Apparent absence of nicotine-induced conditioned place preference in rats

The conditioned place preference (CPP) paradigm was used in order to assess the reinforcing actions of nicotine in rats. Subjects were tested in unbiased two-compartment shuttle boxes, so-called because neither compartment was consistently preferred prior to drug conditioning. In the first experiment, subjects that were initially drug naive showed neither a preference nor an aversion to the compartment that had been paired on four occasions with injection of nicotine (0.2–0.8 mg/kg SC); a similar result occurred in another group given daily injections of nicotine in the home cage prior to the experiment. In a second experiment, nicotine (0.4, 0.8 mg/kg SC) again failed to produce a CPP, whereas marked CPPs were seen in parallel groups of rats tested with either d-amphetamine or methylphenidate. Although nicotine has been reported to produce conditioned place preference, the present results suggest that it is not a robust phenomenon.     

The effect of nicotine on the swimming speed of pre-trained rats through a water alley

Summary In sessions of ten runs each, swimming time of rats through a 4 m long water alley was measured. Four doses of nicotine (0.05; 0.1; 0.2; 0.4 mg/kg given intraperitoneally 30 minutes before testing) were tested in sessions with a braking load on the tails of the animals either in all 10 runs of a session, or in every second run, or in none of the 10 runs. Regardless of the swimming condition, nicotine produced a considerable, and at doses of 0.1 mg/kg and over, significant decrease of performance in the first two runs. From the third to the 10th run, the changes caused by nicotine were smaller and differed depending on the swimming conditions.A dose of 0.1 mg nicotine/kg improved performance in the without-load-sessions and the without-load-runs of the alternating sessions, while both 0.1 and 0.2 mg/kg improved performance of the with-load-runs of the alternating sessions. Performance in the without-load-sessions and the without-load-runs was depressed by 0.4 mg/kg and that in the with-load-sessions by 0.2 and 0.4 mg/kg.     

Nicotine conditions place preferences after intracerebral administration in rats

A single-trial place conditioning procedure, one treatment and one non-treatment during two daily conditioning sessions followed by a single test session on the 3rd day, was used to examine the place conditioning effects of intracerebrally administered nicotine. In the first series of experiments, Sprague-Dawley male rats were implanted unilaterally with guide cannulas aimed at the lateral ventricle. After 1 week, rats received either treatment (nicotine in 2 µl phosphate buffer or 2 µl of buffer alone) or no treatment (no injections) before being placed in the black or white compartment of a three-compartment place-conditioning apparatus for 20 min. The next day the rats received the opposite treatment before being conditioned in the opposite compartment. On day 3, animals had free access to the entire apparatus for 15 min and the time spent in each compartment was recorded automatically. Even though the rats exhibited a baseline bias for the black compartment, intracerebroventricular nicotine induced positive place preferences relative to buffer control, i.e. if treatments were paired with the black compartment, nicotine enhanced the preference for the black compartment, and if the treatments were paired with the white compartment, nicotine induced a preference for the white compartment. In addition, the nicotine-induced preference response was antagonized by the co-intraventricular administration of mecamylamine. In a second series of experiments, animals were implanted unilaterally with guide cannulas aimed at the pendunculopontine tegmental nucleus of the mesopontine tegmentum. Nicotine microinjection, 1.2–18.5 nmol in 0.5 µl buffer, induced a dose-dependent positive place preference response. The place preference induced by 18.5 nmol nicotine was antagonized by co-administering 0.8 nmol mecamylamine; 0.8 nmol mecamylamine had no effects of its own. Finally, microinjections of 18.5 nmol nicotine 2 mm dorsal to the pedunculopontine tegmental nucleus did not condition place preferences. These data demonstrate that a sensitive site of action of the nicotine-conditioned place preference response in rats is within the mesopontine tegmentum.     

