Individual variability of dopamine release from nucleus accumbens induced by nicotine.

Effects of subcutaneous administration of vehicle, amphetamine (1 mg/kg) or nicotine (0.4 mg/kg, injected twice, 90 min apart) on extracellular dopamine (DA) concentration in the nucleus accumbens (ACC) and ventral tegmental area (VTA) of the Sprague-Dawley rat were studied using microdialysis. Experiments were conducted at least 10 days following implantation of guide cannulae, and at least 2 h following insertion of microdialysis probes into the guides on the morning of each experiment. Probes were perfused at 2.5 microl/min and several fractions were collected every 10 min before and after the two test injections. Samples were analyzed by high-performance liquid chromatography with electrochemical detection for the major neurotransmitters and their metabolites. Significant DA release following nicotine administration was observed in ACC but not in VTA. By classifying ACC DA responses of individual rats, three major subgroups were identified which exhibited more robust responses. Nicotine appeared to be acting as a modulator of ACC DA, increasing DA output if baseline was <5 nM, but slowing release when the baseline exceeded 5 nM. These data are consistent with previous reports of modulation of arousal level by nicotine via DA.

Creation and first 20 years of the society for the stimulus properties of drugs (SSPD).

Clinical observations and novels in the 19th century recognized that memory of some events can be retrieved only under the influence of the same drug condition that was present during the event. This dissociative effect of drugs probably reflects the same drug effects that were later called the discriminative stimulus effects of drugs. The Society for Stimulus Properties of Drugs (SSPD) was founded in 1978 as a forum for communications and periodic meetings on this drug effect. During its early years many of its members were psychologists, but subsequent to that time the most frequent research application has been for the pharmacological purpose of identifying new drugs that have the same discriminative stimulus attributes as a prototype training drug. The majority of members have been in the United States, but several major international meetings have been in Europe. The methods used by the society's members involve both neuropharmacological and psychological processes, allowing them to make unique contributions to the study of both mind and brain.

Discriminative stimulus (DS) properties of nicotine in the C57BL/6 mouse.

Previous research conducted in this and other laboratories has examined the role of genetic factors in determining sensitivity to (-)-nicotine in a variety of behavioral and physiological measures in the rat. More recent research further indicates that genetic factors can also influence the level of sensitivity to (-)-nicotine when serving as a discriminative stimulus (DS) in different rat strains. However, there has been little work examining the influence of genotype on the discriminative stimulus (DS) properties of (-)-nicotine in mice, a species that has played a major role in understanding the relationship between genetics and (-)-nicotine pharmacological effects. To further our understanding of the role of genetics and the ability of (-)-nicotine to exert DS control of behavior in the mouse, a group of C57BL/6 mice was trained to discriminate 0.4 mg/kg (-)-nicotine from saline using a two-lever operant procedure. (-)-Nicotine's discriminative stimulus in C57BL/6 mice appears to be similar to that generated in the rat. Results from behavioral tests with other drugs indicated that d-amphetamine exhibited a partial generalization, while (+)-nicotine fully generalized with nicotine. Tests of antagonism with mecamylamine and scopolamine further showed the cholinergic specificity of the (-)-nicotine DS in the mouse; mecamylamine but not scopolamine completely antagonized the (-)-nicotine DS. This work lays the groundwork for future comparisons of different mouse strain's sensitivities to (-)-nicotine's discriminative stimulus as well as using this behavioral model to search for new nicotinic receptor agonists and antagonists.

Effects of nicotine on the hypothalamic-pituitary-axis (HPA) and immune function: introduction to the Sixth Nicotine Round Table Satellite, American Society of Addiction Medicine Nicotine Dependence Meeting, November 15, 1997.

This meeting was the sixth consecutive Nicotine Round Table Satellite Meeting which was held in Washington, DC, on 15 November, 1996; previous meetings are presented in Table I. The overall objective of these meetings was to bring together scientists and clinicians as a means of developing a dialogue concerning the basic mechanisms of nicotine action and the effects of nicotine on the whole organism. The specific topic of this meeting was chosen because of the potent effects of nicotine on the hypothalamic-pituitary-axis (HPA), and newer concepts indicating that the immune system and the HPA are connected via a variety of neuroendocrine and neurotransmitter elements. Whereas the HPA appears to play a unique role in adapting to internal and external stressors, the immune system appears to act as a forward scout which provides information important to the HPA. This Satellite Meeting evaluated the effects of nicotine from three points of view: (1) the effects of nicotine on HPA function; (2) the effects of the HPA on the pharmacological effects of nicotine; and (3) the effects of nicotine on immune function. This specific presentation will provide an overview of the findings of the meeting and will discuss several of the overriding issues in this area of nicotine research.

