Prelimbic Cortex Activity during a Distress Tolerance Task Predicts Cocaine-Seeking Behavior in Male, But Not Female Rats.

Distress tolerance (DT) is defined as the ability to persist in challenging goal-directed behavior in the face of stress, and individuals with low DT exhibit heightened drug-seeking behavior. However, no preclinical studies have examined the neurobiology underlying this phenomenon. To assess this, in vivo electrophysiology was used in Long Evans male and female rats during a DT task to record neural activity in the prelimbic cortex (PrL), a brain region implicated in drug-seeking. Rats were first assessed for DT, defined as the amount of time elapsed before rats quit seeking reward in an increasingly difficult operant task. Subsequently, rats underwent 2 weeks of self-administration for either water/saline or cocaine for 6 h/day. Animals then began a 1 month period of experimenter-imposed abstinence to induce heightened drug-seeking behavior. On day 28 of abstinence, DT and neural activity were reassessed; and on day 30, cocaine-seeking behavior was examined under extinction. Males had significantly higher DT than females and exhibited significantly more phasic PrL activity during the DT task. Furthermore, in male rats with a history of cocaine, PrL activity shifted to track DT; and this change in activity significantly correlated with the change in DT. Additionally, male (but not female) rats with low DT after 28 d of abstinence had significantly heightened drug-seeking behavior. Finally, PrL activity during the DT task predicted cocaine-seeking behavior. Collectively, these data demonstrate an important role for the PrL in DT in males, and link this neural activity and behavior to drug-seeking, particularly in males.SIGNIFICANCE STATEMENT Distress tolerance (DT) is defined as the ability to persist in challenging goal-directed behavior in the face of stress, and individuals with low DT exhibit heightened drug-seeking. Here, we investigated the role of the prelimbic cortex (PrL) in DT and its relationship to cocaine-seeking in male and female rats. We found that males had significantly higher DT than females and exhibited significantly more PrL activity during the DT task. Furthermore, male (but not female) rats with low DT after 28 d of abstinence had significantly heightened drug-seeking behavior. Finally, PrL activity during the DT task predicted cocaine-seeking. These data demonstrate an important role for the PrL in DT and link this neural activity and behavior to drug-seeking in males.

Prelimbic Cortical Neurons Track Preferred Reward Value and Reflect Impulsive Choice during Delay Discounting Behavior.

In delay discounting, individuals discount the value of a reward based on the delay to its receipt. The prelimbic cortex (PrL) is heavily interconnected with several brain regions implicated in delay discounting, but the specific contributions of the PrL to delay discounting are unknown. Here, we used multineuron electrophysiological recording methods in Long-Evans male (n = 10) and female (n = 9) rats to characterize the firing dynamics of PrL neurons during discrete cue and lever press events in a delay discounting task. Rats' initial preference for the large reward decreased as delays for that outcome increased across blocks, reflecting classic discounting behavior. Electrophysiological recordings revealed that subgroups of neurons exhibited phasic responses to cue presentations and lever presses. These phasic neurons were found to respond to either large/delay, small/immediate, or both trial types and the percentage of these neurons shifted across blocks as the expected value of the reward changed. Critically, this shift was only seen during trials in which animals could choose their preferred option (free choice trials) and not during trials where animals could choose only one option (forced choice trials). Further, this shift was dependent on rats' inherent impulsivity because high impulsive rats demonstrated a greater percentage of small/immediate-responsive neurons as the task progressed. Collectively, these findings suggest a unique role for the PrL in encoding reward value during delay discounting that is influenced by individual differences in impulsivity.SIGNIFICANCE STATEMENT In delay discounting, individuals discount the value of a reward based on the delay to its receipt. Here, we used electrophysiology to investigate the role of the prelimbic cortex (PrL) in this process. We found that subsets of neurons shifted activity as a function of the changing expected delay and reward magnitude, but this shift was only evident during trials in which animals could choose their preferred option. Further, this dynamic neural activity depended on rats' inherent impulsivity, with impulsive rats exhibiting a stronger neural shift toward the immediate reward as the task progressed. These findings suggest a role for the PrL in encoding reward value during delay discounting that is influenced by goal-directed context and individual differences in impulsivity.

Nucleus accumbens shell dopamine mediates outcome value, but not predicted value, in a magnitude decision-making task.