Effects of naloxone,metenkephalin, and morphine on phencyclidine-induced behavior in the rat

The effects of naloxone, metenkephalin, and morphine were tested on phencyclidine(PCP)-induced stereotyped behaviors, ataxia, and hyperactivity in the rat. Naloxone (8 mg/kg) significantly decreased stereotypy, ataxia, and hyperactivity across all PCP doses tested (2.0, 4.0, and 6.0 mg/kg). Metenkephalin (40 g/kg) and morphine (5 and 10 mg/kg) increased ataxia at the 4.0 and 6.0 mg/kg PCP doses. Stereotypy was altered by the opiates in a dose-dependent manner; enhanced by metenkephalin (40 g/kg) at 2.0 mg/kg and inhibited by metenkephalin (40 g/kg) and morphine (10 mg/kg) at 4.0 and 6.0 mg/kg PCP. Locomotor activity was increased by morphine (5 mg/kg) at 2 mg/kg PCP. These results suggest an involvement of central opiate receptor mechanisms in the mediation of PCP-induced behaviors in the rat.     

Evidence for increased apomorphine-sensitive dopaminergic effects after acute treatment with morphine

Apomorphine in a very small non-stimulant dose 0.05 mg/kg s.c. antagonized the locomotor stimulant effect of morphine (50 mg/kg) in mice. Apomorphine (0.05 mg/kg) antagonized also the locomotor stimulant effect of a single low dose of morphine (2 mg/kg) in the rat as well as the late stimulatory effect seen after a higher dose of morphine (10 or 20 mg/kg). The pretreatment with morphine in increasing doses induced an increasing degree of potentiation of stereotyped behavior induced by high doses of apomorphine (0.5 and 2 mg/kg s.c.) in the rat. The apomorphine stereotyped biting/gnawing activity was found strongly increased by morphine (10–50 mg/kg) during the time interval where morphine given alone induced catalepsy. Our results indicate that the acute treatment with morphine may induce an increase of apomorphine sensitive dopaminergic mechanisms. The inhibitory effect by a very small dose of apomorphine (0.05 mg/kg) of morphine stimulation may be due to a presynaptic dopamine receptor stimulant effect of apomorphine. The antagonism by the serotonin antagonist methergoline of morphine catalepsy and the increase by this drug of the late morphine excitation indicate in addition a role of a serotonergic mechanism.     

Facilitation of memory retrieval by pretest morphine mediated by μ but not δ and κ opioid receptors

Mice were trained to avoid electric shock (0.6 mA) in a step-through type passive avoidance learning task, retention being measured 24 h after the training trial. Morphine 10 mg/kg administered 30 min before the test trial (pretest) facilitated memory retrieval, and the effect was completely antagonized by 1 mg/kg naloxone, a selective -opioid receptor antagonist. On the other hand, pretest administration of 0.01–10 mg/kg DTLET, a selective -opioid receptor agonist, did not produce the same effect as morphine. Nor-binaltorphimine, a -opioid receptor antagonist, did not antagonize the effect of pretest morphine, at doses of 1 and 2 mg/kg. These results suggest that the facilitation of memory retrieval by pretest morphine is mediated through - but not - or -opioid receptors.     

Evidence that the aversive effects of opioid antagonists and κ-agonists are centrally mediated

The role of central versus peripheral opioid receptors in mediating the aversive effects of opioids was examined by use of an unbiased place preference conditioning procedure in rats. The non-selective opioid antagonist naloxone (NLX) produced conditioned aversions for the drug-associated place after subcutaneous (SC) as well as intracerebroventricular (ICV) administration. Place aversions were also observed in response to the ICV administration of the selective -antagonist CTOP. In contrast, the selective -antagonist ICI 174,864 and the selective -antagonist norbinaltorphimine (nor-BNI) (ICV) were without effect. Place aversions were also produced by central applications of the selective -agonist U50,488H and the dynorphin derivative E-2078. For those opioid ligands tested, the doses required to produce place aversions were substantially lower following ICV as compared to SC administration. These data confirm that -agonists and opioid antagonists produce aversive states in the drug-naive animal and demonstrate that this effect is centrally mediated. Furthermore, the ability of NLX and CTOP, in contrast to both ICI 174,864 and nor-BNI, to produce place aversions suggests that the aversive effects of opioid antagonists result from the blockade of -receptors.     

