Influence of training history on ethanol discrimination in rats

The compound stimulus hypothesis of ethanol discrimination predicts that a history of training to discriminate drugs that mimic individual elements of the ethanol stimulus should attenuate stimulus control by other stimulus elements (associative blocking). Rats were trained initially to discriminate either chlordiazepoxide (5 mg / kg s.c., n = 10) or dizocilpine (0.08 mg / kg i.p., n = 10) from vehicle in two-lever procedures with food reinforcers presented on a tandem variable-interval fixed ratio schedule. Control rats received 'sham training' (vehicle injections only, n = 9). All subjects were then trained to discriminate ethanol (1.5 g / kg intragastrically (i.g.)) until discrimination accuracy reached 95%. Chlordiazepoxide (1.25-10.0 mg / kg s.c.) produced more drug-appropriate responding in rats with a previous history of training to discriminate chlordiazepoxide than in either of the other two groups, but stimulus control by dizocilpine was not attenuated. Equivalent results were obtained in rats with a previous history of training to discriminate dizocilpine. Ethanol (0.375-3.0 g / kg i.g.) produced similar dose-related increases in drug-appropriate responding in all three groups. Thus, previous discrimination training modified the characteristics of ethanol discrimination in a way that may be explained by persistence of the original discriminations. The lack of evidence for associative blocking contrasts with results of previous experiments on the discrimination of compound stimuli produced by administering drug mixtures. The findings provide limited support for the hypothesis that ethanol produces a compound stimulus that includes elements of positive modulation of gamma-aminobutyric acid (GABA(A)) receptors and of N-methyl-D-aspartate (NMDA) antagonism.     

Influence of training paradigm on specificity of drug mixture discriminations.

Generalization to different drugs and drug mixtures has been examined in rats trained to discriminate a mixture of amphetamine (0.4 mg/kg) plus pentobarbitone (10 mg/kg) from saline (AND discrimination, n = 8) or to discriminate the same mixture from its component drugs alone (AND-OR discrimination, n = 9). The studies used two-lever operant procedures with a tandem variable interval 1-min fixed-ratio 10 schedule of food reinforcement. There was partial generalization to nicotine and midazolam and no generalization to cocaine, caffeine, or ethanol under AND-discrimination conditions and no generalization to any of these drugs in the AND-OR discrimination. Nicotine or midazolam coadministered with the training doses of pentobarbitone and amphetamine, respectively, produced full generalization in the AND discrimination and partial generalization under AND-OR conditions. Cocaine coadministered with pentobarbitone generalized fully under both procedures, but at larger doses in the AND-OR than in the AND discrimination. Mixtures of either nicotine plus midazolam or caffeine plus ethanol produced very marked generalization under AND-discrimination conditions, but were without significant effect in the AND-OR procedure. The results consistently supported the hypothesis that the AND-OR discrimination procedure increases the specificity of discriminations based on drug mixtures.     

Influencing the specificity of drug mixture discriminations by varying the training procedure

Studies of the discriminative stimulus effects of drug mixtures provide an approach to polydrug abuse and to studies on single drugs with multiple effects. The experiments described here investigated whether the use of the AND-OR procedure increases the specificity of drug mixture discriminations. Rats were trained to discriminate a mixture of nicotine (0.4 mg/ kg) plus midazolam (0.2 mg/kg) from saline (AND-discrimination, n = 10) or to discriminate the same mixture from its component drugs alone (AND-OR discrimination, n = 10). The studies used two-lever operant procedures with a tandem variable interval 1 min fixed ratio 10 (FR 10) schedule of food reinforcement. Under AND-discrimination conditions, there was partial generalization to amphetamine and pentobarbitone when each drug was administered singly. With the AND-OR-discrimination, there was no generalization to amphetamine and partial generalization to pentobarbitone. In 'single substitution' tests, pentobarbitone or amphetamine was co-administered with the training doses of nicotine and midazolam, respectively; there was full generalization in the AND-discrimination and no generalization under AND-OR conditions. In 'dual substitution' tests, mixtures of amphetamine plus pentobarbitone produced full generalization under AND-discrimination conditions, and partial generalization in the AND-OR procedure. Wherever comparisons were made, generalization was less under AND-OR- than under the AND-discrimination procedure, confirming that the AND-OR procedure can increase the specificity of discriminations based on drug mixtures. The similarity with findings reported previously for training with mixtures of amphetamine plus pentobarbitone suggests that this may reflect a general principle rather than a phenomenon restricted to particular training drugs.     

