Exposure to nicotine enhances its subsequent self-administration: contribution of nicotine-associated contextual stimuli.

Contextual stimuli present during nicotine exposure can come to act as conditioned stimuli and have been shown to play an important role in ongoing nicotine self-administration. In the present study, we characterized the effects of contextual stimuli previously paired with non-contingent nicotine exposure injections on subsequent nicotine self-administration. Rats were exposed to five injections of either saline or nicotine (0.4 mg/kg, i.p.) in either their home cage or a self-administration chamber with the levers retracted. Two weeks later, they were allowed to self-administer nicotine (30 µg/kg/infusion, IV) under fixed ratio (FR) schedules of reinforcement across 12 consecutive sessions. Lastly, responding under a progressive ratio (PR) schedule was assessed. Rats exposed to nicotine in the self-administration chamber subsequently increased their intake of nicotine across the FR test days, obtaining more infusions on average by days 7-12 compared to their saline exposed controls. This increase was not due to nicotine exposure alone as rats exposed to nicotine in the home cage did not show this effect. It was also not due to differences in the final ratio achieved between nicotine and saline exposed rats. Although rats exposed to nicotine in the self-administration chambers displayed reduced discrimination between the active and inactive levers during FR testing, they showed increased motivation to self-administer nicotine under the PR schedule. These results indicate that exposure to nicotine can enhance its subsequent self-administration and highlight the contribution of nicotine-associated contextual stimuli to the work output rats ultimately emit to obtain the drug.

Persistent reversal of enhanced amphetamine intake by transient CaMKII inhibition.

Amphetamine exposure transiently increases Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) α expression in the nucleus accumbens (NAcc) shell and this persistently increases local GluA1 S831 phosphorylation and enhances behavioral responding to the drug. Here we assessed whether transiently interfering with CaMKII signaling using a dominant-negative CaMKIIα mutant delivered to the NAcc shell with herpes simplex viral vectors could reverse these long-lasting biochemical and behavioral effects observed following exposure to amphetamine. As expected, transient expression of CaMKIIα K42M in the NAcc shell produced a corresponding transient increase in CaMKIIα and decrease in pCaMKIIα (T286) protein levels in this site. Remarkably, this transient inhibition of CaMKII activity produced a long-lasting reversal of the increased GluA1 S831 phosphorylation levels in NAcc shell and persistently blocked the enhanced locomotor response to and self-administration of amphetamine normally observed in rats previously exposed to the drug. Together, these results indicate that even transient interference with CaMKII signaling may confer long-lasting benefits in drug-sensitized individuals and point to CaMKII and its downstream pathways as attractive therapeutic targets for the treatment of stimulant addiction.

Intermittent nicotine exposure upregulates nAChRs in VTA dopamine neurons and sensitises locomotor responding to the drug.

Dopaminergic projections from the ventral tegmental area (VTA) to the nucleus accumbens (NAcc) mediate the behavioral and motivational effects of many drugs of abuse, including nicotine. Repeated intermittent administration of these drugs, a pattern often associated with initial drug exposure, sensitises the reactivity of dopamine (DA) neurons in this pathway, enhances the locomotor behaviors the drugs emit, and promotes their pursuit and self-administration. Here we show that activation of nicotinic acetylcholine receptors (nAChRs) in the VTA, but not the NAcc, is essential for the induction of locomotor sensitisation by nicotine. Repeated intermittent nicotine exposure (4 × 0.4 mg/kg, base, i.p., administered over 7 days), a regimen leading to long-lasting locomotor sensitisation, also produced upregulation of nAChRs in the VTA, but not the NAcc, in the hours following the last exposure injection. Functional nAChR upregulation was observed selectively in DA but not GABA neurons in the VTA. These effects were followed by long-term potentiation of excitatory inputs to these cells and increased nicotine-evoked DA overflow in the NAcc. Withdrawal symptoms were not observed following this exposure regimen. Thus, intermittent activation and upregulation by nicotine of nAChRs in DA neurons in the VTA may contribute to the development of behavioral sensitisation and increased liability for nicotine addiction.

α-1 Adrenergic receptors are localized on presynaptic elements in the nucleus accumbens and regulate mesolimbic dopamine transmission.

Brainstem noradrenergic neurons innervate the mesocorticolimbic reward pathway both directly and indirectly, with norepinephrine facilitating dopamine (DA) neurotransmission via α1-adrenergic receptors (α1ARs). Although α1AR signaling in the prefrontal cortex (PFC) promotes mesolimbic transmission and drug-induced behaviors, the potential contribution of α1ARs in other parts of the pathway, such as the ventral tegmental area (VTA) and nucleus accumbens (NAc), has not been investigated before. We found that local blockade of α1ARs in the medial NAc shell, but not the VTA, attenuates cocaine- and morphine-induced locomotion. To determine the neuronal substrates that could mediate these effects, we analyzed the cellular, subcellular, and subsynaptic localization of α1ARs and characterized the chemical phenotypes of α1AR-containing elements within the mesocorticolimbic system using single and double immunocytochemical methods at the electron microscopic (EM) level. We found that α1ARs are found mainly extra-synaptically in axons and axon terminals in the NAc and are enriched in glutamatergic and dopaminergic elements. α1ARs are also abundant in glutamatergic terminals in the PFC, and in GABA-positive terminals in the VTA. In line with these observations, microdialysis experiments revealed that local blockade of α1ARs attenuated the increase in extracellular DA in the medial NAc shell following administration of cocaine. These data indicate that local α1ARs control DA transmission in the medial NAc shell and behavioral responses to drugs of abuse.