β-adrenergic antagonists attenuate withdrawal anxiety in cocaine-and morphine-dependent rats

Rats were treated chronically with either cocaine (20 mg/kg/day, 14 days), morphine (incrementing doses of 10 mg/kg/day to 80 mg/kg, 11 days) or saline. During morphine or cocaine abstinence (48 h), dependent rats showed increased anxiety-like behavior in a conditioned defensive burying paradigm as evidenced by significantly shorter latencies to begin burying as well as a 4-fold increase in burying duration relative to salinetreated animals. This withdrawal-induced increase in burying behavior was blocked by pretreatment with either the -adrenergic antagonist propranolol (5 mg/kg) or the lipophobic selective ₁-antagonist, atenolol (5 mg/kg). These results are consistent with the possibility that activation of peripheral ₁ receptors may substantially contribute to withdrawal-induced anxiety and that -adrenergic antagonists could be useful in treating in cocaine and morphine dependent addicts.     

Infusion of γ2-MSH produce a conditioned taste aversion in morphine-dependent rats

A two flavour, unbiased, taste preference conditioning procedure was used to test for possible motivating effects of ₂-MSH. Three training trials failed to produce any significant effect with doses ranging from 2.4 to 40 g/ ICV infusion in drug-naive, non-operated or placebo-implanted rats. However, in rats made dependent by SC implantation of a morphine pellet 4 days earlier 15 g ₂-MSH/infusion produced a taste aversion that was comparable to that produced by infusion of a low dose of the competitive opioid receptor antagonist naloxone (0.32 g). The findings confirm with a conditioning procedure and with opiate-dependent animals the naloxone-like effects of ₂-MSH. They also suggest that this endogenously-located peptide may acquire an aversive property as a result of chronic morphine treatment.     

Pharmacokinetic Drug–Drug Interaction of the Novel Anticancer Agent E7070 and Acenocoumarol

E7070 is a novel sulfonamide anticancer agent that arrests cancer cells at the G1/S boundary of the cell cycle. Three patients receiving chronic therapy with the oral anticoagulant acenocoumarol experienced bleeding and/or a prolonged prothrombin time after treatment with E7070 at a dose of 700mg/m² given as a 1-h infusion. In vitro studies have shown that E7070 has the potential to inhibit several cytochrome P450 (CYP)-enzymes, including CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4. The major enzyme involved in the metabolism of acenocoumarol in man is CYP2C9. This study was performed to investigate the interaction between E7070 and acenocoumarol.Blood samples were obtained from two patients receiving daily oral maintenance treatment with acenocoumarol both prior to and following treatment with E7070. In addition, we incubated acenocoumarol enantiomers with pooled human microsomes with and without E7070 and measured the in vitro plasma protein binding of acenocoumarol after incubation with E7070. Pharmacokinetic parameters of acenocoumarol were calculated by noncompartmental analysis and revealed that in both patients the area under the concentration–time curve up to 24h after the acenocoumarol administration was higher following E7070 (2.56 and 1.58h*mol/L) compared to the systemic exposure in the absence of E7070 (1.87 and 1.23h*mol/l). The formation of acenocoumarol metabolites was retarded by E7070 at already low concentrations (2.1M). The plasma protein binding of acenocoumarol was reduced at higher concentrations of E7070 (259M).These results indicate that E7070 may primarily interact with acenocoumarol by reducing its systemic clearance. Displacement of acenocoumarol's plasma protein binding by E7070 may also occur but to a minor extent. In the absence of careful monitoring this drug–drug interaction may result in hypoprothrombinemia and a hemorrhagic tendency.     

Nicotine-induced place preferences following prior nicotine exposure in rats

The motivational properties of morphine and nicotine were investigated in an automated conditioned place preference (CPP) procedure using a two-compartment apparatus. The accuracy of the photocell recording system was assessed by correlation with direct observation. In a counterbalanced conditioning design, graded doses of morphine (0.1–3.2 mg/kg SC) produced dose-related CPP. Under similar conditions, a dose of nicotine (0.6 mg/kg SC) previously reported to produce CPP failed to show an effect. Increasing the number of conditioning trials from 4 to 12 did not facilitate CPP with nicotine. After pretreatment with nicotine (0.4 mg/kg SC) daily for 7 days prior to conditioning, nicotine (0.4–0.8 mg/kg) produced increasing magnitudes of CPP. Locomotor activity was assessed during both conditioning and extinction tests. During conditioning, nicotine but not morphine decreased activity in the first conditioning trial, but by the fourth trial, marked stimulation was apparent following administration of either drug. Activity in the drug-paired compartment was not increased during tests for CPP carried out in the undrugged state following 4 conditioning trials with either morphine or nicotine, but there was evidence for conditioned hyperactivity after 12 conditioning trials with nicotine. The results suggest that motivational properties of nicotine can be detected in counterbalanced CPP procedures, but only in subjects with a history of nicotine exposure. The CPP produced by morphine or nicotine does not appear to be an artefact associated with conditioned changes in locomotor activity.     

