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Repeated intermittent exposure to cocaine results in the neurochemical sensitization of dopamine (DA) transmission within the nucleus accumbens (NAc). Indeed, the excitability of DA neurons in the ventral tegmental area (VTA) is enhanced within hours of initial psychostimulant exposure. However, it is not known if this is accompanied by a comparably rapid change in the ability of cocaine to increase extracellular DA concentrations in the ventral striatum. To address this question we used fast-scan cyclic voltammetry (FSCV) in awake-behaving rats to measure DA responses in the NAc shell following an initial intravenous cocaine injection, and then again 2-hours later. Both injections quickly elevated DA levels in the NAc shell, but the second cocaine infusion produced a greater effect than the first, indicating sensitization. This suggests that a single injection of cocaine induces sensitization-related plasticity very rapidly within the mesolimbic DA system.

Though the last decade has seen accelerated advances in techniques and technologies to perturb neuronal circuitry in the brain, we are still poorly equipped to adequately dissect endogenous peptide release in vivo. To this end we developed a system that combines in vivo optogenetics with microdialysis and a highly sensitive mass spectrometry-based assay to measure opioid peptide release in freely moving rodents.

Dopamine cell firing can encode errors in reward prediction, providing a learning signal to guide future behavior. Yet dopamine is also a key modulator of motivation, invigorating current behavior. Existing theories propose that fast (“phasic”) dopamine fluctuations support learning, while much slower (“tonic”) dopamine changes are involved in motivation. We examined dopamine release in the nucleus accumbens across multiple time scales, using complementary microdialysis and voltammetric methods during adaptive decision-making. We first show that minute-by-minute dopamine levels covary with reward rate and motivational vigor. We then show that second-by-second dopamine release encodes an estimate of temporally-discounted future reward (a value function). We demonstrate that changing dopamine immediately alters willingness to work, and reinforces preceding action choices by encoding temporal-difference reward prediction errors. Our results indicate that dopamine conveys a single, rapidly-evolving decision variable, the available reward for investment of effort, that is employed for both learning and motivational functions.

While most drugs of abuse increase dopamine neurotransmission, rapid neurochemical measurements show that different drugs evoke distinct dopamine release patterns within the nucleus accumbens. Rapid changes in dopamine concentration following psychostimulant administration have been well studied; however, such changes have never been examined following opioid delivery. Here, we provide novel measures of rapid dopamine release following intravenous infusion of two opioids, morphine and oxycodone, in drug‐naïve rats using fast‐scan cyclic voltammetry and rapid (1 min) microdialysis coupled with high‐performance liquid chromatography ‐ tandem mass spectrometry (HPLC‐MS). In addition to measuring rapid dopamine transmission, microdialysis HPLC‐MS measures changes in GABA, glutamate, monoamines, monoamine metabolites and several other neurotransmitters. Although both opioids increased dopamine release in the nucleus accumbens, their patterns of drug‐evoked dopamine transmission differed dramatically. Oxycodone evoked a robust and stable increase in dopamine concentration and a robust increase in the frequency and amplitude of phasic dopamine release events. Conversely, morphine evoked a brief (~ 1 min) increase in dopamine that was coincident with a surge in GABA concentration and then both transmitters returned to baseline levels. Thus, by providing rapid measures of neurotransmission, this study reveals previously unknown differences in opioid‐induced neurotransmitter signaling. Investigating these differences may be essential for understanding how these two drugs of abuse could differentially usurp motivational circuitry and powerfully influence behavior.

Prairie voles (Microtus ochrogaster) are a socially monogamous rodent species and their cooperative behaviors require extensive communication between conspecifics. Rodents use ultrasonic vocalizations (USVs) to communicate and because a prairie vole breeder pair must engage in extensive cooperation for successful reproduction, auditory communication may be critical for this species. Therefore, we sought to characterize USVs in adult male and female prairie voles, and to determine how these calls are influenced by social context, salient social stimuli and the psychostimulant drug of abuse amphetamine (AMPH). Here, we characterize prairie vole USVs by showing the range of frequencies of prairie vole USVs, the proportion of various call types, how these call types compare between males and females, and how they are influenced by social stimulation and AMPH. AMPH caused a robust increase in the number of USVs in both males and females and there was a dramatic sex difference in the complexity of call structures of AMPH-induced USVs, with males emitting more elaborate calls. Moreover, we show that novel (i.e. salient) social cues evoked differential increases in USVs across sex, with males showing a much more robust increase in USV production, both with respect to the frequency and complexity of USV production. Exposure to an estrous female in particular caused an extraordinary increase in USVs in male subjects. These data suggest that USVs may be a useful measure of social motivation in this species, including how social behaviors can be impacted by drugs of abuse.

To survive in a dynamic environment, animals must identify changes in resource availability and rapidly apply adaptive strategies to obtain resources that promote survival. We have utilised a behavioral paradigm to assess differences in foraging strategy when resource (reward) availability unexpectedly changes. When reward magnitude was reduced by 50% (receive one reward pellet instead of two), male and female rats developed a preference for the optimal choice by the second session. However, when an expected reward was omitted (receive no reward pellets instead of one), subjects displayed a robust preference for the optimal choice during the very first session. Previous research shows that, when an expected reward is omitted, dopamine neurons phasically decrease their firing rate, which is hypothesised to decrease dopamine release preferentially affecting D2-like receptors. As robust changes in behavioral preference were specific to reward omission, we tested this hypothesis and the functional role of D1- and D2-like receptors in the nucleus accumbens in mediating the rapid development of a behavioral preference for the rewarded option during reward omission in male rats. Blockade of both receptor types had no effect on this behavior; however, holding D2-like, but not D1-like, receptor tone via infusion of dopamine receptor agonists prevented the development of the preference for the rewarded option during reward omission. These results demonstrate that avoiding an outcome that has been tagged with aversive motivational properties is facilitated through decreased dopamine transmission and subsequent functional disruption of D2-like, but not D1-like, receptor tone in the nucleus accumbens.