AND and AND-OR drug mixture discriminations in rats: generalization to single drugs and drug mixtures

Rationale: Studies of the discriminative stimulus effects of drug mixtures provide an approach to polydrug abuse and studies on single drugs with multiple effects. Objective: This study was designed to investigate whether the use of the AND-OR procedure increases the specificity of drug mixture discriminations. Methods: Rats were trained to discriminate a mixture of amphetamine (0.4 mg/kg) plus pentobarbitone (10 mg/kg) from saline (AND-discrimination, n = 8) or to discriminate the same mixture from its component drugs alone (AND–OR discrimination, n = 9). The studies used two-lever operant procedures with a tandem variable interval 1-min fixed ratio 10 schedule of food reinforcement. Results: Under AND-discrimination conditions, there was partial generalization to nicotine and midazolam when each drug was administered singly, and there was no generalization to cocaine, caffeine or ethanol. With the AND-OR discrimination, there was no generalization to any of the preceding drugs administered singly. In “single substitution” tests, nicotine or midazolam was co-administered with the training doses of pentobarbitone and amphetamine, respectively; there was full generalization in the AND-discrimination and partial generalization under AND-OR conditions. Cocaine co-administered with pentobarbitone generalized fully under both procedures, but the dose of cocaine needed was much larger in the AND-OR than in the AND-discrimination. In “dual substitution” tests, mixtures of two novel substances were tested. Mixtures of either nicotine plus midazolam or caffeine plus ethanol produced very marked generalization under AND-discrimination conditions, but were without significant effect in the AND-OR procedure. Throughout the studies, in every instance where comparisons were made, generalization was greater or occurred at lower doses under AND- than under the AND-OR discrimination. Conclusions: The study yielded extensive evidence supporting the hypothesis that the AND-OR discrimination procedure increases the specificity of discriminations based on drug mixtures. Received: 6 May 1998/Final version: 4 October 1998     

Discriminative stimulus functions of CNS sedative drugs assessed by drug versus drug discrimination procedures in gerbils

 This study is concerned with dissecting out differences in the discriminative stimulus attributes between drugs from broader pharmacological categories such as sedative/hypnotics where generalization tests often indicate shared stimulus effects. To this end, the discriminative stimulus effects of three to five doses of either chlordiazepoxide (CDP), chlormethiazole (CMZ), ethanol (ETOH), and pentobarbital (P-barb) were studied with gerbils in a two-choice, T-maze task. When the discrimination was based upon the presence versus the absence of drug administration, speed of acquiring the discrimination increased dose-dependently for all four drugs with log of sessions to criterion being a linear function of log dose. The slopes of all four lines were similar. The acquisition of a discrimination based on different doses of the training drug was then examined. A two-to-one ratio of high to low doses was used. Discriminative control developed with CMZ, ETOH, and P-barb, but not with CDP. The side of the T-maze that correlated with the lower training dose was always selected in tests with saline. Gerbils were then trained to discriminate between two drugs using at least two different dose combinations. The CDP versus CMZ, CDP versus ETOH, CMZ versus P-barb, CMZ versus ETOH discriminations were acquired. The CDP versus P-barb discrimination was obtained across doses only after an additional training procedure was instituted such that the P-barb dose was incremented gradually from low to higher doses during discrimination training; the CDP training doses were 20 and 30 mg/kg in two different groups of gerbils, respectively and the dose of P-barb was incremented by 2.5 mg/kg until the final dose of 20 mg/kg was reached with the barbiturate. Thus, although often shown to share similar stimulus effects, members of the broad pharmacological class of CNS sedatives/hypnotics nonetheless were shown to be discriminable from one another. Non-drug tests (vehicle) were used to assess stimulus control between pairs of drugs, equal control being implicated when responding was evenly distributed across the two training conditions. Received: 28 May 1997 / Final version: 11 July 1997     