The role of descending inhibition in the antinociceptive effects of the pyrazolone derivatives,metamizol (dipyrone) and aminophenazone (“Pyramidon”)

Summary The study was carried out to provide further evidence that the two pyrazolone derivatives, metamizol and aminophenazone, produce central antinociceptive effects by stimulating inhibition descending from the periaqueductal grey (PAG) to the spinal cord. Experiments were carried out on rats in which the tail-flick response to radiant heat, nociceptive activity in ascending axons of the spinal cord, and activity of neurones in the PAG and the substantia nigra were studied. Microinjection of procaine (10 g) into the PAG reduced the tail-flick latency and abolished the increase in latency caused by i.p. injection of metamizol (40 mg/kg) and aminophenazone (150 mg/kg); it did not significantly reduce the antinociceptive effect of i.p. injection of morphine (2 mg/kg). Threshold doses of morphine (1 and 2 g) administered by intrathecal (i.t.) injection potentiated the effect of threshold doses of metamizol injected i.p. (10 mg/ kg) or into the PAG (10 g) in the tail-flick test. Morphine (2 g) injected i.t. potentiated the effect of i.v. injection of metamizol (80 mg/kg) on nociceptive activity in ascending axons by eliminating the stimulant effect of metamizol on about one third of the axons. Threshold doses of morphine injected i.t. failed to potentiate the antinociceptive effect of aminophenazone (50 mg/kg) injected i.p. in the tail-flick test. The results support the view that metamizol and aminophenazone activate pathways descending from the PAG and exerting an inhibitory effect on nociceptive impulse transmission at the spinal level.Supported by the Schwerpunkt Nociception and Schmerz of the Deutsche Forschungsgemeinschaft Send offprint requests to I. Jurna at the above address     

Anti-nociceptive effect of morphine,opioid analgesics and haloperidol injected into the caudate nucleus of the rat

Summary The effect of intracaudate (i.c.) microinjections of morphine, opioid analgesics and haloperidol was determined on the tail-flick response evoked by radiant heat in rats. Bilateral injections (0.2 l on each side) into the caudate nuclei of morphine 5 g, pethidine 50 g, levorphanol 4 g, dextrorphan 10 g and haloperidol 5 g significantly increased the reaction time of the tail-flick response. The antinociceptive effect of an i.c. injection of morphine or levorphanol was abolished by an intraperitoneal (i.p.) injection of naloxone 0.2 mg/kg or apomorphine 2 mg/kg. The anti-nociceptive effect of pethidine, dextrorphan and haloperidol was reduced but not abolished by an i.p. injection of naloxone 0.2 mg/kg. An i.p. injection of apomorphine 2 mg/kg abolished the effect of an i.c. injection of haloperidol. A bilateral i.c. injection of naloxone 5 g or apomorphine 10 g reduced the anti-nociceptive effect of an i.p. injection of morphine 2 mg/kg or haloperidol 2 mg/kg, but did not abolish it. It is concluded that (1) an anti-nociceptive effect can be achieved by an action on the caudate nucleus of the drugs tested; (2) the anti-nociceptive effect exerted by morphine and levorphanol in the caudate nucleus is due to a specific action mediated by opiate receptors, whilst that produced by pethidine and dextrorphan is due to a specific and/or unspecific action; (3) the anti-nociceptive effect of haloperidol in the caudate nucleus is due to an impairment of dopaminergic impulse transmission, which is also involved in the effect of morphine and levorphanol.Supported by the Sonderforschungsbereich 38 Membranforschung     

