noxious stimulation as well as in effects of opiate analgesic drugs. Development and/or expression of tolerance to opiate analgesia are also affected by the hormonal status of the experimental subjects. The present study aimed to compare acute tolerance to morphine in cycling and ovariectomized female rats and to evaluate the effects of N-methyl-D-aspartate (NMDA) receptor channel blocker memantine as well as 17-beta-estradiol on tolerance development using the tail-flick test. Acute tolerance to morphine analgesia was observed as a substantial reduction in the response to a test dose of morphine (10 mg/kg) given 6 h after the tolerance-inducing dose (10 mg/kg). Significant acute tolerance was observed in proestrous female rats and was prevented by memantine (3 or 10 mg/kg) treatment. Ovariectomized rats did not demonstrate tolerance to morphine analgesic effects but chronic estradiol administration (5 microg/day, 5 days) reinstated induction of tolerance. Both estrogen receptor modulator tamoxifen (5 mg/kg/day, 5 days) and memantine (3 mg/kg/day, 5 days) prevented estradiol-induced tolerance in ovariectomized rats. Thus, estrogens were found to play a key role in induction of acute tolerance to morphine antinociception. Estradiol-induced acute morphine tolerance may have NMDA receptor-dependent mechanisms.     

Environmental modification of tolerance to morphine discriminative stimulus properties in rats

The development of tolerance to the discriminative stimulus properties of morphine was examined in rats trained to discriminate saline and 3.2 mg/kg morphine under amultiple timeout 15 min, 5 min fixed-ratio 30 schedule of food delivery. Generalization gradients were generated by administering increasing doses of morphine before successive timeout periods within the experimental session. Over the course of the study, the minimal discriminable dose (MDD) of morphine under control conditions fluctuated but did not systematically increase or decrease. Acute pretreatments of 3.2–17.8 mg/kg morphine 4–24 h before a generalization test resulted in minor changes in the MDD. To examine development of tolerance, supplemental doses of morphine (17.8 mg/kg) or saline were administered twice daily while discrimination training was either suspended or continued. Tolerance was assessed by weekly generalization tests. Greater tolerance developed to the morphine stimulus when training was suspended than when training was continued. For both training conditions, response rates during generalization tests were markedly suppressed during supplemental morphine administration, and original generalization gradients were recaptured within 2 weeks after termination of supplemental morphine administration. Supplemental saline administration did not alter the discriminative or rate-altering effects of morphine under either training condition. Thus, the magnitude of tolerance to a morphine discriminative stimulus reflected an interaction of supplemental drug treatment with the training conditions imposed during that treatment.     

Generalization of [DAla2]-enkephalinamide but not of substance P to the morphine cue

Rats were trained to discriminate morphine (7.5 mg/kg, IP) from saline in a two bar positively reinforced lever pressing paradigm on a FR4 schedule. Morphine (IP) showed a naloxone reversible dose-related generalization to the training dose. [DAla²]-Methionine enkephalinamide (DAE) at 1 mg/kg and Substance P (SP) at 0.1 and 0.25 mg/kg showed vehicle appropriate responding after IP injection. DAE (5 mg/kg) disrupted responding completely; SP (0.5 and 0.1 mg/kg) disrupted responding in 50% of the rats. The disruption caused by IP injection of DAE was not naloxone reversible. Intraventricular injection of morphine (5 μg/rat) and DAE (5 μ/rat) produced generalization to the opiate cue. The effect of DAE was reversed by naloxone (1 mg/kg, SC). SP (500 and 750 ng/rat, IVT) produced saline-like responding; 1 μg/rat disrupted responding completely. These data demonstrate that morphine and enkephalin, but not Substance P, share similar discriminative properties.     