Pharmacological investigation of (+)- and (-)-cis-2,3,3a,4,5,9b-hexahydro-1-methyl-1H-pyrrolo-[3,2-h]isoq uinoline, a bridged-nicotine analog.

We recently synthesized a bridged-nicotine (BN) analog and its enantiomers. They failed to compete for [3H]nicotine binding in rat brain homogenates, yet they produced nicotine-like effects by decreasing locomotor activity and producing antinociception in the tail-flick, hot-plate and PPQ tests in mice. Therefore, additional in vivo and in vitro studies were undertaken to determine whether these compounds are indeed acting independently of the nicotinic system. Although these analogs did not produce nicotine-like responding when evaluated in rat drug discrimination, the racemate augmented the cue when administered in conjunction with nicotine. Moreover, the antinociceptive measured in the different tests and hypothermic effects of (+)-BN, the more potent enantiomer, were not blocked by the nicotinic antagonists mecamylamine and dihydro-beta-erythroidine. Acute tolerance developed to (+)-BN-induced antinociception but not to hypothermia after subcutaneous administration. In addition, no cross-tolerance was observed between (+)-BN and nicotine in the different tests. The absence of generalization in the discrimination test suggests that the BN analogs do not possess nicotine-like activity. In addition, the failure of mecamylamine and dihydro-beta-erythroidine to antagonize the antinociceptive and hypothermic effects of (+)-BN, on one hand, and the inability of the bridge analogs to stimulate 86Rb+ efflux in brain synaptosomes, on the other hand, provide further evidence that BN analog agonist effects are not mediated by the alpha-4, beta-2 receptor subunit combination. It is unlikely that alpha-7 subunits mediate the agonists effects of BN analogs because their affinity to neuronal [125I]alpha-bungarotoxin binding sites is in the higher micromolar range. Other nicotinic receptor subtypes remain possible candidates because (+/-)-BN augments the generalization of nicotine in drug discrimination and produces some nicotine-like pharmacological effects. BN analogs could represent a novel class of nicotinic analgesics because naloxone and atropine failed to alter the antinociceptive effects of (+)-BN. Alternatively, their actions may be entirely independent of the nicotinic system.

Disruption of Th1/Th2 cytokine balance by cocaine is mediated by corticosterone.

Cocaine has been shown to affect immune function through the release of corticosterone. Acute administration of both cocaine and corticosterone produces an enhancement of the T-dependent antibody response to sheep erythrocytes. The T-independent antibody response to DNP-ficoll is not enhanced under identical conditions, suggesting that the T-cell is involved as a cellular target. We examined T-helper cell cytokine production following in vivo cocaine administration and found an increase in IL-4 and IL-10; while IL-2 and IFN-gamma were unaffected. The rise in Th2 cytokines is consistent with an enhanced T-dependent antibody response, a measure of humoral immunity. Because previous results showed that the enhancement by cocaine is mediated via corticosterone, the direct effects of corticosterone on Th1/Th2 in vitro cytokine production were investigated. Th1 cytokines, IL-2 and IFN-gamma, were dose-dependently suppressed by corticosterone at physiologic concentrations. In contrast Th2 cytokines, IL-4 and IL-10, exhibited a biphasic dose response curve, whereby an enhancement was observed at low doses, followed by suppression at higher doses. In order to determine the consequences of this apparent shift towards a Th2 response on a Th1 response, we looked at the delayed-type hypersensitivity response to sheep erythrocytes. This measure of cell-mediated immunity was not significantly affected by acute cocaine, however, corticosterone administration resulted in a significant suppression. These results indicate that corticosterone can produce a shift towards a Th2 predominate response, possibly at the expense of Th1-mediated responses.

Role of corticosterone in the enhancement of the antibody response after acute cocaine administration.