The sensory properties of a reward-paired cue (a conditioned stimulus; CS) may impact the motivational value attributed to the cue, and in turn influence the form of the conditioned response (CR) that develops. A cue with multiple sensory qualities, such as a moving lever-CS, may activate numerous neural pathways that process auditory and visual information, resulting in CRs that vary both within and between individuals. For example, CRs include approach to the lever-CS itself (rats that “sign-track”; ST), approach to the location of reward delivery (rats that “goal-track”; GT), or an “intermediate” combination of these behaviors. We found that the multimodal sensory features of the lever-CS were important to the development and expression of sign-tracking. When the lever-CS was covered, and thus could only be heard moving, STs not only continued to approach the lever location but also started to approach the food cup during the CS period. While still predictive of reward, the auditory component of the lever-CS was a much weaker conditioned reinforcer than the visible lever-CS. This plasticity in behavioral responding observed in STs closely resembled behaviors normally seen in rats classified as “intermediates.” Furthermore, the ability of both the lever-CS and the reward-delivery to evoke dopamine release in the nucleus accumbens was also altered by covering the lever—dopamine signaling in STs resembled neurotransmission observed in rats that normally only GT. These data suggest that while the visible lever-CS was attractive, wanted, and had incentive value for STs, when presented in isolation, the auditory component of the cue was simply predictive of reward, lacking incentive salience. Therefore, the specific sensory features of cues may differentially contribute to responding and ensure behavioral flexibility.

Although populations of neurons are known to vary on the micrometer scale, little is known about whether basal concentrations of neurotransmitters also vary on this scale. We used low-flow push-pull perfusion to test if such chemical gradients exist between several small brain nuclei. A miniaturized polyimide-encased push-pull probe was developed and used to measure basal neurotransmitter spatial gradients within brain of live animals with 0.004 mm(3) resolution. We simultaneously measured dopamine (DA), norepinephrine, serotonin (5-HT), glutamate, γ-aminobutyric acid (GABA), aspartate (Asp), glycine (Gly), acetylcholine (ACh), and several neurotransmitter metabolites. Significant differences in basal concentrations between midbrain regions as little as 200 μm apart were observed. For example, dopamine in the ventral tegmental area (VTA) was 4.8 ± 1.5 nM but in the red nucleus was 0.5 ± 0.2 nM. Regions of high glutamate concentration and variability were found within the VTA of some individuals, suggesting hot spots of glutamatergic activity. Measurements were also made within the nucleus accumbens core and shell. Differences were not observed in dopamine and 5-HT in the core and shell; but their metabolites homovanillic acid (460 ± 60 nM and 130 ± 60 nM respectively) and 5-hydroxyindoleacetic acid (720 ± 200 nM and 220 ± 50 nM respectively) did differ significantly, suggesting differences in dopamine and 5-HT activity in these brain regions. Maintenance of these gradients depends upon a variety of mechanisms. Such gradients likely underlie highly localized effects of drugs and control of behavior that have been found using other techniques.

In addition to blocking dopamine (DA) uptake, cocaine also causes an unconditioned increase in DA release. In drug naive rats, this effect is most robust within the nucleus accumbens (NAc) shell. Recent studies have shown that, in rats trained to self-administer cocaine, cocaine may act in the periphery to enhance mesolimbic DA release. Further, these studies have suggested that peripheral cocaine action may also enhance unconditioned DA release. Here, we test if it is necessary for cocaine to enter the brain to evoke unconditioned increases in DA release within the NAc shell. Administration of a cocaine analogue that crosses the blood brain barrier (cocaine HCl) enhances electrically evoked DA release and the number of cocaine-evoked phasic DA release events (i.e., DA transients) within the NAc shell. However, administration of a cocaine analogue that does not cross the blood brain barrier (cocaine MI) does not alter either measure. We therefore conclude that cocaine must act within the central nervous system to evoke unconditioned DA release within the NAc shell.

Background Prominent neurobiological theories of addiction posit a central role for aberrant mesolimbic dopamine release but disagree as to whether repeated drug experience blunts or enhances this system. Although drug withdrawal diminishes dopamine release, drug sensitization augments mesolimbic function, and both processes have been linked to drug seeking. One possibility is that the dopamine system can rapidly switch from dampened to enhanced release depending on the specific drug-predictive environment. To test this, we examined dopamine release when cues signaled delayed cocaine delivery versus imminent cocaine self-administration. Methods Fast-scan cyclic voltammetry was used to examine real-time dopamine release while simultaneously monitoring behavioral indexes of aversion as rats experienced a sweet taste cue that predicted delayed cocaine availability and during self-administration. Furthermore, the impact of cues signaling delayed drug availability on intracranial self-stimulation, a broad measure of reward function, was assessed. Results We observed decreased mesolimbic dopamine concentrations, decreased reward sensitivity, and negative affect in response to the cocaine-predictive taste cue that signaled delayed cocaine availability. Importantly, dopamine concentration rapidly switched to elevated levels to cues signaling imminent cocaine delivery in the subsequent self-administration session. Conclusions These findings show rapid, bivalent contextual control over brain reward processing, affect, and motivated behavior and have implications for mechanisms mediating substance abuse.