Previous exposure to nicotine enhances the incentive motivational effects of amphetamine via nicotine-associated contextual stimuli.

The effect of nicotine exposure on the subsequent self-administration of amphetamine, extinction of this behavior, and amphetamine-induced reinstatement of drug seeking was assessed with particular attention to the contribution of contextual stimuli paired or unpaired with nicotine during exposure. Rats were exposed to five injections, one injection every third day, of either saline or nicotine (0.4 mg/kg, IP, base) in three experiments. In one, exposure injections were administered in the home cage. In another, they were administered in the self-administration chambers with the levers retracted. In a third, nicotine was administered either explicitly paired or unpaired with the self-administration chambers using a discrimination learning procedure. Starting 13-15 days later, rats were trained to self-administer amphetamine (100 µg/kg/infusion, IV), tested under a progressive ratio (PR) schedule for 6 days, subjected to up to 20 days of extinction training, and were then tested for reinstatement by non-contingent injections of amphetamine (0, 0.2, 0.4, and 0.75 mg/kg, IP). Nicotine enhanced the self-administration of amphetamine under the PR schedule and amphetamine-induced reinstatement but only when rats were tested in the chamber in which they were previously exposed to nicotine. These effects were not observed in rats exposed to nicotine in the home cage or in rats exposed to nicotine explicitly unpaired with the self-administration chambers. Exposure to nicotine also rendered rats resistant to extinction when amphetamine was withheld but this effect was observed regardless of nicotine exposure context, suggesting a separate consequence of drug exposure. Together, these results show that previous exposure to nicotine can enhance the incentive motivational effects of other psychostimulants like amphetamine and indicate a critical role for nicotine-associated contextual stimuli in the mediation of this effect. These findings have important implications for the treatment of addictions in humans.

Previous exposure to delta9-tetrahydrocannibinol enhances locomotor responding to but not self-administration of amphetamine.

Previous exposure to amphetamine leads to enhanced locomotor and nucleus accumbens (NAcc) dopamine (DA) responding to the drug as well as enhanced amphetamine self-administration. Here, we investigated the effects of exposure to Δ(9)-tetrahydrocannibinol (Δ(9)-THC) on behavioral and biochemical responding to amphetamine. Rats in different groups received five exposure injections of vehicle or one of five doses of Δ(9)-THC (0.4, 0.75, 1.5, 3.0, and 6.0 mg/kg i.p.) and were tested 2 days and 2 weeks later. Exposure to all but the lowest and highest doses of Δ(9)-THC enhanced the locomotor response to amphetamine (0.75 mg/kg i.p.), but all failed to enhance NAcc DA overflow in response to the drug. Moreover, exposure to 3.0 mg/kg i.p. Δ(9)-THC increased forskolin-evoked adenylyl cyclase activity in the NAcc and rats' locomotor response to the direct DA receptor agonist apomorphine (1.0 mg/kg s.c.), suggesting that Δ(9)-THC sensitized locomotor responding to amphetamine by up-regulating postsynaptic DA receptor signaling in the NAcc. Finally, amphetamine self-administration (200 µg/kg/infusion i.v.) was enhanced in amphetamine (5 × 1.5 mg/kg i.p.)-exposed rats, but not in rats exposed to Δ(9)-THC (5 × 3.0 mg/kg i.p.). Previous exposure to this dose of Δ(9)-THC modestly increased apomorphine SA (0.5 mg/kg/infusion i.v.). Thus, unlike amphetamine exposure, exposure to Δ(9)-THC does not enhance the subsequent NAcc DA response to amphetamine or promote amphetamine self-administration. Although Δ(9)-THC leads to alterations in postsynaptic DA receptor signaling in the NAcc and these can affect the generation of locomotion, these neuroadaptations do not seem to be linked to the expression of enhanced amphetamine self-administration.

Ascorbate modulation of sensorimotor processing in striatum of freely moving rats.

The striatum, which receives projections from the entire cortical mantle, is highly responsive to sensorimotor activity. Because either systemic or intra-striatal injections of ascorbate (AA) influence behavior known to involve striatal circuits, it is possible that the level of striatal AA, which is known to fluctuate with behavioral activation, directly alters striatal neuronal processing. To test this hypothesis, we recorded the activity of 94 presumed medium spiny striatal neurons in behaving rats treated with AA or vehicle and examined firing rate during periods of quiescence and sensorimotor stimulation (e.g., stroking of the whiskers, mid-back, and rump). Slow-scan voltammetry was used in separate rats to determine the extent to which AA treatment elevated striatal AA. Vehicle-treated rats had relatively slow basal firing rates at rest that routinely increased during sensorimotor stimulation. Comparable results were obtained in rats treated with 100 mg/kg AA, which failed to alter AA levels in striatum. Dose-dependent increases in striatal AA, however, occurred after injection of 500 and 1000 mg/kg AA, and at these doses, there was a significant decrease in the number of sensorimotor-related excitations. In fact, treatment with 1000 mg/kg AA reversed a significant proportion of excitations to inhibitions. Our results substantiate the role of the striatum in sensorimotor processing and emphasize extracellular AA as a modulator of striatal neuronal function.