Peripheral receptors mediate the aversive conditioning effects of morphine in the rat

Previous evidence has shown that morphine produces positive reinforcing effects (as measured in the place conditioning paradigm) through an action in the central nervous system (CNS). The aversive conditioning effects of morphine (as measured in the place and taste conditioning paradigms) were produced when drug action was restricted to peripheral sites, particularly in the gut region. We now demonstrate that most of the aversive conditioning effects of morphine (using place and taste conditioning paradigms) are receptor mediated effects exerted through an action on peripheral opiate receptors. The conditioned taste aversions induced by intraperitoneal (IP) morphine (15 mg/kg) but not amphetamine (1 mg/kg) were attenuated by low IP doses of opiate antagonists (0.1 mg/kg of naltrexone or 1 mg/kg of the peripherally acting antagonist methynaltrexone (MN]. Morphine-, but not amphetamine-induced conditioned taste aversions were also attenuated in animals whose small sensory neurons, bearing the majority of primary afferent opiate receptors, were destroyed by neonatal treatments with capsaicin. In the place conditioning paradigm, the aversive conditioning effects produced by low IP administrations of morphine were blocked by opiate antagonists. Intraperitoneal pretreatments with 1 mg/kg of the quaternary opiate antagonist MN (which does not cross the blood-brain barrier effectively) were shown to block the conditioned place aversions produced by low IP doses of morphine (0.05 mg/kg), but not the place aversions produced by lithium chloride (75 mg/kg IP), or by high doses of naloxone (10 mg/kg SC). These results demonstrate that the aversive conditioning effects of morphine are primarily mediated through an action on peripheral opiate receptors.(ABSTRACT TRUNCATED AT 250 WORDS)     

Determination of cross tolerance in rat spinal cord using intrathecal infusion via sequential mini-osmotic pumps

Continuous intrathecal (IT) infusion via ALZET mini-osmotic pumps was used to induced spinal tolerance to morphine in the rat. Naloxone (1 mg/kg IP), injected on day 3 of continuous IT morphine (10 micrograms/hr), produced mild withdrawal symptoms in all morphine-treated animals. In rats pretreated with continuous IT morphine (10 micrograms/hr) or saline, systemic morphine (2, 4, 8, 10 and 15 mg/kg IP) produced equivalent, dose-dependent antinociception using the tail-flick and paw pressure tests. The rostral and caudal distribution of methylene blue dye in rat spinal cord was determined on days 1-7 of continuous IT infusion. The dye remained localized near the catheter tip throughout infusion; maximum distribution was 1.5 cm rostrally and 1.0 cm caudally. The data indicate that morphine, infused at the rate of 10 micrograms/hr, does not undergo extensive redistribution in the spinal cord. A sequential, double mini-osmotic pump technique for cross tolerance studies in rat spinal cord is described. In rats pretreated with continuous IT norepinephrine for 4 days, the antinociceptive actions of continuous IT morphine were reduced but not significantly different from saline-pretreated animals. These data suggest that morphine, injected into the spinal cord, does not produce behavioural analgesia by activation of local adrenergic systems.     

Inhibitory effects of MPEP,an mGluR5 antagonist,and memantine,an <Emphasis Type="Italic">N</Emphasis>-methyl-<Emphasis Type="SmallCaps">D</Emphasis>-aspartate receptor antagonist,on morphine antinociceptive tolerance in mice