Background Dopamine signaling in the nucleus accumbens (NAc) is essential for goal-directed behaviors and primarily arises from burst firing of ventral tegmental area neurons. However, the role of associative neural substrates such as the basolateral amygdala (BLA) in regulating phasic dopamine release in the NAc, particularly during reward seeking, remains unknown. Methods Male Sprague-Dawley rats learned to discriminate two cues: a discriminative stimulus (DS) that predicted sucrose reinforcement contingent upon a lever press and a nonassociated stimulus (NS) that predicted a second lever never reinforced with sucrose. Following training, a test session was completed in which NAc dopamine was measured using fast-scan cyclic voltammetry in conjunction with inactivation of the ipsilateral BLA (gamma-aminobutyric acid agonists; baclofen/muscimol) to determine the contribution of BLA activity to dopamine release in the NAc core during the task. Results Under vehicle conditions, DS and NS presentation elicited dopamine release within the NAc core. The DS evoked significantly more dopamine than the NS. Inactivation of the BLA selectively attenuated the magnitude of DS-evoked dopamine release, concurrent with an attenuation of DS-evoked conditioned approaches. Other behavioral responses (e.g., lever pressing) and dopamine release concomitant with those events were unaltered by BLA inactivation. Furthermore, neither ventral tegmental area electrically stimulated dopamine release nor the probability of high concentration dopamine release events was altered following BLA inactivation. Conclusions These results demonstrate that the BLA terminally modulates dopamine signals within the NAc core under specific, behaviorally relevant conditions, illustrating a functional mechanism by which the BLA selectively facilitates responding to motivationally salient environmental stimuli.

The prairie vole ( Microtus ochrogaster ) is a socially monogamous rodent species that forms pair bonds after mating, a behavior in which central dopamine (DA) has been implicated. Here, we used male prairie voles to examine the effects of drug exposure on pair bonding and related neural circuitry. In our first experiment, amphetamine (AMPH) motivated behavior was examined using a conditioned place preference (CPP) paradigm and was shown to be mediated by activation of D1-like DA receptors. Next, we examined the effects of repeated AMPH exposure on pair bonding. Intact and saline pretreated control males displayed mating-induced partner preferences, whereas males pretreated with AMPH at the doses effective to induce CPP failed to show mating-induced partner preferences. Such AMPH treatment also enhanced D1, but not D2, DA receptor expression in the nucleus accumbens (NAcc). Furthermore, pharmacological blockade of D1-like DA receptors in the NAcc rescued mating-induced partner preferences in AMPH-treated males. Together, our data indicate that repeated AMPH exposure may narrow the behavioral repertoire of male prairie voles via a DA receptor-specific mechanism in the NAcc, resulting in the impairment of pair bond formation.

Drug seeking is significantly regulated by drug-associated cues and associative learning between environmental cues and cocaine reward is mediated by dopamine transmission within the nucleus accumbens (NAc). However, dopamine transmission during early acquisition of a cue-cocaine association has never been assessed because of the technical difficulties associated with resolving cue-evoked and cocaine-evoked dopamine release within the same conditioning trial. Here, we used fast-scan cyclic voltammetry to measure sub-second fluctuations in dopamine concentration within the NAc core and shell during the initial acquisition of a cue-cocaine Pavlovian association. Within the NAc core, cue-evoked dopamine release developed during conditioning. However, within the NAc shell, the predictive cue appeared to cause an unconditioned decrease in dopamine concentration. The pharmacological effects of cocaine also differed between sub-regions, as cocaine increased phasic dopamine release events within the NAc shell but not the core. Thus, real-time measurements not only revealed the initial development of a conditioned neurochemical response but also demonstrated differential phasic dopamine transmission patterns across NAc sub-regions during the acquisition of a cue-cocaine association.

Preferential enhancement of dopamine transmission within the nucleus accumbens (NAc) shell is a fundamental aspect of the neural regulation of cocaine reward. Despite its importance, the nature of this effect is poorly understood. Here, we used fast-scan cyclic voltammetry to examine specific transmission processes underlying cocaine-evoked increases in dopamine transmission within the NAc core and shell. Initially, we examined altered terminal dopamine concentrations after global autoreceptor blockade. This was the first examination of autoreceptor regulation of naturally occurring phasic dopamine transmission and provided a novel characterization of specific components of dopamine neurotransmission. Comparison of increased dopamine signaling evoked by autoreceptor blockade and cocaine administration allowed robust resolution between increased frequency, concentration, and duration of phasic dopamine release events after cocaine delivery. Cocaine increased dopamine transmission by slowed uptake and increased concentration of dopamine released in the core and shell. However, an additional increase in the number phasic release events occurred only within the NAc shell, and this increase was eliminated by inactivation of midbrain dopaminergic neurons. This represents the first evidence that cocaine directly increases the frequency of dopamine release events and reveals that this is responsible for preferentially increased dopamine transmission within the NAc shell after cocaine administration. Additionally, cocaine administration resulted in a synergistic increase in dopamine concentration, and subregion differences were abolished when cocaine was administered in the absence of autoregulation. Together, these results demonstrate that cocaine administration results in a temporally and regionally specific increase in phasic dopamine release that is significantly regulated by dopamine autoreceptors.