Training dose as a decisive factor for discrimination of a drug mixture in rats

The impact of training dose on the characteristics of a discrimination maintained by a mixture of two dissimilar drugs has been investigated in order to refine this approach to the study of drug interactions. Three groups of rats (n = 10) were trained to discriminate mixtures of (+)-amphetamine (0.2-0.8mg/kg) plus pentobarbitone (5-20mg/kg) from saline, in a two-lever operant procedure with food reinforcement, with the ratio of the doses held constant (amphetamine: pentobarbitone, 1:25). Discriminations were acquired to an accuracy of 90-97%. There was full generalisation to amphetamine alone, but only in rats trained with mixtures of the smaller doses of the single drugs. There was partial generalisation when either apomorphine (50%) or nicotine (63%) was administered alone, and the magnitude of these responses was inversely related to the dose of mixture used for training. Doses of pentobarbitone half of those used for training produced little discriminative response when administered alone to rats trained with the two smallest doses of the mixture; the same doses of pentobarbitone increased responses to amphetamine or apomorphine in a more than additive manner. Strikingly, some doses of apomorphine and pentobarbitone that did not generalise when administered separately, produced full generalisation when administered together, but only in rats trained with the smaller doses of the mixture. In contrast, pentobarbitone did not enhance generalisation to nicotine in any group. It was concluded that, on the one hand, patterns of generalisation to single drugs followed an orderly pattern resembling those for discriminations established with single drugs. On the other hand, there was a complex pattern of generalization from one mixture to another; thus, altering the doses of drugs used for training markedly influenced discriminations of an abused drug mixture, but no simple rules to predict the influence of training dose have been ascertained.     

The chlordiazepoxide/pentylenetetrazol discrimination: characterization of drug interactions and homeostatic responses to drug challenges

Rats were trained to discriminate chlordiazepoxide (CDP) from pentylenetetrazol (PTZ) in a two-lever food motivated discrimination task. Training drug doses were adjusted until subjects emitted approximately 50% of their responses on each of the two drug-appropriate levers during saline injection tests. Tests that followed injection of CDP/PTZ combinations illustrated a reciprocal antagonism between the two drugs. Saline-injection tests that followed large dose injections of CDP revealed a period of predominantly PTZ-appropriate responding that persisted after the initial period of predominantly CDP-appropriate responding. These data are interpreted to suggest that, unlike some other drugs that have been shown to antagonize the behavioral and CNS effects of benzodiazepines, the interoceptive stimulus generated by PTZ occupies a position opposite to that of CDP along some single affective continuum. In addition, these data suggest that drug/drug (DD) discriminations are capable of characterizing the interactions between training drugs. Finally, the data suggest that the CDP/PTZ discrimination is a sensitive detector of bidirectional shifts in interoceptive stimulus state along the CDP/PTZ continuum.     

Behavioral interactions caused by combined administration of morphine and MK-801 in rats

Rationale: The NMDA antagonist MK-801 reportedly blocks experience-dependent changes in sensitivity to morphine, including tolerance to its analgesic actions and sensitization to its locomotor-stimulating effects. However, evidence in the existing literature suggests that some of MK-801’s effects are additive (or synergistic) with those of morphine. Objectives: Experiments were conducted to characterize the effects of acute and repeated administration of the combination of MK-801 and morphine on analgesia, locomotor activity, and drug discrimination in rats. Methods: In each experiment, rats were first tested repeatedly after treatment with the combination of MK-801 and morphine, and then after treatment with either drug alone. Results: The analgesic effects of MK-801 combined with morphine were greater than those of morphine alone, but tolerance to the combination of drugs developed at a similar rate as to morphine alone. The locomotor-stimulating effects of MK-801 combined with morphine were also greater than those of either drug alone, and locomotor sensitization developed to the combination of drugs but not to either drug alone at the low doses used. Rats learned to discriminate a combination of MK-801 and morphine from vehicle as quickly as they learned to discriminate morphine alone from vehicle, but those trained with the combination of MK-801 and morphine responded primarily at the vehicle-appropriate lever when given either drug alone. Conclusions: Since behavioral adaptations readily occur in the presence of MK-801, it appears that NMDA antagonists fail to invariably block the cellular plasticity that underlies such adaptations. Rather, the expression of adaptations in drug sensitivity appears related, at least in part, to the continued presence of the discriminative stimulus cues that are present during conditioning. Although NMDA receptors are important for some forms of cellular plasticity, the present studies illustrate the difficulty in interpreting behavioral studies in which MK-801 is given with morphine. Received: 24 November 1999 / Accepted: 25 March 2000     