Abstract Rationale. Inhibition of N-methyl-D-aspartate (NMDA) receptors by memantine, an NMDA-receptor antagonist, and other antagonists of ionotropic receptors for glutamate inhibit the development of opiate antinociceptive tolerance. The role of metabotropic receptors for glutamate (mGluR) in opiate tolerance is less known. Objective. In the present study, we examined the effect of 2-methyl-6-(phenylethynyl)-pyridine (MPEP), the mGluR type-I (subtype mGluR5) antagonist, as well as the effect of co-administration of low doses of memantine and MPEP on morphine antinociceptive tolerance in mice. Methods. Morphine antinociceptive activity was tested twice, before and after chronic morphine administration, in the tail-flick test using a cumulative dose–response protocol. Tolerance was induced by six consecutive days of b.i.d. administration of morphine (10 mg/kg, s.c.). Saline, memantine (7.5 mg/kg and 2.5 mg/kg, s.c.), MPEP (30 mg/kg and 10 mg/kg, i.p.) and the combination of both antagonists at low doses was given 30 min prior to each morphine injection during its chronic administration. A separate experiment assessed the effects of memantine, MPEP and their combination on acute morphine antinociception using a tail-flick test. Results. MPEP (30 mg/kg but not 10 mg/kg) as well as memantine (7.5 mg/kg but not 2.5 mg/kg) attenuated the development of tolerance to morphine-induced antinociception. When given together, the low doses of MPEP (10 mg/kg) and memantine (2.5 mg/kg) also significantly attenuated opiate tolerance. None of the treatments with glutamate antagonists produced antinociceptive effects or significantly affected morphine-induced antinociception. Conclusions. The data suggest that both mGluR5 and NMDA receptors may be involved in the development of morphine antinociceptive tolerance. Electronic Publication     

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.     

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.     

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.     

Environment-specific cross-sensitization between the locomotor activating effects of morphine and amphetamine

Groups of eight rats each were preexposed on four occasions to 10 or 20 mg/kg morphine sulfate, IP, either in activity boxes where activity was measured for two hours (COND, conditioning groups) or in their home cages (UNPAIRED groups). On alternate days these groups were administered saline in the other environment. Two groups of eight rats each served as CONTROL groups (one for each preexposure dose) and were administered saline in both environments. On the day following morphine preexposure, all animals were administered 0.5 mg/kg d-amphetamine sulfate, IP, prior to being tested in the activity boxes. On this test, the COND group preexposed to 10 mg/kg morphine showed higher levels of activity than either of its respective UNPAIRED or CONTROL groups. The COND group preexposed to 20 mg/kg morphine was significantly more active than its unpaired group, but not more active than its CONTROL group. The implications of such environment-specific cross-sensitization between the activity effects of opiate and stimulant drugs are discussed.     

Additive effect of dextromethorphan on the inhibitory effect of anti-NT4 on morphine tolerance

It has been proposed that opioid tolerance is a model of neuronal plasticity similar to learning and memory. Recent evidence suggests that neurotrophins may be involved in synaptic development and plasticity. Observations indicate that neurotrophin 4 (NT4) is required for the synaptic plasticity mediating both tolerance and memory. Also there are lines of evidence to indicate that NMDA receptors are involved in the neural plasticity underlying the development of opiate tolerance. Neurotrophins affect central transmission postsynaptically by enhancing NMDA receptor responsiveness. So we used the clinically available NMDA receptor antagonist, dextromethorphan, and the neurotrophin 4 antibody, anti-NT4, concomitantly and alone to investigate their effects on morphine tolerance. Tolerance was induced by injecting morphine (7 and 10 mg/kg i.p.) once per day for 4 days. Anti-NT4 (1 microg/rat i.c.v.) was administered 15 min before morphine. Results showed that chronic concomitant treatment of anti-NT4 with morphine in both doses inhibited the development of morphine tolerance. Also acute treatment of anti-NT4 significantly reversed the tolerance that was induced by morphine 7 mg/kg but failed to reverse the tolerance of morphine 10 mg/kg. Dextromethorphan in both doses (10 or 30 mg/kg) has an additive effect on the inhibitory effect of anti-NT4 on the reversal of morphine tolerance (7 mg/kg). These findings provide additional support for the hypothesis that NMDA receptor and NT4 may be involved in neural plasticity underlying opiate tolerance.     