A model has been developed in which acute cocaine administration results in an enhanced T-dependent antibody response to sheep erythrocytes. This enhancement occurs when cocaine (30 mg/kg, twice in 1 day) is administered 1 or 2 days before sensitization with antigen, in mice older than 16 wk. Acute cocaine has been shown to elicit a rise in serum corticosterone, and the administration of exogenous corticosterone, under similar conditions as cocaine, also results in a similar immunoenhancement. Further evidence in support of a role by corticosterone is the lack of an enhancement in adrenalectomized mice and the ability of alpha-helical corticotropin releasing factor to block the enhancement by cocaine. The role of concomitant epinephrine release from the adrenal was addressed by adrenal demedullation. Eliminating epinephrine, but not corticosterone release, had no effect on the cocaine-induced immunoenhancement. The evidence presented provides support for a major role by corticosterone in mediating cocaine's effects on at least one measure of immune function, the T-dependent antibody response.

Comparative pharmacology of nicotine and ABT-418, a new nicotinic agonist.

ABT-418, a novel cholinergic ligand, was reported to possess potent cognitive-enhancing and anxiolytic properties in animal models with reduced side effects (Decker et al. 1994; Garvey et al. 1994) suggesting selectivity of effects. In this study, the binding properties of ABT-418 to [3H]-nicotine sites were evaluated and its pharmacology investigated in different tests in laboratory animals. ABT-418 binds with high affinity to 3H-nicotine binding sites in the brain with, however, a Ki (6 nM) less than that of nicotine (four-fold). In addition, it acts as a full nicotinic agonist in producing hypomotility, hypothermia and antinociception in mice and engendering nicotine-like responding in rat drug discrimination. The potency of ABT-418 is three to four times less than that of nicotine in all of the animal models, except for hypothermia. In addition, its behavioral effects are completely blocked by mecamylamine, a non-competitive nicotinic antagonist. Although activation of nicotinic receptors by ABT-418 produced several behavioral and pharmacological effects, our results do not suggest high selectivity of different effects as reported by Decker et al. (1994) and Garvey et al. (1994). However, it should be noted that we did not perform some of these tests that produced effects at low doses (Decker et al. 1994) and additional pharmacological studies are needed to establish its selectivity at multiple nicotinic receptors.

The psychopharmacological basis of nicotine's differential effects on behavior: individual subject variability in the rat.

Nicotine, the presumed active pharmacological agent in tobacco, produces variable effects on behavior that are at best described as "paradoxical" in nature. Thus, nicotine, via tobacco use in humans or nicotine administration in experimental animals, tends to transpose behavior depending on predrug baseline rates of behavior. High rates of behavior appear to be reduced, while low rates of behavior appear to be increased by nicotine. This work further proposes that nicotine's variable effects on behavior may be related to its capacity to act as a behavioral agonist and/or antagonist via its ability either to activate or to desensitize distinct central nicotinic acetylcholinergic receptors (nAChR's). Nicotine is portrayed as a neuronal modulating agent that can affect behavior contingent upon the genetic makeup of the individual subject being studied. Depending on the structure, function, and location of distinct nAChR's, nicotine appears to be able to induce a wide range of behavioral effects important to the tobacco user. However, this does not rule out the role the importance that other biogenic amine systems (i.e., serotonin or dopamine) may have in the genetics of tobacco use or nicotine's variable effects on behavior.

Nicotine-induced acute tolerance: studies involving schedule-controlled behavior.

The major goal of the present study was to examine acute tolerance to nicotine-induced disruption of operant behavior following a single, noncontingent injection. Rats were trained to lever press for food reinforcement under a fixed ratio-30 schedule. Once trained, rats were injected with either saline or nicotine (0.8 mg/kg) in their home cages. After either a 90- or 180-min delay, each rat was injected with nicotine (0.4 mg/kg) and placed in the operant chamber for a 30-min behavioral evaluation session. This experiment was replicated with slight modifications 1 week later. The results of the present study suggest that 0.8 mg/kg of nicotine produces acute tolerance to the response rate decreasing effects of 0.4 mg/kg of nicotine. Because the tolerance-producing dose of nicotine was injected while rats were not in the test environment, they did not have an opportunity to practice the target behavior while under the influence of the drug. Hence, the acute tolerance observed in this study appears to be, at least partly, pharmacological (vs. behavioral) in nature, and may be related to a desensitization of central nicotinic acetylcholinergic receptors (nAChRs).

Pharmacological effects of epibatidine optical enantiomers.