Ethanol effects in pigeons trained to discriminate MK-801, PCP or CGS-19755

The non-competitive N-methyl-D-aspartate (NMDA) antagonists MK-801, PCP and ketamine have recently been found to produce full drug-appropriate responding in pigeons trained to ethanol (1.5g/kg) in a two-key operant drug discrimination procedure. In the present study, ethanol (0.56-3.2g/kg i.g.) was administered to pigeons trained to discriminate MK-801 (0.18mg/kg, n = 5), PCP (1.0mg/kg, n = 4) or the competitive NMDA antagonist CGS-19755 (1.8mg/kg, n = 4) from vehicle. Up to doses that caused large reductions in response rates, ethanol produced only vehicle-appropriate responding in the pigeons trained to PCP and only low levels of drug-appropriate responding in pigeons trained to MK-801 and CGS-19755. The present results suggest there could be asymmetric generalization between the discriminative stimulus effects of i.g. ethanol and NMDA antagonists.     

Does MK-801 discrimination constitute an animal model of schizophrenia useful for detecting atypical antipsychotics?

Two groups of female Wistar rats were trained to discriminate two doses (0.075 and 0.0375 mg/kg) of the noncompetitive NMDA antagonist MK-801 (dizocilpine) in a food-rewarded operant FR30 drug discrimination task. The atypical neuroleptic clozapine (2-6 mg/kg) produced only minimal antagonism (max. 32%) of the MK-801 cue at either training dose, and the "antagonist" effects were not clearly dose related. Furthermore, in the 0.075 mg/kg trained animals clozapine at 3 mg/kg failed to shift the MK-801 dose-response curve to the right. The alpha1-adrenoceptor antagonist prazosin (1-8 mg/kg) was also tested for antagonism of the 0.0375 mg/kg MK-801 cue, and again, only partial antagonism was seen (maximum 36%). Recently, it was suggested [4] that as the discriminative stimulus produced by MK-801 (0.075 mg/kg) was fully antagonized by clozapine at 3 mg/kg, but not by the typical neuroleptic haloperidol, this assay may be a useful screen for detecting atypical neuroleptics. It would seem, however, that this is not necessarily the case, and that the MK-801 discriminative cue may not be psychotomimetic. However, as this was a food rewarded rather than an avoidance paradigm that was used in the prior study [4], it may be that the drug discrimination procedure itself is a critical factor, although this hypothesis requires empirical testing.     

The discriminative stimulus properties of self-administered ethanol are mediated by GABAA and NMDA receptors in rats

