Serotonin involvement in the discriminative stimulus effects of kappa opioids in pigeons.

The purpose of the present study was to examine the role of serotonin (5HT) in the discriminative stimulus effects of kappa opioids. Pigeons were trained to discriminate 5.6 mg/kg of the kappa opioid, U50,488, from water. During substitution tests, both U50,488 and another kappa opioid, spiradoline, produced > 80% responding on the U50,488-appropriate key. In contrast, the non-opioid compound, phencyclidine and several serotonergic compounds failed to substitute for the U50,488 discriminative stimulus across a wide range of doses. During combination tests, the selective 5HT1A agonist, 8-OH-DPAT (0.001-3.2 mg/kg), dose-dependently attenuated the discriminative stimulus effects of 5.6 mg/kg U50,488 and 3.2 mg/kg spiradoline. This effect was reversed by the 5HT1A antagonist, NAN-190 (0.01-1 mg/kg), in a dose-dependent manner. Buspirone (0.01-10 mg/kg), a 5HT1A partial agonist, also attenuated the discriminative stimulus effects of the training dose of U50,488 but ipsapirone, another 5HT1A partial agonist, did not. Ketanserin, a 5HT2 antagonist, and MDL72222, a 5HT3 antagonist, attenuated the effects of U50,488, whereas the 5HT1B,1C agonist, mCPP, and the 5HT2 agonist, DOI, did not. Depletion of 5HT with p-CPA also attenuated U50,488's discriminative stimulus effects. Taken together, the results suggest that serotonin release is an important component in the discriminative stimulus effects produced by kappa opioids; however, the effects of DOI and mCPP alone suggest that activation of post-synaptic 5HT receptors is not sufficient to produce the full spectrum of kappa opioid discriminative stimulus effects.

Effect of once weekly treatment with 3,4-methylenedioxymethamphetamine on schedule-controlled behavior in rats.

The present study examined the effects of 3,4-methylenedioxymethamphetamine (MDMA), before and after once a week dosing, on the behavior of rats responding under a fixed ratio 20 schedule of reinforcement. Acutely, cumulative doses of MDMA dose-dependently decreased responding when compared to a series of water injections. Rats were then separated into two groups, one of which received only weekly MDMA ('paired') while the other received an additional injection of water each week ('unpaired'). Weekly dosing with MDMA resulted in significantly increased responding at low doses in the paired group but not in the unpaired group. When water injections were readministered there was a significant increase in responding in both groups. During the weekly regimen, locomotor activity also increased significantly over time after both water and MDMA injections. In conclusion, it appears that even weekly dosing with a small amount of MDMA can have long-lasting effects that are manifested in both operant and spontaneous behavior and that may be mediated by a conditioning mechanism.

Behavioral and developmental effects of two 3,4-methylenedioxymethamphetamine (MDMA) derivatives.

The effects of 3,4-methylenedioxymethamphetamine (MDMA or 'ecstacy') and two structurally related compounds, N-methyl-1-(3,4-methylenedioxyphenyl)-1-ethanamine (MDM1EA) and N-methyl-1-(3,4-methylenedioxyphenyl)-3-butanamine (HMDMA) were examined in two preparations: (i) a drug discrimination procedure in MDMA-trained rats and (ii) the chicken embryo, for determination of the direct effects of these compounds on the developing organism. The highest doses of MDM1EA and HMDMA partially substituted for MDMA, whereas higher (30-60 mg/kg) doses of HMDMA evoked clonic seizures in a separate group of rats. In chicken embryos MDMA had no effect on body, brain or liver weight, while the highest dose of MDM1EA decreased body weight and the 2 lowest doses of HMDMA increased body weight. All doses of HMDMA decreased liver weight (expressed as % body weight) when compared with contemporaneous water-treated controls. Taken together, the results of these experiments suggest that structurally related compounds share some stimulus properties with MDMA and may therefore share abuse liability. Furthermore, both MDMA-related compounds produced adverse effects on the developing organism, whereas MDMA did not.

Effects of designer drugs on the chicken embryo and 1-day-old chicken.