Elevated startle during withdrawal from acute morphine: a model of opiate withdrawal and anxiety

Rationale An elevated startle response has been observed in humans and animals during withdrawal from multiple substances of abuse, a phenomenon thought to reflect the anxiogenic effects of withdrawal. Although anxiety is a common symptom of opiate withdrawal, few studies have examined the effects of morphine withdrawal on acoustic startle.Objective To develop a procedure for assessing opiate dependence through measurement of the startle reflex in rats.Methods The effects of opiate withdrawal on startle were evaluated using both spontaneous and naloxone-precipitated withdrawal from an acute dose of morphine. The ability of the treatment drugs clonidine and chlordiazepoxide to block withdrawal-induced increases in startle was also tested.Results Spontaneous withdrawal from an injection of morphine sulfate produced a significant increase in acoustic startle 2 h (3.2 mg/kg) or 4 h (10 mg/kg) after drug administration. Morphine withdrawal (10 mg/kg morphine sulfate) precipitated by the opiate antagonist naloxone (2.5 mg/kg) also produced a significant increase in startle magnitude. This elevation of startle was blocked by both clonidine (35 g/kg) and chlordiazepoxide (10 mg/kg).Conclusions These data demonstrate that both spontaneous and precipitated withdrawal from an acutely administered opiate produce anxiety-like effects on acoustic startle. This paradigm may be useful in the study of anxiety and the early mechanisms of drug dependence.     

Tolerance to antinociceptive effects of morphine without tolerance to its effects on schedule-controlled behavior

The development of tolerance to behavioral effects of morphine was investigated in rats that responded on a two-lever, multiple-trial, multiple differential-reinforcement-of-low-rate fixed-ratio (mult DRL FR) schedule of food presentation. Stable performances were maintained when sessions were conducted just twice per week. The effects of cumulative doses of morphine (1.0–8.0 mg/kg) or chlordiazepoxide (CDP; 4.0–32.0 mg/kg) were evaluated once per week; saline injections were given in the intervening sessions. The effects of saline and morphine on nociception were also evaluated in hot-plate tests conducted on the same subjects 15 min after selected operant sessions. Initially, morphine produced dose-related decreases in response rates and reinforcement rates in the DRL and FR components as well as significant increases in hot-plate response latencies. Following weekly administration of morphine (1.0–8.0 mg/kg) for 10 weeks, there was little or no tolerance to its effects on operant behavior. In contrast, complete tolerance developed to the antinociceptive effects of morphine. These results suggest that tolerance to various behavioral effects of morphine may be dissociated, and that the loss of reinforcement may be insufficient by itself to produce tolerance to effects of morphine on operant behavior. Additionally, whereas CDP initially produced only dose-related decreases in DRL and FR response rates, following weekly morphine the smaller doses of CDP (4.0–16.0 mg/kg) produced increases in response rates. Finally, the effects of cumulative doses of morphine did not differ significantly from the effects of noncumulative doses of the drug.     

The synergistic effect of concurrent spinal and supraspinal opiate agonisms is reduced by both nociceptive and morphine pretreatment

The antinociceptive effect of morphine administered into the periaqueductal gray (PAG), the intrathecal space (ITH) and concurrently, into both sites (in a 1:1 dose ratio), was assessed in 1) nontolerant rats, 2) rats made tolerant to the effect of morphine on the tail-flick (TF) test and 3) rats that were tested on the TF during chronic saline administration. In nontolerant rats, concurrent morphine injections produced a multiplicative antinociceptive effect (ED50 = 0.392 microgram, total dose) relative to that obtained after separate PAG (ED50 = 2.8 micrograms) or ITH (ED50 = 6.7 micrograms) injections. The multiplicative effect of concurrent morphine administration was significantly reduced in rats made tolerant to morphine (one 3 mg/kg SC injection and TF test per day for six days). Opiate synergy was also reduced but to a smaller extent in rats that were repeatedly tested on the TF during chronic saline administration (one SC injection and TF test per day for six days). Neither chronic morphine nor saline pretreatment altered the dose-response function to intrathecal morphine. However, both morphine and saline pretreatment significantly reduced the antinociceptive effect of morphine administered into the PAG. The data indicate that concurrent morphine administration into the PAG and ITH space results in a synergistic antinociceptive action which is reduced by performance of the nociceptive response, even in the absence of opiate administration. We suggest that the decrease in opiate synergism produced by nociceptive assessment (behavioral tolerance) is mediated supraspinally, while the additional decline resulting from morphine administered in conjunction with the nociceptive tests (opiate tolerance) is mediated by a combined action at spinal and supraspinal sites.     