The pharmacology of synthetic D- and L-epibatidine, an alkaloid originally characterized from frog skin, were studied in different behavioral tests in mice and rats. The two enantiomers have potent antinociceptive activity in mice using the tail-flick test, with an ED50 of 6.1 and 6.6 micrograms/kg for L- and D-epibatidine respectively. Epibatidine enantiomers were 200 x more potent than L-nicotine as an antinociceptive agent in mice after s.c. administration. Their analgesic effect was blocked by mecamylamine but not naloxone, an opiate antagonist. Both D- and L-epibatidine have high affinity (Ki 54.7 and 55.0 pM, respectively) for [3H]nicotine binding site in rat brain. In addition, they reduced mice locomotor activity and body temperature in a dose-dependent manner. In rats trained with nicotine (0.4 mg/kg), epibatidine enantiomers engendered nicotine-like responding in a dose-related manner with an ED50 of 1.00 and 0.93 micrograms/kg for D and L, respectively. The discriminative effect of L- and D-epibatidine in rats was blocked by mecamylamine but not by hexamethonium. As in binding results, there was no significant enantioselectivity for these effects in our study.

Evidence that nicotine can acutely desensitize central nicotinic acetylcholinergic receptors.

Current concepts concerning nicotine's CNS mechanism(s) of action suggest that this drug produces its effects via an interaction at nicotinic-cholinergic receptors (nAChRs) sensitive to acetylcholine. In vitro research further suggests that, following its initial agonist effect, this cholinergic drug may also induce a rapid desensitization of the nAChR similar to that of acetylcholine, resulting in termination of its pharmacological effect. Research described in this paper provides evidence of this secondary desensitization process in vivo by demonstrating nicotine's ability to induce acute tolerance in a Discriminative Stimulus (DS) paradigm. The ability of nicotine (400 micrograms/kg, SC) to elicit DS control of behavior in a two-lever operant procedure was significantly reduced via a challenge dose (800 micrograms/kg, SC) of nicotine administered 15-180 min before the training dose. Twenty-three of 52 rats demonstrated this phenomenon. The time to develop acute tolerance varied, providing additional evidence that these effects may be contingent upon individual rat variability. In addition, physostigmine was also observed to induce a similar desensitization in a random population of desensitizing rats. Lastly, there were no differences between desensitizers and non-desensitizers in relation to the ability of mecamylamine (1000 micrograms/kg, SC) to antagonize the DS, while in both populations of rats scopolamine (100 micrograms/kg, SC) failed to antagonize the DS.

Neurobehavioral mechanisms of nicotine action: role in the initiation and maintenance of tobacco dependence.

Basic neuroscience research conducted over the last quarter century has provided us with much information concerning potential biobehavioral and neuromolecular mechanisms involved in the initiation and maintenance of tobacco dependence. Nicotinic-acetylcholinergic receptors (AChRs), in addition to having a primary locus on cholinergic neurons, appear to be also located on a variety of noncholinergic neurons (presynaptic and/or postsynaptic sites). Nicotine therefore appears to be able to affect a variety of neuronal pathways involved in behavioral reward and arousal processes, which appear paramount to tobacco dependence. Nicotine appears to have several unique properties at the cellular level that allow it to act both as an agonist and as a potential antagonist at select AChRs. Nicotine's ability to act as an agonist appears to be contingent on an action at nAChRs, which initially open a receptor-linked cation channel, eliciting the entrance of CA++ (or other cations) into the cell. Cation entrance into the cell, therefore, may be the cellular transducer of nicotine's behavioral and dependence-producing effects. Subsequent to this initial agonist effect, the nicotinic receptor is believed to undergo a refractory period, via a desensitization process, during which Ca++ is prevented from further entrance into the cell. It is this ability to induce receptor desensitization which seems central to nicotine's ability to act as an antagonist. The duration of nAChR desensitization may also be useful in explaining individual variability to nicotine's behavioral effects and may be related to the induction of acute and/or chronic tolerance in both animals and man. Nicotine-induced desensitization may also be important to relapse in the smoker if conditioned stimuli are able to provoke such mechanisms, which could lead to the need to smoke. Finally, a model is presented to account for individual smoking patterns and level of tobacco dependence which is partially based on the proposed cellular mechanisms of nicotine action and desensitization at the nAChR.

A characterization of nicotine-induced tolerance: evidence of pharmacological tolerance in the rat.

The present studies were conducted in order to confirm and extend previous findings that the mechanisms of tolerance to the behaviorally disruptive effects of nicotine in a operant model were primarily pharmacological. Both the traditional methodology employing the determination of dose response curves before and after chronic drug administration and a methodology which omits the generation of dose response curves were utilized in these investigations of nicotine-induced tolerance. Rats developed tolerance to both pre- and post-session administration of nicotine, suggesting that the mechanisms of tolerance to the disruptive effects of nicotine are primarily pharmacological. The mechanisms underlying these effects, however, remain to be evaluated.