Effective decision-making depends on an animal's ability to predict and select the outcome of greatest value, and the nucleus accumbens (NAc) and its dopaminergic input play a key role in this process. We previously reported that rapid dopamine release in the NAc shell preferentially tracks the "preferred" (i.e., large reward) option during cues that predict the ability to respond for rewards of different sizes, as well as during reward delivery itself. The present study assessed whether shell dopamine release at these discrete times selectively mediated choice behavior for rewards of different magnitudes using optogenetics. Here, using Long Evans TH:Cre± rats we employed selective optogenetic stimulation of dopamine terminals in the NAc shell during either reward-predictive cues (experiment 1) or reward delivery (experiment 2) in a magnitude-based decision-making task. We found that in TH:Cre± rats, but not littermate controls, optical stimulation during low-magnitude reward delivery during forced choice trials was sufficient to bias preference for this option when given a choice. In contrast, optical stimulation of shell dopamine terminals during low-magnitude reward-predictive cues in forced choice trials did not shift free choice behavior in TH:Cre± rats or controls. The findings indicate that preferential dopamine signaling in the NAc shell during reward outcome (delivery), but not reward-predictive cues are sufficient to influence choice behavior in our task supporting a causal role of dopamine in the NAc shell in reward outcome value, but not value-based predictive strategies.

A Neuronal Ensemble in the Rostral Agranular Insula Tracks Cocaine-Induced Devaluation of Natural Reward and Predicts Cocaine Seeking.

In substance use disorders, negative affect associated with drug withdrawal can elicit strong drug craving and promote relapse. One brain region implicated in those processes is the rostral agranular insular cortex (RAIC), although precisely how this region encodes negative affect associated with drug seeking is unknown. Here, a preclinical model was used where RAIC activity was examined in male Sprague Dawley rats during intraoral infusions of a sweet (saccharin) paired with impending but delayed access to cocaine self-administration, and for comparative purposes, during the sweet predicting saline self-administration or injection of lithium chloride (LiCl), or during intraoral infusions of a bitter taste (quinine). Consistent with previous work, cocaine-paired saccharin, LiCl-paired saccharin, and quinine all elicited aversive taste reactivity. However, the aversive taste reactivity elicited by the cocaine-paired tastant was qualitatively different from that evoked by the other two agents. Furthermore, differences in taste reactivity were reflected in RAIC cell firing, where distinct shifts in neural signaling were observed specifically after cocaine but not LiCl conditioning. Notably, low motivation for cocaine (indicated by low loading and slower latencies to lever press) was correlated with this shift in RAIC signaling, but aversive (gaping) responses were not. Collectively, these findings indicate that cocaine-paired tastants elicit unique aspects of aversive behaviors that differ from traditional conditioned taste aversion (LiCl) or quinine and that the RAIC plays a role in modulating drug-seeking behaviors driven by drug-induced dysphoria (craving), but not negative affect per se.SIGNIFICANCE STATEMENT In substance use disorders, negative affect associated with drug cues can elicit craving and promote relapse; however, the underlying neurocircuitry of this phenomenon is unknown. Here, we investigated the role of the rostral agranular insula cortex (RAIC) in these processes using a preclinical model wherein intraoral delivery of a sweet is paired with delayed access to cocaine self-administration. The taste comes to elicit negative affect that predicts heightened drug seeking. Here, we found that a population of RAIC neurons became inhibited during presentation of the cocaine-paired tastant (when negative affect is high) and that this inhibitory neural profile predicted lower drug seeking. These findings suggest that the RAIC may function to oppose cue-induced cocaine craving and help reduce motivation for the drug.

Low distress tolerance predicts heightened drug seeking and taking after extended abstinence from cocaine self-administration.

Distress tolerance (DT), defined as the ability to persist in goal-directed behavior while experiencing psychological distress, is associated with greater frequency of substance use and poor treatment outcomes. To examine a potential causal role substance use may play in DT, we developed a rodent model of DT in which rats had to press a lever within a continuously decreasing time window for reward while receiving negative feedback on failure trials. DT was defined as the time rats continued to seek reward before quitting the task. We assessed the relationship of DT with cocaine seeking/taking by measuring DT before cocaine self-administration (SA), and after 1 week and 1 month of drug abstinence. We found that DT prior to cocaine SA did not predict cocaine seeking/taking, yet DT measured after 1 month abstinence significantly predicted subsequent high levels of early session cocaine taking. Additionally, high DT measured after abstinence protected against high cocaine seeking, but this protective effect was blocked in rats with high impulsivity. Finally, while a decrease in 1 month-abstinent DT was observed following SA across treatment conditions, among cocaine-exposed rats, greater cocaine SA correlated with a steeper decrease in DT. These results show that low DT after drug abstinence is associated with heightened levels of cocaine seeking and taking behavior and that impulsivity influences this effect. Collectively, these results support the validity of our rodent DT model while extending the human literature and set the foundation for future animal studies designed to determine neural mechanisms underlying DT.