Differences in morphine-induced antinociception in male and female offspring born of morphine exposed mothers

Objective:

Antinociceptive effect of morphine in offspring born of mothers that received saline or morphine during the gestation period was investigated.

Materials and Methods:

Wistar rats (200-250 g) received saline, morphine 0.5 mg/kg or 5 mg/kg during gestation days 14-16. All pups after weaning were isolated treatment/sex dependently and were allowed to fully mature. The antinociceptive effect of morphine was assessed in formalin test. Morphine (0.5-7.5 mg/kg) or saline (1 ml/kg) was injected intraperitoneally 10 min before formalin (50 μl of 2.5% solution in right hind-paw).

Results:

Male offspring born of saline-treated mothers were less morphine-sensitive than females. On the contrary, male offspring exposed prenatally to morphine (5 mg/kg) were more sensitive to morphine-induced antinociceptive response in formalin test. However, no difference in antinociceptive effect was observed amongst offspring of either sex born of mothers treated with morphine 0.5 mg/kg, identifying a lower dose effect of the opioid.

Conclusion:

The exposure to morphine during the developmental period may result in altered development of tolerance to morphine and thus involved in drug abuse.KEY WORDS: Formalin test, gestation, morphine, offspring, pain     

Influence of antineoplastic drugs on morphine analgesia and on morphine tolerance

The possible influence of cisplatin, methotrexate, adriamycin and vincristine on thermal pain threshold, morphine analgesia and development of morphine tolerance was investigated in mice. In the hot-plate test, the nociceptive threshold was not affected by acute or repeated administration of any of the antineoplastic drugs used. The analgesic activity of morphine was significantly reduced by pretreatment with cisplatin, intraperitoneally (i.p.) injected at the dose of 2 mg/kg. In contrast, methotrexate, subcutaneously (s.c.) injected at the dose of 1 and 5 mg/kg, adriamycin (1 and 3 mg/kg s.c.), vincristine (0.25 and 0.5 mg/kg i.p.) and a lower dose of cisplatin (1 mg/kg i.p.) had no effect. The development of tolerance to morphine analgesia was delayed by adriamycin but was not influenced by the other antineoplastic drugs used. These data show that, of the four antineoplastic agents used in this study, cisplatin may interfere in the mechanism of action of morphine, and that adriamycin may delay the development of opiate tolerance.     

Effects of post-ethanol administration of NMDA and non-NMDA receptor antagonists on the development of ethanol tolerance in C57BI mice

The effect of post-ethanol administration of NMDA and non-NMDA receptor antagonists on the development of environment-dependent ethanol tolerance was studied in C57B1 mice. Ethanol tolerance was produced by daily injections of ethanol (3.5 g/kg, IP) in the same experimental environment and measured as ethanol-produced sleep-time during 5 consecutive days. The non-competitive NMDA receptor antagonist, dizocilpine (MK-801; 0.1 mg/kg, IP), and the competitive NMDA receptor antagonist, CGP 39551 (5 mg/kg, IP), both given 120 min after the administration of ethanol, inhibited the development of tolerance to the hypnotic actions of ethanol. In contrast, the development of ethanol tolerance was not altered by administration of the specific AMPA/KA receptor blocking agents, NBQX (10 mg/kg, IP), and LY326325 (2.5 mg/kg, IP), respectively. Modulation of NMDA receptor activity by drugs like NMDA,d-cycloserine, and milacemide, which are known to enhance learning and memory in rodents, had no significant effect on the development of ethanol tolerance. Our present data confirm and extend previous findings which indicate that NMDA, but not non-NMDA, glutamate receptors may play an important role in the neuroadaptive processes associated with the development of ethanol tolerance.