Rewarding properties of β-endorphin as measured by conditioned place preference

The role of -endorphin as a possible mediator in the reinforcing properties of opiates was investigated using a conditioned place preference paradigm. Heroin, a synthetic opiate known to have reinforcing properties, produced a strong preference for an environment previously paired with heroin injection at all doses tested (0.25, 0.5, 1.0, 2.0 mg/kg SC). No such place preference was observed following saline injections. Rats also showed dose-dependent place preference for the environment paired with -endorphin when injected intracerebroventricularly (significant dose was 2.5 g). At higher doses (5.0 and 10.0 g) rats showed no preference for the paired environment, but were catatonic. Pretreatment with naloxone (0.04, 0.2, 1.0 mg/kg SC) attenuated the rewarding effect of -endorphin (2.5 g) at all doses tested. The lowest dose of naloxone which had no aversive effect when tested alone could also significantly block the positive effect of -endorphin. The reinforcing dose of -endorphin (2.5 g) also produced an increase in locomotor activity, when tested in photocell cages. This suggests that the hyperactivity induced by -endorphin may contribute to the preference for an environment previously paired with the same drug. The reinforcing effect of -endorphin is most probably mediated by the mu and/or delta opioid subtype receptor, since -endorphin has a high affinity for these receptors. These results demonstrate positive reinforcing properties of -endorphin in the central nervous system.     

Tolerance to the behavioral and neurochemical effects of haloperidol and morphine in rats chronically treated with morphine or haloperidol

Summary The acute administration of morphine sulfate (79 moles/kg) or haloperidol (6.65 moles/kg) produced catalepsy and concomitant increase in striatal dopamine turnover in rats. The animals made dependent on morphine by 52 morphine injections (maintenance dose of 1056 moles/kg/day, given in four daily doses) and then tested during 3 days of withdrawal from morphine, showed tolerance to the cataleptic and the neurochemical effects of morphine as well as those of haloperidol. That tolerance was not seen after 14 days of withdrawal from morphine. The animals chronically treated with haloperidol for 12 days (maintenance dose of 53.2 moles/kg/day, given in two daily doses) and then tested 72 h after last haloperidol injection, did not show tolerance to the cataleptic or the neurochemical effect of haloperidol or morphine. These results suggest that dopaminergic systems underlying motor coordination and regulation of the neurotransmitter synthesis are among those susceptible to narcotic action and to the process of tolerance development during aarcotic dependence.     

Effects of morphine on release of acetylcholine in the rat striatum: an in vivo microdialysis study

Summary We examined the effect of morphine on the release of acetylcholine (ACh) in the striatum of freely moving rats using the in vivo microdialysis method. The basal level of ACh was 3.01 ± 0.51 pmol/30 l/15 min in the presence of neostigmine (10 M). Tetrodotoxin (1 M), a selective blocker of voltage-dependent Na⁺ channels, rapidly decreased the release of ACh in the striatal perfusates. Morphine at a dose of 10 mg/kg (i.p.) caused a reduction of ACh release in the striatum at 90–150 min. However, a lower dose of morphine (5 mg/kg, i.p.) did not affect ACh release in the striatum. The reduction following intraperitoneal administration of morphine was abolished by naloxone (1.0 mg/kg).After microinjection of the neurotoxin 6-hydroxydopamine (6 g/3 l, 7 days before) in the substantia nigra, the morphine (10 mg/kg)-induced decrease of ACh was attenuated, and a similar result occurred following reserpine (2 mg/kg, i.p.) 24 h before combined with -methyl-p-tyrosine (300 mg/kg, i. p.) 2.5 h before.These findings indicate that morphine exerts an inhibitory influence on striatal ACh release in freely moving rats and that this inhibitory effect is mediated by the nigro-striatal dopaminergic system.Correspondence to K. Taguchi at the above address     

Conditioned taste aversions produced by nicotine in Roman High and Low Avoidance strains of rats

Rats of the RHA/iop and RLA/iop strains have been compared in a conditioned taste aversion procedure using nicotine (0.4 mg/kg SC) as the UCS. The procedure utilised a balanced, within-subject design for assessing discriminative aversions to drug- and saline-paired flavoured solutions. Nicotine produced clear aversions in both strains and there were no detectable differences in acquisition. During extinction, rats of the RHA/iop strain consumed more of the drug-paired flavoured solution than rats of the RLA/iop strain, and this difference became greater as the number of extinction trials proceeded. Differences in total fluid intake were too small to account for these effects that were also shown by changes in proportional intake when both flavoured solutions were presented simultaneously. Aversion was, therefore, rather weaker in RHA/iop rats than in RLA/iop rats. These results suggest that rats of the two strains do not differ in learning ability in a general way, and support interpretations based on differences in emotionality.     