The present study was conducted to examine the effects of d-amphetamine and the designer drugs 3,4-methylenedioxymethamphetamine (MDMA), N-methyl-3,4-methylenedioxyphenyl-3-butamine (HMDMA), 3,4-methylenedioxyphenyl-2-butanamine (BDB), 3,4-methylenedioxyphenyl-1-ethanamine (MDM1EA) in the chick embryo and the young chicken. HMDMA and MDM1EA had no effect on motility on day 14 of embryogenesis, while MDMA, BDB, and d-amphetamine decreased embryonic motility at one or more doses. On day 1 posthatch, chickens were challenged with cumulative injections of water or the same drug that they had received in ova. With the exception of MDM1EA, all of the drugs produced effects such as distress vocalization, wing extension, tremor, flat body posture, bursting forward movements, loss of righting reflex, and convulsant-like kicking. Pretreatment with drug in ova resulted in tolerance to certain drug effects and supersensitivity to other drug effects. Furthermore, BDB significantly decreased hatchability, MDM1EA decreased body weight, and HMDMA decreased liver weight. Further studies are needed to determine the mechanism(s) of toxicity in this species.

Withdrawal from chronic haloperidol substitutes for the pentylenetetrazol discriminative stimulus.

The present study was designed to examine withdrawal from a therapeutic, non abused drug, haloperidol. Rats were trained to discriminate the anxiogenic compound pentylenetetrazol (PTZ) from water in a two lever, food reinforced, drug discrimination procedure. Dose effect curves were then determined for PTZ and the antipsychotic drug, haloperidol (0.1-1 mg/kg). Haloperidol did not substitute for PTZ, even at a dose that decreased rates of responding to approximately 15% of control values. Rats were then treated chronically with either 1 or 2 mg/kg/day haloperidol while training was suspended. After 5 days of chronic haloperidol 4/6 animals in the 1 mg/kg/day group and 5/7 in the 2 mg/kg/day group chose the PTZ lever when tested 24-48 hours after the last haloperidol injection. Haloperidol, 1 or 2 mg/kg, did not reverse PTZ-lever responding. After an additional 5 days of chronic haloperidol, 3/6 rats in the 1 mg/kg/day group and 5/7 rats in the 2 mg/kg/day group responded on the PTZ lever 24 hours after the last injection, and this was reversed with the anxiolytic, chlordiazepoxide (3.2-5.6 mg/kg). The current findings indicate that there is an anxiogenic component to withdrawal from haloperidol. In psychotic patients, abrupt discontinuation of haloperidol results in nausea, vomiting and sweating, as well as a "relapse into psychosis" characterized by anxiety, depression and internal chaos (1). Interestingly, the authors caution that the so-called relapse into psychosis may simply be a sign of withdrawal. The current findings support their view and suggest that abrupt discontinuation of psychoactive therapeutic agents may result in anxiety.

Withdrawal from chronic phencyclidine produces a pentylenetetrazol-like discriminative stimulus.

Rats trained to discriminate the anxiogenic compound pentylenetetrazol (PTZ) from water were implanted with osmotic mini-pumps containing 7 mg/kg/day phencyclidine (PCP) or water. Rats were tested for generalization to PTZ 24 hours prior to pump removal, and 4 to 96 hours after pump removal. While the pumps were in place, rats did not generalize to PTZ. When the pumps were removed on day 10, rats in the water group did not generalize to PTZ, but 69% of the rats in the chronic PCP group responded on the PTZ lever at 4 and/or 24 hours after pump removal, suggesting that the PCP withdrawal state mimics the interoceptive cue produced by PTZ. This withdrawal phenomenon was repeatable, in that rats that generalized once to PTZ during PCP withdrawal, generalized a second time when the procedure was repeated. In addition, the phenomenon was dose-dependent, as rats that did not generalize to PTZ after 7 mg/kg/day PCP did generalize when the chronic dose of PCP was increased to 10 mg/kg/day. These findings suggest that there is an anxiogenic component of PCP withdrawal and that tolerance does not develop to this effect.

Chronic bretazenil produces tolerance to chlordiazepoxide, midazolam, and abecarnil.

The purpose of the present study was to determine if chronic treatment with a nonsedative benzodiazepine partial agonist would confer tolerance to the rate-decreasing effects of other benzodiazepine ligands in a fixed-interval procedure in rats. A separate group of rats was treated chronically with the sedative benzodiazepine full agonist, chlordiazepoxide, for comparison. It was hypothesized that tolerance would develop rapidly to chlordiazepoxide due to loss of reinforcement density at rate-decreasing doses and that there would probably be cross-tolerance to other rate-decreasing benzodiazepine ligands such as midazolam and abecarnil. Because bretazenil does not produce rate decreases, however, it was not expected that tolerance would be found to chlordiazepoxide, midazolam, or abecarnil. After 8-12 weeks of chronic treatment with either chlordiazepoxide or bretazenil, however (final dose of benzodiazepine = 30 mg/kg/day), tolerance was found to the rate-decreasing effects of chlordiazepoxide, midazolam, and abecarnil in both groups. It is concluded that such tolerance was most likely due to a saturation of benzodiazepine receptors by this high-affinity partial agonist.