Discriminative stimulus properties of nicotine: mechanisms of transduction.

Our thinking about how nicotine might be inducing DS control of behavior has changed drastically in the past 25 years. Our first inclination was that nicotine was mimicking ACh at a variety of specific and select n-AChRs. Then several nicotine researchers suggested that nicotine might be acting via specific and select noncholinergic receptors. At present, we seem to have returned to the view that nicotine may have pronounced effects at the n-AChR (figure 1), at least in some specific cases such as in the development of tolerance and in rats trained to discriminate nicotine (figure 6). The endogenous ligand has not changed, but the mechanism of how it affects presynaptic and postsynaptic receptors appears to have been rediscovered. The concept of rapid ACh-induced desensitization at the n-AChR is not new and appears basic to cholinergic neuronal function (figure 5). The desensitization concept has been revitalized by several investigators who also consider this mechanism important to how nicotine might act in protecting DA neurons from neurotoxicity of chemicals such as 6-OHDA or MPTP (Janson et al. 1988). The overall concept suggests that n-AChR desensitization at presynaptic DA sites may reduce neuronal accessibility to select exogenous neurotoxins and thus attenuate neuronal destruction. This hypothesis has also been partially validated in relation to the cholinergic neuron by providing preliminary evidence that nicotine was able to reduce cholinergic neuron destruction (measured by brain ACh levels) via the intraventricular administration of the neurotoxin AF64A, an ACh nitrogen mustard (Villanueva et al. 1990). Thus, nicotine or compounds acting like nicotine could possibly be beneficial to patients exhibiting the select neurological problems observed in Parkinson's and Alzheimer's diseases. The process of desensitization might also be useful to understanding why humans choose to smoke tobacco products. Perhaps the ability of nicotine to induce such neuronal effects at a specific n-AChR makes it reinforcing (or aversive) to behavior. Such a mechanism of action might explain why nicotine appears to both increase and decrease arousal levels in animals (or humans) exhibiting differential basal level of excitability (Hendry and Rosecrans 1982), or why some people never become dependent on nicotine. How nicotine alters the n-AChR may also be beneficial to our understanding of the subtle nature of cholinergic neuronal function in learning, memory, and other behavioral states. The ability of ACh (or nicotine) to induce an activation or attenuation of some cholinergic or noncholinergic neuron or both may be important to these brain processes.(ABSTRACT TRUNCATED AT 400 WORDS)

Identification of metabolites of 3,4-methylenedioxymethamphetamine in rats.

Liquid chromatography with electrochemical detection (LC/ECD) and gas chromatography/mass spectrometry (GC/MS) were used to identify metabolites of N-methyl-3,4-methylenedioxyamphetamine (MDMA) in samples of rat plasma and urine. Several potential metabolites, based on what is known about the metabolism of the desmethyl analog (i.e., MDA), were synthesized as standards to aid in the identification of the MDMA metabolites. MDA and N-methyl-1-(4-hydroxy-3-methoxy-phenyl)-2-aminopropane (3b) were identified in urine by HPLC and confirmed by GC/MS. 1-(4-Hydroxy-3-methyoxyphenyl)2-aminopropane, (3a), N-methyl-1-(3-hydroxy-4-methoxyphenyl)-2-aminopropane (2b) and 1-(3,4-dihydroxyphenyl)-2-aminopropane (4a) were tentatively identified by LC/ECD but insufficient sample size precluded confirmation by mass spectrometry. MDA was also identified in brain and plasma extracts. Because MDA is a metabolite of MDMA in humans, and because it has been speculated that the neurotoxic effects of MDA and MDMA may be due to a metabolite, the results of the present study may ultimately aid our understanding of the neurotoxic mechanism of these drugs of abuse.

Withdrawal from chronic nicotine fails to produce a conditioned taste aversion to saccharin in rats.

Sprague-Dawley rats were maintained on a daily regimen of nicotine, morphine or saline administration for 28 days. Following the discontinuation of the daily drug regimen, rats were given a choice of tap water or a saccharin-water solution. The rats previously receiving morphine drank significantly less saccharin-water solution than did the rats receiving nicotine or saline injections. The failure of the nicotine rats to display a conditioned aversion to the novel saccharin flavor suggests that nicotine did not produce a physiological withdrawal syndrome analogous to morphine withdrawal in this paradigm.