Mice bred for severity of acute alcohol withdrawal respond differently in a go/no-go task.

BACKGROUND: Mice selectively bred for high or low withdrawal to acute alcohol differ on a number of traits, including consumption of alcohol, conditioned place preference for alcohol, and sensitivity to alcohol-induced locomotor activity. One trait that has not been examined in these mice is behavioral inhibition. METHODS: High and low alcohol withdrawal mice (second replicate: high and low acute alcohol withdrawal [HAW-2/LAW-2]) were trained and tested in a Go/No-go task. Mice were administered 0.0, 0.5, 1.0, and 1.5 g/kg ethanol (EtOH) on 3 occasions according to an incomplete Latin Square. A separate cohort of C57BL/6J (B6) and DBA/2J (D2) mice (the progenitor strains for HAW-2/LAW-2 mice) underwent the same protocol, using the same EtOH doses. RESULTS: HAW-2 and LAW-2 mice did not differ in behavioral inhibition at baseline, although LAW-2 mice did have higher overall levels of responding in the task. EtOH did not alter behavioral inhibition in these mice. However, it did decrease responses to the Go cue, and this effect was greater in HAW-2 mice than in LAW-2 mice. D2 mice had lower behavioral inhibition than B6 mice at baseline, and EtOH slightly decreased behavioral inhibition in both strains. CONCLUSIONS: The findings with D2 and B6 mice generally fit with the existing literature. However, the lack of a difference in behavioral inhibition between HAW-2 and LAW-2 mice was unexpected, as well as the absence of any effect of these doses of EtOH on behavioral inhibition in these mice. Nonetheless, the findings do suggest that selectively breeding for high or low withdrawal to acute alcohol can lead to differences in operant behavior in the Go/No-go task.

Ovarian hormones and the heterogeneous receptor mechanisms mediating the discriminative stimulus effects of ethanol in female rats.

Past studies have suggested that progesterone-derived ovarian hormones contribute to the discriminative stimulus effects of ethanol, particularly via progesterone metabolites that act at γ-aminobutyric acid type A (GABA(A)) receptors. It is unknown whether loss of ovarian hormones in women, for example, after menopause, may be associated with altered receptor mediation of the effects of ethanol. The current study measured the substitution of allopregnanolone, pregnanolone, pentobarbital, midazolam, dizocilpine, TFMPP, and RU 24969 in female sham and ovariectomized rats trained to discriminate 1.0 g/kg ethanol from water. The groups did not differ in the substitution of GABA(A)-positive modulators (barbiturates, benzodiazepines, neuroactive steroids) or the N-methyl-D-aspartate receptor antagonist dizocilpine. Similarly, blood-ethanol concentration did not differ between the groups, and plasma adrenocorticotropic hormone, progesterone, pregnenolone, and deoxycorticosterone were unchanged 30 min after administration of 1.0 g/kg ethanol or water. However, substitution of neuroactive steroids and RU 24969, a 5-hydroxytryptamine (5-HT)(1A/1B) receptor agonist, was lower than observed in previous studies of male rats, and TFMPP substitution was decreased in ovariectomized rats. Ovarian hormones appear to contribute to 5-HT receptor mediation of the discriminative stimulus effects of ethanol in rats.

Acute ethanol administration and reinforcer magnitude reduction both reduce responding and increase response latency in a go/no-go task.

BACKGROUND: Ethanol (EtOH) administration decreases behavioral inhibition in human subjects, assessed using cued Go/No-Go tasks, in which an unreliable cue suggests whether participants will be required to respond or not when a signal occurs. Few studies have examined EtOH's effects on behavioral inhibition in animals, and those that have done so have used Go/No-Go tasks in which no warning cue was provided. METHODS: Two cohorts of male Long-Evans rats were trained and tested on 2 different Go/No-Go procedures with differing ratios of Go to No-Go trials (25 to 75 and 50 to 50). Using a within-subjects design, each rat was administered 0.0, 0.63, 0.95, and 1.27 g/kg of EtOH (i.p.) on 3 separate occasions according to an incomplete Latin square. An additional experiment examined the effects of reducing the amount of sucrose given for correct responses to either the Go or the No-Go signal in the absence of EtOH administration. RESULTS: Acute intraperitoneal EtOH administration dose-dependently decreased responding during the No-Go signal (false alarms), the Go signal (hits), and responding prior to the occurrence of either signal (precue response rate). These effects were more pronounced in rats with the 50 to 50 ratio. Reducing the amount of sucrose presented generally led to a decrease in responding, although this effect was also moderated by the Go to No-Go ratio employed and the contingency relationship (reduced sucrose for correct Go trial responding or for correct No-Go trial response withholding). CONCLUSIONS: Acute EtOH administration does not decrease behavioral inhibition in rats in this task. Rather EtOH appears to dose-dependently decrease behavior in general, possibly by reducing the efficacy of the sucrose reinforcer, as both EtOH administration and sucrose reduction for Go trials yielded similar patterns of behavioral responding in this task in rats.