Prairie vole breeder pairs form monogamous pair bonds, which are maintained through the expression of selective aggression toward novel conspecifics. Here, we utilize behavioral and anatomical techniques to extend the current understanding of neural mechanisms that mediate pair bond maintenance. For both sexes, we show that pair bonding up-regulates mRNA expression for genes encoding D1-like dopamine (DA) receptors and dynorphin as well as enhances stimulated DA release within the nucleus accumbens (NAc). We next show that D1-like receptor regulation of selective aggression is mediated through downstream activation of kappa-opioid receptors (KORs) and that activation of these receptors mediates social avoidance. Finally, we also identified sex-specific alterations in KOR binding density within the NAc shell of paired males and demonstrate that this alteration contributes to the neuroprotective effect of pair bonding against drug reward. Together, these findings suggest motivational and valence processing systems interact to mediate the maintenance of social bonds. DOI: http://dx.doi.org/10.7554/eLife.15325.001, eLife digest The bond between parents is one of the most important social relationships that humans have. Prairie voles are one of the few other mammals whose individuals also form long-term social bonds after having offspring together, so they have frequently been used to study the brain mechanisms that underlie such bonding. However, most previous studies have focused only on how the bond between a pair of mating partners is formed: little is known about how this bond is then maintained over months and years. When a prairie vole forms a bond with a mate, it will then aggressively reject other prairie voles. This “selective aggression” only happens once a social bond between two mating prairie voles is formed, so this behavior can be used as a proxy to confirm that the social bond exists. In order to study how prairie voles maintain bonds with a mate, Resendez et al. tracked what happens in the brain of a prairie vole during selective aggression. The experiments showed that this aggressive behaviour coincides with changes in gene expression and brain chemistry that make it unpleasant for a prairie vole to be exposed to voles that are not its partner. For male prairie voles – but not females – these changes only happened if the female mating partner became pregnant during the cohabitation period. The changes that occur in the brain as a result of bonding with a partner also mean that drugs that are normally addictive are no longer pleasant and rewarding to the prairie vole. Indeed, forming a social bond between mating animals alters the brain in similar ways to the effects produced by addictive drugs. Thus, in a sense, each member of the mating pair becomes ‘addicted’ to their partner. The results presented by Resendez et al. also have implications for humans. They suggest that having a strong social support network is a powerful way of preventing casual drug use from developing into compulsive drug addiction. This may also mean that positive social relationships could help to treat people with drug addiction problems. DOI: http://dx.doi.org/10.7554/eLife.15325.002

The formation of monogamous pair bonds, by prairie voles, is facilitated by activation of dopamine (DA) D2-like, but not D1-like, receptors within the nucleus accumbens (NAcc) shell. Because DA exerts opposing regulation of cAMP production depending on the subtype of receptor activated, we tested the hypothesis that DA regulation of pair bond formation is mediated via the cAMP signaling cascade. Consistent with activation of D2-like receptors, decreasing cAMP signaling, by blocking cAMP binding sites on protein kinase A (PKA), facilitated partner preference formation. Conversely, increasing cAMP signaling, by preventing the activation of inhibitory G-proteins, activating stimulatory G-proteins, or stimulating PKA prevented the formation of mating-induced partner preferences. These manipulations were effective in the shell, but not the core, of the NAcc. Together, these data demonstrate opposing regulation over pair bond formation by cAMP signaling within the NAcc shell.

SummaryIntracranial self-stimulation (ICSS) activates the neural pathways that mediate reward, including dopaminergic terminal areas such as the nucleus accumbens (NAc). However, a direct role of dopamine in ICSS-mediated reward has been questioned. Here, simultaneous voltammetric and electrophysiological recordings from the same electrode reveal that, at certain sites, the onset of anticipatory dopamine surges and changes in neuronal firing patterns during ICSS are coincident, whereas sites lacking dopamine changes also lack patterned firing. Intrashell microinfusion of a D1, but not a D2 receptor antagonist, blocks ICSS. An iontophoresis approach was implemented to explore the effect of dopamine antagonists on firing patterns without altering behavior. Similar to the microinfusion experiments, ICSS-related firing is selectively attenuated following D1 receptor blockade. This work establishes a temporal link between anticipatory rises of dopamine and firing patterns in the NAc shell during ICSS and suggests that they may play a similar role with natural rewards and during drug self-administration.

Transient surges of dopamine in the nucleus accumbens are associated with drug seeking. Using a voltammetric sensor with high temporal and spatial resolution, we demonstrate differences in the temporal profile of dopamine concentration transients caused by acute doses of nicotine, ethanol, and cocaine in the nucleus accumbens shell of freely moving rats. Despite differential release dynamics, all drug effects are uniformly inhibited by administration of rimonabant, a cannabinoid receptor (CB1) antagonist, suggesting that an increase in endocannabinoid tone facilitates the effects of commonly abused drugs on subsecond dopamine release. These time-resolved chemical measurements provide unique insight into the neurobiological effectiveness of rimonabant in treating addictive disorders.

Pair bonding represents social attachment between mates and is common among monogamous animals. The prairie vole (Microtus ochrogaster) is a monogamous rodent in which mating facilitates pair bond formation. In this review, we first discuss how prairie voles have been used as an excellent model for neurobiological studies of pair bonding. We then primarily focus on male prairie voles to summarize recent findings from neuroanatomical, neurochemical, cellular, molecular, and behavioral studies implicating vasopressin (AVP), oxytocin (OT), and dopamine (DA) in the regulation of pair bonding. Possible interactions among these neurochemicals in the regulation of pair bonding, the brain areas important for pair bond formation, and potential sexually dimorphic mechanisms underlying pair bonding are also discussed. As analogous social bonds are formed by humans, investigation of the neurochemical regulation of pair bond formation in prairie voles may be beneficial for our understanding of the mechanisms associated with normal and abnormal social behaviors in humans.

Pair bond formation has been investigated much less than many other social behaviors, perhaps in part because traditional laboratory mice and rats do not exhibit this behavior. However, pair bonding is common among monogamous animals such as the prairie vole (Microtus ochrogaster). In this review, we discuss how the prairie vole has been used as a model system to investigate the neurobiology of pair bonding. Descriptions include neuroanatomical differences between monogamous and non-monogamous voles, as well as how manipulations of vasopressin, oxytocin, dopamine, and corticosterone systems affect pair bond formation. Also summarized are potential interactions among these systems that regulate pair bonding, and the extent of sexual dimorphism in underlying mechanisms. Pair bonding in prairie voles is an excellent model system for studying central processing of social information. Understanding the mechanisms underlying this behavior may provide important insights into human disorders associated with impaired social functioning.

