Giovanni Hernandez

Prolonged rewarding stimulation of the rat medial forebrain bundle: Neurochemical and behavioral consequences.

Extracellular dopamine levels were measured in the rat nucleus accumbens by means of in vivo microdialysis. Delivery of rewarding medial forebrain bundle stimulation at a low rate (5 trains/min) produced a sustained elevation of dopamine levels, regardless of whether train onset was predictable. When the rate of train delivery was increased to 40 trains/min, dopamine levels rose rapidly during the first 40 min but then declined toward the baseline range. The rewarding impact of the stimulation was reduced following prior delivery of stimulation at the high, but not the low, rate. These results support the idea that dopamine tone plays an enabling role in brain stimulation reward and is elevated similarly by predictable and unpredictable stimulation.

At What Stage of Neural Processing Does Cocaine Act to Boost Pursuit of Rewards

Dopamine-containing neurons have been implicated in reward and decision making. One element of the supporting evidence is that cocaine, like other drugs that increase dopaminergic neurotransmission, powerfully potentiates reward seeking. We analyze this phenomenon from a novel perspective, introducing a new conceptual framework and new methodology for determining the stage(s) of neural processing at which drugs, lesions and physiological manipulations act to influence reward-seeking behavior. Cocaine strongly boosts the proclivity of rats to work for rewarding electrical brain stimulation. We show that the conventional conceptual framework and methods do not distinguish between three conflicting accounts of how the drug produces this effect: increased sensitivity of brain reward circuitry, increased gain, or decreased subjective reward costs. Sensitivity determines the stimulation strength required to produce a reward of a given intensity (a measure analogous to the KM of an enzyme) whereas gain determines the maximum intensity attainable (a measure analogous to the vmax of an enzyme-catalyzed reaction). To distinguish sensitivity changes from the other determinants, we measured and modeled reward seeking as a function of both stimulation strength and opportunity cost. The principal effect of cocaine was a two-fourfold increase in willingness to pay for the electrical reward, an effect consistent with increased gain or decreased subjective cost. This finding challenges the long-standing view that cocaine increases the sensitivity of brain reward circuitry. We discuss the implications of the results and the analytic approach for theories of how dopaminergic neurons and other diffuse modulatory brain systems contribute to reward pursuit, and we explore the implications of the conceptual framework for the study of natural rewards, drug reward, and mood.

Endocannabinoids Promote Cocaine-Induced Impulsivity and Its Rapid Dopaminergic Correlates

BACKGROUND Impaired decision making, a hallmark of addiction, is hypothesized to arise from maladaptive plasticity in the mesolimbic dopamine pathway. The endocannabinoid system modulates dopamine activity through activation of cannabinoid type 1 receptors (CB1Rs). Here, we investigated whether impulsive behavior observed following cocaine exposure requires CB1R activation. METHODS We trained rats in a delay-discounting task. Following acquisition of stable performance, rats were exposed to cocaine (10 mg/kg, intraperitoneal) every other day for 14 days and locomotor activity was measured. Two days later, delay-discounting performance was re-evaluated. To assess reversal of impulsivity, injections of a CB1R antagonist (1.5 mg/kg, intraperitoneal) or vehicle were given 30 minutes before the task. During the second experiment, aimed at preventing impulsivity rather than reversing it, CB1Rs were antagonized before each cocaine injection. In this experiment, subsecond dopamine release was measured in the nucleus accumbens during delay-discounting sessions before and after cocaine treatment. RESULTS Blockade of CB1Rs reversed and prevented cocaine-induced impulsivity. Electrochemical results showed that during baseline and following disruption of endocannabinoid signaling, there was a robust increase in dopamine for immediate large rewards compared with immediate small rewards, but this effect reversed when the delay for the large reward was 10 seconds. In contrast, dopamine release always increased for one-pellet options at minimal or moderate delays in vehicle-treated rats. CONCLUSIONS Endocannabinoids play a critical role in changes associated with cocaine exposure. Cannabinoid type 1 receptor blockade may thus counteract maladaptive alterations in afferents to dopamine neurons, thereby preventing changes in dopaminergic activity underlying a loss of self-control.