RATIONALE: The neurobiological systems that mediate the discriminative stimulus effects of self-administered drugs are largely unknown. The present study examined the discriminative stimulus effects of self-administered ethanol. METHODS: Rats were trained to discriminate ethanol (1 g/kg, IP) from saline on a two-lever drug discrimination task with sucrose (10% w/v) reinforcement. Test sessions were conducted with ethanol (0 or 10% v/v) added to the sucrose reinforcement to determine if self-administered ethanol would interact with the discriminative stimulus effects of investigator-administered ethanol, or with the ethanol-like discriminative stimulus effects of the GABAA-positive modulator pentobarbital or the non-competitive NMDA antagonist MK-801. RESULTS: During a saline test session, ethanol (10% v/v) was added to the sucrose reinforcement. Responding by all animals began accurately on the saline-appropriate lever and then switched to the ethanol-appropriate lever after rats self-administered a mean dose of 1.2 +/- 0.14 g/kg ethanol. During cumulative self-administration trials, responding initially occurred on the saline lever and then switched to the ethanol-appropriate lever after ethanol (0.68 +/- 0.13 g/kg) was self-administered. Investigator-administered MK-801 (0.01-1.0 mg/kg, cumulative IP) and pentobarbital (0.3-10.0 mg/kg, cumulative IP) dose-dependently substituted for ethanol. When ethanol (10% v/v) was added to the sucrose reinforcer, MK-801 and pentobarbital dose-response curves were shifted significantly to the left. CONCLUSIONS: Self-administered ethanol substituted for and potentiated the stimulus effects of investigator-administered ethanol, suggesting that the discriminative stimulus effects of self-administered ethanol are similar to those produced by investigator-administered ethanol. Self-administered ethanol enhanced the ethanol-like discriminative stimulus effects of MK-801 and pentobarbital, which suggests that the discriminative stimulus effects of self-administered ethanol are mediated by NMDA and GABAA receptors.     

Generalisation of ethanol with drug mixtures containing a positive modulator of the GABA(A) receptor and an NMDA antagonist

Ethanol is thought to produce its discriminative stimulus effect by actions on two or more neurotransmitter systems. To test this idea further, rats were trained to discriminate mixtures of two drugs from vehicle in two-lever procedures with food reinforcers presented on a tandem variable-interval fixed ratio schedule. After drug-appropriate responding with the training mixtures reached 85%, generalisation to ethanol was examined in extinction tests. Rats trained to discriminate a mixture of chlordiazepoxide (5.0 mg/kg, s. c.) plus dizocilpine (0.08 mg/kg, i.p.) yielded a mean of 76% drug-appropriate responding when tested with ethanol (3.0 g/kg, i.g. ). However, when rats were trained with an 8.0 mg/kg dose of pentobarbitone in a mixture with 0.08 mg/kg of dizocilpine, the same dose of ethanol produced only 33% drug-appropriate responding. After retraining with pentobarbitone (12 mg/kg) plus dizocilpine (0.04 mg/kg), ethanol (3.0 g/kg, i.g.) produced 75% drug-appropriate responding. Pentobarbitone and dizocilpine administered alone produced full, dose-related generalisation, but there was no generalisation to (+)-amphetamine (0.025-0.8 mg/kg, s.c.). Thus, ethanol substituted for mixtures in which the GABA(A)-modulatory component had equal or greater salience than the NMDA-antagonist component. Doses of ethanol that generalised with the drug mixtures always reduced overall rates of responding as compared with control rates. Nevertheless, these data provide further support for the hypothesis that ethanol produces a compound stimulus comprised of elements resembling the effects of positive modulators of GABA(A) receptors and those of NMDA antagonists.     

Four-choice drug discrimination in pigeons

(+)Amphetamine was added as a training stimulus for pigeons previously trained to discriminate among pentobarbital, morphine and saline using a three-choice procedure. Pigeons quickly learned the four-choice drug discrimination. Generalization from the training drugs was similar to that established with simpler drug discriminations; pentobarbital generalized to chlordiazepoxide, morphine generalized to methadone, and (+)amphetamine generalized to cocaine and methamphetamine. Low doses of phencyclidine generalized to saline, while higher doses partially generalized to pentobarbital and (+)amphetamine. When dose-response curves were redetermined with a cumulative-dosing procedure, the same pattern of generalization occurred as under single-dose procedures. Dose-response curves were quantal under both the single-dose and the cumulative-dosing procedures. The four-choice procedure offers some important advantages for studying the discriminative stimulus effects of drugs that interact with multiple receptor subtypes and for studying drug mixtures.     