Rats anticipate daily meals with increased approaches to a feeder and an increase in core body temperature. Food-anticipatory activity (FAA) is thought to be under the control of a feeding-entrained circadian oscillator (FEO). Ibotenic acid and electrolytic lesions in the region of the parabrachial nuclei (PBN) in the rat severely disrupt FAA (feeder approaches) and temperature rhythms. The PBN receive dense input from the area postrema (AP), which lacks a blood-brain barrier and thus has access to humoral factors in the systemic circulation. The present study assesses development and maintenance of FAA in rats with cautery lesions of the AP. The results demonstrate that AP lesions do not alter FAA. This experiment does not support the hypothesis that the AP in the caudal brainstem detects and relays circulating signals from the periphery that trigger FAA.

The prairie vole is a socially monogamous rodent that is an excellent animal model for studies of the neurobiology of social attachment. Such studies have demonstrated that activation of reward circuitry during social interactions facilitates pair bond formation. Within this circuitry, μ-opioid receptors (MORs) modulate naturally rewarding behavior in an anatomically segregated manner; MORs located throughout the striatum (dorsal striatum, NAc core, and the entire NAc shell) are implicated in general motivational processes, whereas those located specifically within the dorsomedial NAc shell mediate positive hedonics (and are referred to as a “hedonic hotspot”). The purpose of the present study was to determine whether MORs within these distinct subregions differentially mediate pair bond formation. We first used receptor autoradiography to compare MOR binding densities between these regions. MOR binding was significantly higher in the NAc core and dorsomedial NAc shell compared with the ventral NAc shell. We next used partner preference testing to determine whether MORs within these subregions differentially mediate pair bonding. Blockade of MORs using 1 or 3 μg of H-d-Phe-Cys-Tyr-d-Trp-Arg-Thr-Pen-Thr-NH2within the dorsal striatum decreased mating during the cohabitation period and inhibited partner preference formation. In contrast, blockade of MORs within dorsomedial NAc shell inhibited partner preference formation without effecting mating behavior, whereas other regions were not involved. Thus, MORs within the dorsal striatum mediate partner preference formation via impairment of mating, whereas those in the dorsomedial NAc shell appear to mediate pair bond formation through the positive hedonics associated with mating.

Although fear directs adaptive behavioral responses, how aversive cues recruit motivational neural circuitry is poorly understood. Specifically, while it is known that dopamine (DA) transmission within the nucleus accumbens (NAc) is imperative for mediating appetitive motivated behaviors, its role in aversive behavior is controversial. It has been proposed that divergent phasic DA transmission following aversive events may correspond to segregated mesolimbic dopamine pathways; however, this prediction has never been tested. Here, we used fast-scan cyclic voltammetry to examine real-time DA transmission within NAc core and shell projection systems in response to a fear-evoking cue. In male Sprague Dawley rats, we first demonstrate that a fear cue results in decreased DA transmission within the NAc core, but increased transmission within the NAc shell. We examined whether these changes in DA transmission could be attributed to modulation of phasic transmission evoked by cue presentation. We found that cue presentation decreased the probability of phasic DA release in the core, while the same cue enhanced the amplitude of release events in the NAc shell. We further characterized the relationship between freezing and both changes in DA as well as local pH. Although we found that both analytes were significantly correlated with freezing in the NAc across the session, changes in DA were not strictly associated with freezing while basic pH shifts in the core more consistently followed behavioral expression. Together, these results provide the first real-time neurochemical evidence that aversive cues differentially modulate distinct DA projection systems.

Social bonds are important for human health and well-being, and a crucial component of these bonds is the ability to maintain a bond once it has been formed. Importantly, although bond maintenance is required for social attachments, very little is known about the neural mechanisms that mediate this behavior. Recently, laboratory studies utilizing the socially monogamous prairie vole (an excellent animal model for the neurobiology of selective social attachment), have allowed the neural correlates of selective social attachment to begin to unfold. These studies have identified that the activation of both motivational and hedonic processing systems, which mediate other natural rewards, is also important for mediating social behaviors that are characteristic of an established pair bond. These social behaviors include appetitive and positive social interactions with a potential mating partner in sexually naive prairie voles, the avoidance of novel conspecifics (and sometimes aggressive rejection) that characterizes the established pair bond and, finally, an aversion towards partner separation. The following review will discuss how a balance between opposing endogenous opioid systems - positive (mu-opiod receptors) and aversive (kappa-opioid receptors) - provide essential hedonic signaling that guides socially motivated behaviors.

The prairie vole is a socially monogamous species in which breeder pairs typically show strong and selective pair bonds. The establishment of a pair bond is associated with a behavioral transition from general affiliation to aggressive rejection of novel conspecifics. This “selective aggression” is indicative of mate guarding that is necessary to maintain the initial pair bond. In the laboratory, the neurobiology of this behavior is studied using resident-intruder testing. Although it is well established that social behaviors in other species are mediated by endogenous opioid systems, opiate regulation of pair bond maintenance has never been studied. Here, we used resident-intruder testing to determine whether endogenous opioids within brain motivational circuitry mediate selective aggression in prairie voles. We first show that peripheral blockade of κ-opioid receptors with the antagonist norbinaltorphimine (nor-BNI; 100 mg/kg), but not with the preferential μ-opioid receptor antagonist naloxone (1, 10, or 30 mg/kg), decreased selective aggression in males. We then provide the first comprehensive characterization of κ- and μ-opioid receptors in the prairie vole brain. Finally, we demonstrate that blockade of κ-opioid receptors (500 ng nor-BNI) within the nucleus accumbens (NAc) shell abolishes selective aggression in both sexes, but blockade of these receptors within the NAc core enhances this behavior specifically in females. Blockade of κ-opioid receptors within the ventral pallidum or μ-opioid receptors with the specific μ-opioid receptor antagonist H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-PenThr-NH2 (1 ng CTAP) within the NAc shell had no effect in either sex. Thus, κ-opioid receptors within the NAc shell mediate aversive social motivation that is critical for pair bond maintenance.