Chlordiazepoxide, but not bretazenil, produces acute dependence, as evidenced by disruptions in schedule-controlled behavior.

The purpose of the present study was to determine whether the full benzodiazepine (BDZ) agonist chlordiazepoxide (CDAP) and the partial BDZ agonist bretazenil would produce acute dependence in rats, as evidenced by disruptions in fixed-interval responding during precipitated abstinence withdrawal. Doses of CDAP and bretazenil administered acutely were 10, 75, and 100 mg/kg; flumazenil (1-56 mg/kg) was administered 1, 2, 4, or 18 h later. Withdrawal, defined as a significant decrease in fixed-interval responding, was only seen when a high dose of flumazenil was administered 18 h after 100 mg/kg of CDAP. These results support those of others (5) who found that high (75-450 mg/kg) doses of CDAP were required to produce acute physical dependence. That bretazenil did not produce acute physical dependence supports the findings of others (20,23) who report that chronic administration of bretazenil does not result in physical dependence.

Effects of mu, kappa and sigma opioids on fixed consecutive number responding in rats.

Responding in rats was maintained under a fixed consecutive number 20 schedule. Under this schedule, at least 20 consecutive responses were required on one lever before a response on a second lever produced food. Morphine, buprenorphine, ethylketocyclazocine (ECK), N-allylnormetazocine (NANM) and d-cyclazocine all caused dose-dependent decreases in response rate. With the exception of buprenorphine and EKC, each drug also produced a decrease in the percent of reinforced runs. Differences among the drugs were more apparent, however, on the basis of the conditional probability of switching to the second lever after any given run length on the first lever. Morphine increased the probability of premature switching and decreased the probability of switching after run lengths greater than 20. Buprenorphine decreased the probability of switching at all run lengths and EKC produced occasional increases in premature switching. In sharp contrast to the other opioids tested, NANM and d-cyclazocine generally increased the probability of switching at all run lengths. Thus, it appears that the fixed consecutive number schedule may be a sensitive procedure for distinguishing among the behavioral effects of various opioid agonists.

Evidence of single dose opioid dependence in 12- to 14-day-old chicken embryos.

We have previously reported that chicken embryos injected with a single dose of methadone (Meth) on day 3, 7 or 11 of embryogenesis fail to show dependence on day 14, measured as a significant overshoot in motility above baseline after challenge with the opioid antagonist naloxone (Nx). Constant infusion of Meth from day 7 to 14 also failed to produce evidence of dependence on day 14. To address the question of whether the 14-day-old embryo is capable of expressing withdrawal, isobutylmethylxanthine (IBMX), a compound that produces quasi-opioid withdrawal, was injected directly into the embryo, resulting in a significant increase in motility. To determine whether the 14-day-old embryo could also express true opioid withdrawal, the embryos were injected with various doses of Meth or morphine (Morph), followed at different time intervals by injections of varying doses of Nx. A high dose of Morph followed 24 hours later by a low dose of Nx produced evidence of withdrawal, as did a low dose of Meth followed 1 hour later by a higher dose of Nx, U50488H, a selective kappa agonist, had no effect on motility in the 14-day-old embryo, suggesting that the decrease in motility seen after Meth was not mediated by a kappa receptor. Pretreatment with the irreversible mu antagonist, beta-funaltrexamine (B-FNA), blocked the decrease in motility seen after Meth and also prevented the overshoot in motility when Nx was given 1 hour post-Meth. We were also able to demonstrate dependence/withdrawal in the 12-day-old embryo, but higher doses of both Meth and Nx were required.(ABSTRACT TRUNCATED AT 250 WORDS)

A behavioral comparison of Nexus, cathinone, BDB, and MDA.