Dynamic changes in dopamine tone during self-stimulation of the ventral tegmental area in rats

In a prior study, phasic release of dopamine (DA) in the nucleus accumbens (NAc) was only transiently and rarely detected by means of fast-scan cyclic voltammetry (FCSV) in rats already trained to work for electrical stimulation of the ventral tegmental area (VTA) on a continuous reinforcement schedule. However, in rats receiving rewarding electrical stimulation via lateral hypothalamic (LH) electrodes, elevated DA tone in the NAc terminal field was detected via microdialysis for up to 2h, even when short (1.5s) inter-train intervals were employed. To better characterize the similarities and differences between the FSCV and microdialysis measurements, we trained rats to self-administer VTA stimulation under conditions similar to those employed in the initial FSCV study. The results resemble those obtained by means of microdialysis in rats receiving LH stimulation but differed from the prior FSCV data. Although the concentration of DA in dialysate obtained from NAc probes did fall after having peaked at the 30 min mark, this decline set in much later than in the FSCV studies, and elevated DA tone could still be detected after 110 min of self-stimulation. The stimulation-induced peak in DA tone could be restored by a 30 min rest period, a manipulation that was ineffective previously in restoring the FSCV measure of phasic release. These findings are discussed in terms of the differential sensitivity of the FSCV and microdialysis methods to phasic and tonic signaling by DA neurons and to different transitions between their activity states.

Role of Dopamine Tone in the Pursuit of Brain Stimulation Reward

Dopaminergic neurons contribute to intracranial self-stimulation (ICSS) and other reward-seeking behaviors, but it is not yet known where dopaminergic neurons intervene in the neural circuitry underlying reward pursuit or which psychological processes are involved. In rats working for electrical stimulation of the medial forebrain bundle, we assessed the effect of GBR-12909 (1-[2-[bis(4-fluorophenyl)-methoxy]ethyl]-4-[3- phenylpropyl]piperazine), a specific blocker of the dopamine transporter. Operant performance was measured as a function of the strength and cost of electrical stimulation. GBR-12909 increased the opportunity cost most subjects were willing to pay for a reward of a given intensity. However, this effect was smaller than that produced by a regimen of cocaine administration that drove similar increases in nucleus accumbens (NAc) dopamine levels in unstimulated rats. Delivery of rewarding stimulation to drug-treated rats caused an additional increase in dopamine concentration in the NAc shell in cocaine-treated, but not GBR-12909-treated, rats. These behavioral and neurochemical differences may reflect blockade of the norepinephrine transporter by cocaine but not by GBR-12909. Whereas the effect of psychomotor stimulants on ICSS has long been attributed to dopaminergic action at early stages of the reward pathway, the results reported here imply that increased dopamine tone boosts reward pursuit by acting at or beyond the output of the circuitry that temporally and spatially summates the output of the directly stimulated neurons underlying ICSS. The observed enhancement of reward seeking could be attributable to a decrease in the value of competing behaviors, a decrease in subjective effort costs, or an increase in reward-system gain.

Cannabinoid Receptor Blockade Reduces the Opportunity Cost at Which Rats Maintain Operant Performance for Rewarding Brain Stimulation

There is ample evidence that blockade of CB1 receptors reduces reward seeking. However, the reported effects of CB1 blockade on performance for rewarding electrical brain stimulation stand out as an exception. By applying a novel method for conceptualizing and measuring reward seeking, we show that AM-251, a CB1 receptor antagonist, does indeed decrease performance for rewarding electrical stimulation of the medial forebrain bundle in rats. Reward seeking depends on multiple sets of variables, including the intensity of the reward, its cost, and the value of competing rewards. In turn, reward intensity depends both on the sensitivity and gain of brain reward circuitry. We show that drug-induced changes in sensitivity cannot account for the suppressive effect of AM-251 on reward seeking. Therefore, the role of CB1 receptors must be sought among the remaining determinants of performance. Our analysis provides an explanation of the inconsistencies between prior reports, which likely arose from the following: (1) the averaging of data across subjects showing heterogeneous effects and (2) the use of methods that cannot distinguish between the different determinants of reward pursuit. By means of microdialysis, we demonstrate that blockade of CB1 receptors attenuates nucleus accumbens dopamine release in response to rewarding medial forebrain bundle stimulation, and we propose that this action is responsible for the ability of the drug to decrease performance for the electrical reward.

Predictable and unpredictable rewards produce similar changes in dopamine tone.