In their recent Perspective article, Roy Wise and Eugene Kiyatkin conclude that, “the primary, unconditioned rewarding effects of cocaine in naive animals involves the blockade of dopamine reuptake rather than stimulation of dopamine release” (Differentiating the rapid actions of cocaine. Nature Rev. Neurosci. 12, 479–484 (2011))1. However, recent voltammetry studies that have measured real-time changes in dopamine concentration at the instant that cocaine enters the brain have reached different conclusions2, 3, 4, 5. These studies show that intravenous cocaine delivery inhibits dopamine uptake in less than 5 seconds following drug delivery4, 5 and increases the frequency of phasic dopamine release events within the nucleus accumbens (NAc) shell to a greater extent than in the NAc core2, 3, 6, 7. This shell-specific increase in dopamine release begins approximately 10 seconds after cocaine delivery and lasts for approximately 2 minutes2. Unconditioned cocaine-evoked dopamine release is therefore both temporally and regionally specific and is important because it mediates the primary rewarding effects of cocaine3, 8.

The monogamous prairie vole displays developmental sensitivity to early pharmacological manipulation in a number of species-typical social behaviors. The long-term consequences of altering the neonatal dopamine system are not well characterized. This study examined whether early manipulation of the dopamine system, a known mediator of adult prairie vole social behavior, during neonatal development would affect adult aggressive and attachment behaviors. Eight-day-old pups were given a single treatment with either 1 mg/kg of SKF38393 (D1 agonist), quinpirole (D2 agonist), SCH23390 (D1 antagonist), eticlopride (D2 antagonist), or saline vehicle. As adults, animals received tests for intrasexual aggression and partner preference. Activation of D1-like receptors in pups impaired partner preference formation, but had no effect on aggression. Other neonatal treatments had no effect on their behavior as adults. To determine whether D1 activation in pups induced changes in dopamine receptor expression, we performed autoradiography on striatal tissue from a second cohort of saline-treated and SKF38393-treated animals. Although sex differences were observed, we found no treatment differences in D1 or D2 receptor binding in any striatal subregion. This study shows that exposure to a single early pharmacological alteration of dopamine receptor activity may have long-term effects on the social behavior of prairie voles. © 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins.

Although the protective effects of social bonds on drug use/abuse have been well documented, we know little about the underlying neural mechanisms. Using the prairie vole (Microtus ochrogaster)—a socially monogamous rodent that forms long-term pair bonds after mating—we demonstrate that amphetamine (AMPH) conditioning induced a conditioned place preference (CPP) in sexually naive (SN), but not pair-bonded (PB), males. Although AMPH treatment induced a similar magnitude of dopamine release in the nucleus accumbens (NAcc) of SN and PB males, it had differential effects on NAcc D1 receptor (D1R) binding. Specifically, AMPH treatment increased D1R binding in SN, but decreased D1R binding in PB males. NAcc D1R, but not D2 receptor, antagonism blocked AMPH-induced CPP in SN males and NAcc D1R activation before AMPH conditioning enabled AMPH-induced CPP in PB males. Together, our data demonstrate that pair-bonding experience decreases the rewarding properties of AMPH through a D1R-mediated mechanism.

Fast-scan cyclic voltammetry (FSCV) at carbon-fiber microelectrodes has been used to demonstrate that sub-second changes in catecholamine concentration occur within the nucleus accumbens (NAc) shell during motivated behaviors, and these fluctuations have been attributed to rapid dopamine signaling. However, FSCV cannot distinguish between dopamine and norepinephrine, and caudal regions of the NAc shell receive noradrenergic projections. Therefore, in the present study, we examined the degree to which norepinephrine contributes to catecholamine release within rostral and caudal portion of NAc shell. Analysis of tissue content revealed that dopamine was the major catecholamine detectable in the rostral NAc shell, whereas both dopamine and norepinephrine were found in the caudal subregion. To examine releasable catecholamines, electrical stimulation was used to evoke release in anesthetized rats with either stimulation of the medial forebrain bundle, a pathway containing both dopaminergic and noradrenergic projections to the NAc, or the ventral tegmental area/substantia nigra, the origin of dopaminergic projections. The catecholamines were distinguished by their responses to different pharmacological agents. The dopamine autoreceptor blocker, raclopride, as well as the monoamine and dopamine transporter blockers, cocaine and GBR 12909, increased evoked catecholamine overflow in both the rostral and caudal NAc shell. The norepinephrine autoreceptor blocker, yohimbine, and the norepinephrine transporter blocker, desipramine, increased catecholamine overflow in the caudal NAc shell without significant alteration of evoked responses in the rostral NAc shell. Thus, the neurochemical and pharmacological results show that norepinephrine signaling is restricted to caudal portions of the NAc shell. Following raclopride and cocaine or raclopride and GBR 12909, robust catecholamine transients were observed within the rostral shell but these were far less apparent in the caudal NAc shell, and they did not occur following yohimbine and desipramine. Taken together, the data demonstrate that catecholamine signals in the rostral NAc shell detected by FSCV are due to change in dopamine transmission.