The effects of 4-bromo-2,5-dimethoxyphenethylamine (Nexus), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxyphenyl-2-butanamine (BDB), and cathinone were studied in the newly hatched chicken and compared to the effects of d-amphetamine and three hallucinogens in the same species. Cathinone, a psychomotor stimulant in man (6), produced effects that were qualitatively similar to effects seen after administration of d-amphetamine (i.e., distress vocalization, wing extension, inability to stand, and loss of righting reflex). BDB, a compound with unknown activity in man, and two known hallucinogens, Nexus (5) and MDA (1), produced effects in the chicken that are common to both stimulants and hallucinogens in this species. For example, both MDA and BDB produced abnormal body posture that was identical to that reported after administration of hallucinogens such as lysergic acid diethylamide (LSD) and harmine (11). Nexus, on the other hand, produced rigid penguin-like posture, an effect seen in the chicken after administration of another hallucinogen, mescaline (12). BDB also produced bursting forward movements, an effect commonly observed after LSD and harmine. Our findings suggest that the young chicken can be used as an alternative, nonmammalian, model for predicting classification of new compounds.

Structure-activity relationships of BDB and its monomethyl and dimethyl derivatives.

The purpose of the present study was to evaluate the behavioral effects of 3,4-methylenedioxyphenyl-2-butanamine (BDB), N-methyl BDB (MBDB), and N,N-dimethyl BDB (MMBDB) in the newly hatched chicken. The primary amine, BDB, produced effects that are commonly seen in the chicken after administration of both hallucinogens and psychomotor stimulants (i.e., distress vocalization, tremor, and wing extension). It also produced abnormal body posture and bursting forward locomotion, effects elicited only by hallucinogens. Loss of righting reflex also occurred at the highest (16 mg/kg) dose of BDB, and this effect is typical of d-amphetamine but has not been reported for hallucinogens. The monomethylated derivative of BDB, MBDB, was less potent than BDB, and the N,N-dimethyl analogue of BDB, MMBDB, had no effect on behavior at the doses tested.

Tolerance/cross-tolerance to the discriminative stimulus effects of chlordiazepoxide and bretazenil.

Rats were trained to discriminate 10 mg/kg chlordiazepoxide (CDAP) from saline in a two-lever drug discrimination procedure. Both CDAP and the nonsedative benzodiazepine partial agonist, bretazenil, dose-dependently substituted for the training dose of CDAP. Training was then suspended and half of the rats were placed on chronic CDAP, while the other half received water. Tolerance to the discriminative stimulus effects of CDAP developed after 1 mo and a final dose of approximately 110 mg/kg/d, as evidenced by the fact that the training dose of CDAP no longer produced drug-appropriate responding. An insurmountable tolerance also developed to bretazenil, as no rat responded on the drug-appropriate lever at doses as high as 56 mg/kg, whereas in the prechronic dose-effect curve, 1 mg/kg of bretazenil produced 100% drug-appropriate responding. One week after chronic CDAP was discontinued, the dose-effect curve for CDAP in the chronic CDAP group was comparable to that obtained in the prechronic phase, indicating that the rats were no longer tolerant to CDAP. In contrast to CDAP, the dose-effect curve for bretazenil did not return to its prechronic level, with higher doses being required for substitution. In the chronic water group, the dose-effect curves for CDAP and bretazenil were essentially the same before, during, and after the chronic regimen. Thus, suspension of training for 6 wk does not result in loss of the discriminative stimulus control. Chronic exposure to CDAP, however, does result in tolerance to the discriminative stimulus effects of CDAP, with an accompanying insurmountable cross-tolerance to the partial benzodiazepine agonist, bretazenil. These results support findings by other investigators who find tolerance to the discriminative stimulus properties of a wide range of psychoactive drugs (Young and Sannerud, 1989).

Ultrawideband radiation and pentylenetetrazol-induced convulsions in rats.

New non-ionizing pulsed systems using ultrawideband (UWB) require safety assessment before they can be used by either military or civilian communities. The development of directed energy weaponry intended for use against electronically vulnerable targets, as well as ground-probing radar systems, have used fast-rise-time high-peak-power electromagnetic pulses characteristic of UWB emitters. It has been postulated that these ultrashort pulses might produce electromagnetic transients resulting in tissue damage. Several challenges to this notion have been posed, however. One report found that rats exposed to UWB after receiving a convulsant drug tended toward longer latency to the onset of convulsions than the no-exposure group. Although not statistically significant, the presence of this trend prompted the present study. An ED99 dose of the convulsant pentylenetetrazol (PTZ) or saline was given just before UWB or sham exposure and resultant seizure activity was recorded. The data from the current study show no effect of UWB exposure on PTZ-induced seizure activity, thereby not supporting the tissue damage concerns, at least for the exposure parameters used here.