A role for receptors of N-methyl-D-aspartic acid in the discriminative stimulus properties of phencyclidine

Ketamine and (+)-N-allylnormetazocine ((+)-NANM) were found to generalize in a rat operant drug discrimination paradigm to the interoceptive stimulus induced by phencyclidine (PCP). Intraperitoneal administration of the non-competitive antagonist of the N-methyl-D-aspartate (NMDA) receptor, MK-801, and intracerebroventricular injection of the competitive antagonist, 2-DL-amino-7-phosphonoheptanoic acid (2-APH), also resulted in a dose-dependent generalisation to the PCP discriminative stimulus. The results suggest that NMDA receptor antagonism may play an important role in the mediation of the discriminative stimulus properties of PCP. The low potency of MK-801 and 2-APH to displace [3H](+)-NANM binding in vitro argues against an involvement of the haloperidol-sensitive sigma recognition site in the behaviour.     

The discriminative stimulus effects of MK-801: Generalization to other N-methyl-D-aspartate receptor antagonists

Rats were trained to discriminate a dose of the NMDA antagonist MK-801 from saline using a standard, two-lever, operant procedure. The acquisition of this discrimination was rapid, rats reaching criterion performance in a mean of 32 sessions. The discriminative stimulus produced by MK-801 generalized to phencyclidine and (+)N-allylnormetazocine but only low levels of generalization were seen with (-)N-allylnormetazocine and with the anti- ischaemic drug ifenprodil and its derivative SL 82.0715. The MK-801 stimulus was partially antagonized by haloperidol. The results show that MK-801 can produce a discriminative stimulus which is shared by other non-competitive NMDA antagonists which have similar behavioural effects in other procedures. However, ifenprodil and SL 82.0715, which have also been shown to antagonize the effects of NMDA, do not produce similar behavioural effects or discriminative stimuli presumably because they act through a different site.     

Antagonism of AND and AND-OR drug mixture discriminations in rats

It has been suggested that use of the AND-OR training method may be associated with an enhancement of the pharmacological specificity of discriminations based on mixture of drugs. Rats were trained to discriminate a mixture of nicotine (0.4 mg/kg s.c.) plus midazolam (0.2 mg/kg s.c.) from saline (AND-discrimination, n = 8) or to discriminate the mixture from either drug alone (AND-OR discrimination, n = 6). The studies used two-lever operant procedures with food reinforcers presented on a tandem schedule. After discriminations were acquired to 80% accuracy, the nicotine antagonist mecamylamine (0.03–1.0 mg/kg s.c.) and the benzodiazepine antagonist flumazenil (0.32–10 mg/kg i.p.) were tested on the response to the mixture of nicotine plus midazolam. The antagonist effects of either mecamylamine or flumazenil given alone were more marked in rats trained under the AND-OR procedure than in rats trained on the AND-discrimination. Similarly, the antagonist effects of mixtures of mecamylamine plus flumazenil were much more potent under the AND-OR than under the AND-discrimination procedure. The AND-OR method reduced the dose of the antagonist mixture needed to produce complete block by a factor of about 10, as compared with the AND-discrimination. These striking differences in sensitivity to antagonists support the view that AND-OR or related procedures may enhance the pharmacological specificity of complex drug discriminations.     

Strategies for understanding the pharmacological effects of ethanol with drug discrimination procedures.

Ethanol appears to produce a stimulus complex, or compound cue, composed of distinct components that are mediated by different receptor systems. In ethanol vs. water discriminations, it appears that ethanol produces a redundant stimulus complex such that separate, receptor-mediated activity can serve as the basis for the discrimination. These discriminations have been termed redundant, because multiple features of the cue could serve as the basis of the discrimination. In ethanol vs. water discriminations, one common feature is the asymmetrical generalizations between components of the ethanol cue and ethanol. There is also evidence for overshadowing of one component by other components of the ethanol stimulus complex. It appears possible to transfer the basis of the ethanol cue from a redundant cue to a conditional cue with specific training procedures. When the discriminative stimulus effects of ethanol are juxtoposed with those of one component of the ethanol complex, as in ethanol vs. water vs. pentobarbital discriminations, the ethanol discrimination shifts to a conditional basis. The ability to antagonize an ethanol discrimination may be dependent upon whether the discrimination is based on redundant component stimuli or conditional presence of all component stimuli.     