Many individual neurons within the intact brain fire in stochastic patterns that arise from interactions with the neuronal circuits that they comprise. However, the chemical communication that is evoked by these firing patterns has not been characterized because sensors suitable to monitor subsecond chemical events in micron dimensions have only recently become available. Here we employ a voltammetric sensor technology coupled with principal component regression to examine the dynamics of dopamine concentrations in the nucleus accumbens (NAc) of awake and unrestrained rats. The sensor has submillimeter dimensions and provides high temporal (0.1 s) resolution. At select locations spontaneous dopamine transient concentration changes were detected, achieving instantaneous concentrations of approximately 50 nm. At other locations, transients were absent even though dopamine was available for release as shown by extracellular dopamine increases following electrical activation of dopaminergic neurons. At sites where dopamine concentration transients occur, uptake inhibition by cocaine enhances the frequency and magnitude of the rapid transients while also causing a more gradual increase in extracellular dopamine. These effects were largely absent from sites that did not support ongoing transient activity. These findings reveal an unanticipated spatial and temporal heterogeneity of dopamine transmission within the NAc that may depend upon the firing of specific subpopulations of dopamine neurons.

Recent studies have shown that the neural regulation of pair bonding in the monogamous prairie vole (Microtus ochrogaster) is similar to that of drug seeking in more traditional laboratory rodents. Therefore, strong interactions between social behavior and drug reward can be expected. Here, we established the prairie vole as a model for drug studies by demonstrating robust amphetamine-induced conditioned place preferences in this species. For both males and females, the effects of amphetamine were dose-dependent, with females being more sensitive to drug treatment. This study represents the first evidence of drug reward in this species. Future studies will examine the effects of social behavior on drug reward and the underlying neurobiology of such interactions.

The involvement of dopamine within the nucleus accumbens in the formation and maintenance of pair bonds was assessed in a series of experiments using the monogamous prairie vole. We show that dopamine transmission that promotes pair bond formation occurs within the rostral shell of the nucleus accumbens, but not in its core or caudal shell. Within this specific brain region, D1- and D2-like receptor activation produced opposite effects: D1-like activation prevented pair bond formation, whereas D2-like activation facilitated it. After extended cohabitation with a female, male voles showed behavior indicative of pair bond maintenance—namely, selective aggression towards unfamiliar females. These voles also showed a significant upregulation in nucleus accumbens D1-like receptors, and blockade of these receptors abolished selective aggression. Thus, neuroplastic reorganization of the nucleus accumbens dopamine system is responsible for the enduring nature of monogamous pair bonding. Finally, we show that this system may also contribute to species-specific social organization.

Comparisons between monogamous and promiscuous vole species have proven useful in examining neurobiological mechanisms underlying social attachment. Reward processing is important for social attachment, and the medial prefrontal cortex (mPFC) exerts a direct influence on reward pathways. Dopamine (DA), oxytocin (OT), and arginine vasopressin (AVP) all have been implicated in the regulation of social attachment in monogamous voles. Therefore, we used radiolabeled ligands to examine dopamine D1- and D2-like, OT, and AVP V1a receptor binding densities in the mPFC of monogamous and promiscuous voles. Species differences were found; monogamous voles had higher densities of D2-like and OT receptor binding and lower densities of D1-like and V1a receptor binding than did promiscuous voles. Sex differences also were found; females had higher densities of OT receptor binding but lower densities of V1a receptor binding than did males in both species. Further, the laminar distribution of receptor binding indicates the possibility of an interaction between DA and OT systems in the mPFC in the regulation of social attachment. Differences in D1- and D2-like receptor binding between species are discussed in terms of how they might modulate cortical activity and subsequent DA release in the nucleus accumbens (NAcc). © 2005 Elsevier Ireland Ltd. All rights reserved.

Although the role of nucleus accumbens (NAcc) dopamine (DA) in reward learning has been extensively studied, few investigations have addressed its involvement in learning socially relevant information. Here, we have examined the involvement of NAcc DA in social attachment of the "monogamous" prairie vole (Microtus orchrogaster). We first demonstrated that DA is necessary for the formation of social attachment in male prairie voles, because administration of haloperidol blocked, whereas apomorphine induced, partner-preference formation. We then provided the first descriptions of DA neuroanatomy and tissue content in vole NAcc, and mating appeared to induce a 33% increase in DA turnover. We also showed that administration of haloperidol directly into the NAcc blocked partner preferences induced by mating and apomorphine. In addition, administration of apomorphine into the NAcc but not the caudate putamen induced partner preferences in the absence of mating. Together, our data support the hypothesis that NAcc DA is critical for pair-bond formation in male prairie voles.

The ability to form an association between the time and the place of food availability, namely time-place learning, is presumably important for survival. The present study was designed toexamine time-place learning and to identify exogenous and endogenous factors that may affect this behavior in rats. In an initial experiment, rats displayed poor time-place behavior and appeared to prefer the feeder that was closer to the center aisle and water supply. When these cueswere minimized in a subsequent experiment, rats consistently displayed the time-place discrimination by exhibiting food anticipatory activity (FAA) at the correct location prior to each mealtime. These rats also showed significant correlations between the level of FAA and the amount of dopamine turnover (the dihydroxyphenylacetic acid/dopamine ratio) in the nucleus accumbens and paraventricular nucleus of the hypothalamus, indicating possible involvement of regional dopaminergic activity in time-place behavior. No correlation was found for norepinephrine, epinephrine, or serotonin. In addition, the correlation between FAA and dopamine turnover was not foundwhen rats were entrained to only one meal per day. Together, the data suggest that rats can learn the time- place discrimination under proper experimental conditions and that dopamine may play a role in the expression of this behavior.