Unpredicted rewards trigger more vigorous phasic responses in midbrain dopamine (DA) neurons than predicted rewards. However, recent evidence suggests that reward predictability may fail to influence DA signaling over longer scales: In rats passively receiving rewarding electrical brain stimulation, the concentration of DA in dialysate obtained from nucleus accumbens probes was similar regardless of whether reward onset was predictable (G. Hernandez et al., 2006). The present experiment followed up on these findings by requiring the rats to work for the rewarding stimulation, thus confirming whether they indeed learned the timing and predictability of reward delivery. Performance under fixed-interval and variable-interval schedules was compared, and DA levels in the nucleus accumbens were measured by means of in vivo microdialysis. The observed patterns of operant responding indicate that the rats working under the fixed-interval schedule learned to predict the time of reward availability, whereas the rats working under the variable-interval schedule did not. Nonetheless, indistinguishable changes in DA concentration were observed in the 2 groups. Thus, reward predictability had no discernable effect on a measure believed to track the slower components of DA signaling.

Effect of CB1 Receptor Blockade on Food-Reinforced Responding and Associated Nucleus Accumbens Neuronal Activity in Rats

Studies have shown that disruption of cannabinoid receptor signaling reduces operant responses for rewards; yet it is unknown whether changes in neural activity at dopamine terminal regions such as the nucleus accumbens (NAc) underlie these behavioral effects. To study the neural correlates that accompany the disruption of endogenous cannabinoid (eCB) signaling in a food-motivated task, we recorded the neural activity and local field potentials (LFPs) from the NAc. A within-subject design was used for recordings as rats engaged in lever-pressing behavior for sucrose chocolate-flavored pellets delivered during responding in a progressive ratio (PR) schedule of reinforcement. Rats were food restricted to 85 ± 5% of their free body weight and trained under a PR until a stable breakpoint was observed (12 sessions ± 3). Once performance was stable, recordings were made under baseline, vehicle, and following administration of the cannabinoid inverse agonist rimonabant (150 μg/kg, i.v). NAc neurons encoded reward-predictive cues as well as food reward delivery. Rimonabant administration robustly reduced breakpoints in all rats tested, as previously reported. We found that this reduction is accompanied by a profound attenuation in the strength and coordination of specific event-related spiking activity. Moreover, rimonabant decreased LFP gamma power at 80 Hz (high gamma) at reward delivery and gamma power at 50 Hz (low gamma) at cue onset. Together the present results indicate that the eCB system sculpts neural activity patterns that accompany PR performance and reward consumption.

Dopamine tone increases similarly during predictable and unpredictable administration of rewarding brain stimulation at short inter-train intervals.

Unpredicted rewards, but not predicted ones, trigger strong phasic changes in the firing rates of midbrain dopamine (DA). In contrast, neurochemical measurements of DA tone have failed to reveal an influence of reward predictability. However, the subjects of the neurochemical experiments were asked to predict reward onset over longer intervals (12s, on average) than the subjects of the electrophysiological studies (typically, 2s). Thus, the contrasting effects of reward predictability could reflect the difference in the duration of the interval separating the predictor from the reward rather than a difference in the influence of reward predictability on phasic and tonic DA signaling. This hypothesis was tested in rats receiving trains of rewarding electrical brain stimulation with either a predictable or unpredictable onset. The mean inter-train interval was 1.5s, a value close to the 2-s CS-US interval that has been used in electrophysiological studies demonstrating the dependence of phasic DA responses on reward predictability. Despite the shortened inter-train interval, the time courses of the observed stimulation-induced elevations in DA levels were very similar, regardless of whether train onset was predictable. This finding is consistent with the idea that tonic DA signaling is insensitive to the predictability of rewards.

Potentiation of intracranial self-stimulation during prolonged subcutaneous infusion of cocaine

Subcutaneous administration of cocaine yields a longer duration of action than administration via the intraperitoneal or intravenous routes. However, cocaine is a powerful vasoconstrictor, and thus injection of this drug at a single subcutaneous locus entails significant risk of necrotic skin lesions. This paper introduces a new method for subcutaneous administration of cocaine that reduces the probability of dermonecrosis by dispersing the drug under a large area of skin. Two experiments were conducted to evaluate the new method. In the first, changes in dopamine tone in the nucleus accumbens were measured by means of microdialysis during prolonged subcutaneous infusions of cocaine. The dopamine concentration attained a fairly stable, elevated level, suggesting that absorption, distribution, and excretion of the drug approached steady state. In a second experiment, performance for rewarding electrical stimulation was measured during repeated prolonged infusions of cocaine. The pulse frequency required to sustain responding was decreased by the drug, in a manner that was stable both within and across test sessions. Thus, the new method is appropriate for studies requiring stable neurochemical and behavioral conditions during repeated long test sessions, high rates of drug delivery and alternation between administration of the drug and the vehicle.