Midazolam cue in rats: Effects of Ro 15-1788 and picrotoxin

The discriminative stimulus effect of midazolam, a short-acting benzodiazepine, was used for testing the effects of drugs thought to act as antagonists at different sites in the proposed benzodiazepine receptor complex. Rats were trained in a standard two-bar operant conditioning procedure with food reinforcers delivered on a tandem schedule. The 0.4 mg/kg dose of midazolam used for training was well discriminated, typically yielding at least 95% correct responding. The benzodiazepine receptor antagonist Ro 15-1788 blocked the discriminative effect of midazolam but did not influence generalization to pentobarbitone (7.5 mg/kg). The indirect GABA antagonist picrotoxin attenuated both generalization to pentobarbitone and its response rate-reducing effect. Picrotoxin had no effect on the discriminative effect of midazolam at 0.4 mg/kg but it blocked the effect of 01 mg/kg. Even in doses which reduced overall response rates, nicotine did not block discrimination of midazolam (0.4 mg/kg). The results are consistent with models which postulate a GABA-linked ion channel which is a site of action for barbiturates and which is downstream of the benzodiazepine receptor itself.     

Strain differences in the acquisition of nicotine-induced conditioned taste aversion

Lewis (LEW) and Fischer (F344) rat strains differ on a variety of physiological and behavioral endpoints, including reactivity to drugs of abuse. Although they differ in drug reactivity, such assessments are generally limited to morphine and cocaine. To determine if these differences generalize to other drugs, the present study examined these strains for their reactivity to the affective properties of nicotine, specifically their sensitivity to nicotine in the conditioned taste aversion preparation. For four or five conditioning cycles given every other day, rats from both strains were allowed access to saccharin and injected with nicotine (0.1, 0.4, 0.8 mg/kg) or vehicle. On intervening days, all rats were given access to water and injected with vehicle. Under this one-bottle training and testing procedure, neither strain displayed aversions at the lowest dose of nicotine (0.1 mg/kg). Aversions were evident for both strains at 0.4 and 0.8 mg/kg, although the F344 rats acquired the aversions at 0.4 mg/kg faster and displayed a significantly greater aversion at 0.8 mg/kg than subjects from the LEW strain. For both strains, aversions were evident at all doses (and in a dose-dependent manner) when subjects were given access to saccharin and water in a two-bottle test. There were, however, no strain differences on this test. Differences between the two strains in their acquisition of nicotine-induced taste aversions were discussed in the context of aversion assessments with other compounds as well as in relation to differences in the self-administration of nicotine in the two strains.     

Tolerance to nicotine in mice lacking α7 nicotinic receptors

Rationale Previous studies have suggested that a knockout of the gene coding for 7 nicotinic receptor subunits influences the behaviour of undrugged mice but not the acute effect of nicotine on locomotor activity.Objectives The present studies extend these observations to nicotine tolerance assessed by means of schedule-controlled behaviour.Methods Groups of 7^–/– and 7^+/+ mice were trained to press levers under an FR20 schedule of food reinforcement. The acute response rate-depressant effects of nicotine were determined in both genotypes and the mice were then subdivided into groups treated daily with nicotine (1.2 mg/kg/day) or saline. After 39 days of exposure to this regimen, the dose–response curves were re-determined.Results Knockout of the 7 gene had no consistent effect on the lever-pressing behaviour of undrugged mice and did not influence the acute, dose-related, response rate-depressant effect of nicotine (0.2–1.2 mg/kg). When dose–response curves for nicotine (0.4–2.0 mg/kg) were re-determined after daily dosing with the drug, both wild-type and knockout mice developed similar tolerance to nicotine, as shown by 2.5-fold shifts to the right of the dose–response curves.Conclusions Nicotinic receptors containing the 7 subunit do not play a significant role in the regulation of the lever-pressing behaviour studied or in the acute behavioural depressant effect of nicotine and the development of tolerance to that effect. Such results contrast with previous reports suggesting profound impairments in sensitivity to nicotine in nicotinic receptor 2^–/– mice.C. Naylor and D. Quarta are listed alphabetically and contributed equally to the work.