Amnesic effect of the novel anticonvulsant MK-801

MK-801, a reported N-methyl-D-aspartate (NMDA) antagonist with affinity for the phencyclidine (PCP) receptor, injected intravenously in mice before a training trial in a passive avoidance procedure, produced a similar amnesic effect to that produced by the standard amnesic agent scopolamine. Compared to vehicle-treated mice, each drug produced significant amnesia, yet the potency of MK-801 was 40 times that of scopolamine. This result with the MK-801 is consistent with previous reports that drugs which act at PCP recognition sites within the brain produce memory impairing effects in rodents.     

Discriminative stimulus effects of ethanol in rats using a three-choice ethanol-midazolam-water discrimination

Three-choice discrimination procedures are used to characterize how similar the discriminative stimulus effects of two drugs are in relation to each other. This procedure has suggested similarities between ethanol and ligands that positively modulate the gamma-aminobutyric acid type A (GABAA) receptor complex. As an extension to these studies, male Long-Evans rats were trained to discriminate midazolam (3.0 mg/kg, i.p.) from ethanol (1.0 g/kg, i.g.) from water (2.3 ml, i.g.) in a three-lever, food reinforced task. Substitution tests were conducted following administration of GABAA-positive modulators, noncompetitive N-methyl-D-aspartate (NMDA) antagonists, 5-HT1B agonists and isopropanol. Among the GABAA-positive modulators, diazepam was the only drug that completely substituted for midazolam; both pentobarbital and the neurosteroid allopregnanolone showed partial midazolam substitution. The NMDA antagonist dizocilpine substituted for ethanol, while phencyclidine showed no substitution for either ethanol or midazolam. The serotonin agonists tested also showed no substitution for either ethanol or midazolam. Isopropanol was the only other drug that completely substituted for ethanol. These data extend previous findings from an ethanol-pentobarbital-water discrimination and further define training conditions that result in a conditional basis for the ethanol discrimination where only those drugs with pharmacological heterogeneous effects similar to ethanol produce a full ethanol-like effect.     

Prefrontal dopamine is directly involved in the anxiogenic interoceptive cue of pentylenetetrazol but not in the interoceptive cue of chlordiazepoxide in the rat

Rationale: The prefrontal cortical (PFC) dopamine (DA) system has been implicated in anxiety-related behavioral changes, but direct, unequivocal support for this idea is sparse. Objectives: The present aim was to study the functional significance of prefrontal DA using the pentylenetetrazol (PTZ) discrimination model of anxiety. A comparison was made with its role in the cue of the anxiolytic drug chlordiazepoxide (CDP). Methods: Two groups of rats were trained to discriminate either PTZ (20 mg/kg, s.c.) or CDP (10 mg/kg, i.p.) from saline using an operant drug discrimination procedure. After prolonged training, half of each group was used to assess biochemical changes induced by both drugs in different sub areas of the PFC. For the remaining rats, discrimination training continued and generalization tests with PTZ and CDP were performed. Rats were then provided with bilateral guide cannulae aimed at the ventromedial (vm) PFC, and the effects of local infusions of DAergic drugs on discriminative performance were evaluated. Results: CDP did not affect PFC DA activity, but PTZ increased the DOPAC/DA ratio in the vmPFC selectively. Generalization tests showed that the cues of PTZ and CDP were dose dependent. In PTZ-trained rats, infusions of the DA receptor antagonist cis-flupenthixol into the vmPFC blocked the PTZ cue dose dependently, whereas the agonist apomorphine partially generalized to this cue. In CDP-trained rats, neither drug antagonized or generalized to the CDP cue, showing that PFC DA is not critically involved in the CDP cue and that local pharmacological manipulations of PFC DA do not affect discriminative abilities per se. Conclusions: The DAergic innervation of the PFC is directly involved in the behavioral effects of PTZ, suggesting a role for it in anxiety. Received: 26 July 1999 / Final version: 9 January 2000