Restricted daily food access acts as an entraining stimulus (zeitgeber) for a circadian clock, the feeding-entrainable oscillator (FEO). There are many properties of a daily meal that could potentially convey timing information to the FEO. Olfactory cues associated with feeding are one such property. In order to rule out olfaction as a necessary entraining stimulus, olfactory bulbectomized and sham-operated male Sprague–Dawley rats had access to food for 2 h each day. Food bin approach behavior was monitored as an index of food-anticipatory activity (FAA). Both groups entrained to the daily meal with an increase in feeder approach time several hours before meal onset. There were no significant differences in the timing or the amount of FAA between groups. Furthermore, FAA was maintained during 3 days of food deprivation in both groups. In accordance with previous studies, the results show that olfactory cues are not necessary for the entrainment of FAA.

Extraordinary behavioral flexibility is necessary in order to meet energy needs. An animal must know when and where to search for food, guard itself and its offspring against predators, and continuously adjust foraging behavior as the changing environment dictates. These pressures have selected for the ability to accomplish numerous functions simultaneously. This is particularly necessary during the execution of motivated behaviors (behaviors that have initiation, procurement, and consummatory phases) (Swanson 2000), and is facilitated by habitual performance. Specifically, while motivated learning initially requires significant cognitive effort, with practice, complex tasks can be performed quite automatically. In a recent study, Hernandez et al. (2006) examined the neurobiology of this transition, from early learning to automatic responding, based on the hypothesis that the transition is mediated by plasticity within neural circuitry regulating motivated learning. Hernandez et al. assessed neural plasticity anatomically using in situ hybridization to measure changes in the expression of two immediate early genes important for learning and memory: Zif268 and Homer 1a. Zif268 is a transcription factor involved in cellular and behavioral assays of learning (Knapska and Kaczmarek 2004). Homer 1a is a component of the postsynaptic density of glutamate synapses (de Bartolomeis and Iasevoli 2003). This is especially important given the role played by glutamate transmission in learning (Gubellini et al. 2004; Kelley 2004). In addition to a general involvement in learning and memory, Zif268 and Homer 1a are also specifically important for motivated learning (Thomas et al. 2003; Lominac et al. 2005; Petrovich et al. 2005; Szumlinski et al. 2006). These genes were therefore chosen to examine whether plasticity within motivational brain circuitry is associated with the transition from initial motivated responding to automatic responding. Rats learned an operant task where lever presses produced sugar pellets. In one group, gene expression was measured after several days of training, in which behavior was reliable but responding had not reached its maximum. These subjects were referred to as “early learners.” In a separate group, gene expression was measured several weeks later, well after the behavioral measure had reached its maximum level, and these subjects were referred to as “overtrained.” Control subjects were exposed to the same conditions; however, lever pressing had no programmed consequences. Relative to controls, early learners showed significantly higher expression levels of Zif268 and Homer 1a in several frontal cortical regions (including anterior cingulate, orbitofrontal, and prefrontal). In contrast with the early learners, overtrained subjects had significantly lower Homer 1a expression in these cortical regions compared with controls (there was no difference for Zif268). This reversal was only found in the frontal cortex and not in other areas where gene expression was elevated in early learners, such as the dorsomedial striatum, hippocampus, or sensory and motor cortex. These findings suggest decreased cortical influence during automatic responding. Hernandez et al. (2006) then directly compared levels of gene expression in overtrained subjects to early learners. Overtrained rats had significantly higher Homer 1a expression in the ventrolateral striatum compared to early learners. Further, there was a positive correlation between the number of lever presses and Homer 1a expression in the ventrolateral striatum. These findings suggest that this specific portion of the striatum plays an important role in automation of behavior. And together, data presented in Hernandez et al. suggest a “dynamic shift” in the expression of plasticity-related immediate early gene expression, in which there is decreased cortical and increased striatal activity as a specific motivated output becomes very well established. Importantly, this “cortex to striatum” shift may be unique to motivational learning, as a fundamentally distinct mechanism has been identified for spatial and contextual memory storage. Several recent studies have used similar neuroanatomical methods, as used by Hernandez et al. (2006) to convincingly demonstrate that recent spatial memories involve processing within the hippocampus, whereas remote spatial memories (∼30 d after learning) require cortical processing (Frankland et al. 2004; Maviel et al. 2004; Frankland and Bontempi 2005). These studies demonstrate differential neural regulation of learning and memory depending on the behavior being examined. Dynamic neuroplasticity mediating transitions from early learning to automatic responding is important for understanding the evolution of motivated behavior. Approach and avoidance behaviors (Adler 1966; Zhang et al. 2005), as well as operant learning (Brembs 2003; Hawkins et al. 2006), are ancient processes predating neocortex. For food seeking in mammals, the ability to distinguish between positive and negative valence is processed in phylogenetically older brain regions in the midand hindbrain (Grill and Norgren 1978), areas expressing very low levels of plasticity-related genes (Kelley 2004). However, these regions are highly interconnected to the more phylogenetically recent structures, such as neocortex and subcortical forebrain structures like the striatum. These regions possess the neuroplastic capabilities to efficiently meet energy needs by orchestrating the appropriate influence over hard-wired feeding areas, depending on level of experience of a particular motivated output. Specifically, with practice a complex behavior can be executed with only a minimal burden of awareness, because it is under striatal regulation. This leaves the cortex free to process additional features of the foraging experience, such as sensing danger or changes in the environment that may signal more promising foraging opportunities. If true, such shifts in corticostriatal regulation of motivated behavior can explain the adaptive benefit of habitual behavior. Perhaps the most significant aspect of highly plastic capabilities of the coritcostriatal system mediating motivated learning is that this system is extremely sensitive to drugs of abuse (Nestler 2004). Drug addiction is widely conceptualized as a disorder of learning and memory (Di Chiara 1999; Hyman 2005), and increasing evidence suggests that compulsive drug seeking is 1Corresponding author. E-mail rcarelli@unc.edu; fax (919) 962-2537. Article is online at http://www.learnmem.org/cgi/doi/10.1101/lm.398806. Commentary