An opposing role for prelimbic cortical projections to the nucleus accumbens core in incubation of craving for cocaine versus water.

BACKGROUND: Both drug and natural reward-seeking have been shown to increase following an extended period of abstinence, a phenomenon termed 'incubation of craving'. Although this phenomenon involves many brain regions, the projections from the prelimbic cortex (PrL) to the nucleus accumbens (NAc) core have been strongly implicated in incubation of cocaine-seeking. However, this circuit has not been investigated in the context of incubation of craving for natural rewards. METHODS: Male Long Evans rats were trained to self-administer cocaine or water/saline 6 h/d for 14 days and subsequently entered 1 month of experimenter-imposed abstinence. Rats then underwent an optogenetic stimulation protocol used to induce long term depression in the PrL terminals to the NAc core immediately before beginning an extinction test used to assess incubation of craving. RESULTS: Control cocaine rats showed heightened drug-seeking on day 30 when compared to day 1 of abstinence, demonstrating incubation of craving. Notably, optogenetic stimulation of the PrL to NAc core pathway blocked this behavior in cocaine rats. In contrast, optogenetic stimulation of the PrL to NAc core pathway induced incubation of craving in water/saline rats. CONCLUSIONS: These findings suggest that neuroadaptations in the PrL to NAc core pathway play opposing roles in the incubation of craving for cocaine versus water.

A sex-dependent role for the prelimbic cortex in impulsive action both before and following early cocaine abstinence.

Although impulsive action is strongly associated with addiction, the neural underpinnings of this relationship and how they are influenced by sex have not been well characterized. Here, we used a titrating reaction time task to assess differences in impulsive action in male and female Long Evans rats both before and after short (4-6 days) or long (25-27 days) abstinence from 2 weeks of cocaine or water/saline self-administration (6 h daily access). Neural activity in the prelimbic cortex (PrL) and nucleus accumbens (NAc) core was assessed at each time point. We found that a history of cocaine self-administration increased impulsivity in all rats following short, but not long, abstinence. Furthermore, male rats with an increased ratio of excited to inhibited neurons in the PrL at the start of each trial in the task exhibited higher impulsivity in the naïve state (before self-administration). Following short abstinence from cocaine, PrL activity in males became more inhibited, and this change in activity predicted the shift in impulsivity. However, PrL activity did not track impulsivity in female rats. Additionally, although the NAc core tracked several aspects of behavior in the task, it did not track impulsivity in either sex. Together, these findings demonstrate a sex-dependent role for the PrL in impulsivity both before and after a history of cocaine.

Selective nicotinic acetylcholine receptor agonists: potential therapies for neuropsychiatric disorders with cognitive dysfunction.

Cognitive impairment is one of the most functionally debilitating aspects of neuropsychiatric and neurodegenerative disorders, such as schizophrenia and Alzheimer's disease, despite treatment with available pharmacotherapies. There is emerging evidence that nicotine improves cognitive function in humans and nonhuman species and this has sparked interest in the development of novel nicotinic treatments for cognitive dysfunction. The examination of selective alpha7 and alpha4beta2 nicotinic acetylcholine receptor (nAChR) agonists suggests that both receptor subtypes mediate improvement in attention, learning and working memory. When compared with available pharmacotherapies, specific nAChR agonists may represent unique targets for the treatment of neuropsychiatric and neurodegenerative disorders that feature cognitive impairment as a key symptom.

Impulsive Rats Exhibit Blunted Dopamine Release Dynamics during a Delay Discounting Task Independent of Cocaine History.

The inability to wait for a large, delayed reward when faced with a small, immediate one, known as delay discounting, has been implicated in a number of disorders including substance abuse. Individual differences in impulsivity on the delay discounting task are reflected in differences in neural function, including in the nucleus accumbens (NAc) core. We examined the role of a history of cocaine self-administration, as well as individual differences in impulsivity, on rapid dopamine (DA) release dynamics in the NAc core. Rats with a history of cocaine or water/saline self-administration were tested on delay discounting while being simultaneously assayed for rapid DA release using electrochemical methods. In controls, we found that cue DA release was modulated by reward delay and magnitude, consistent with prior reports. A history of cocaine had no effect on either delay discounting or DA release dynamics. Nonetheless, independent of drug history, individual differences in impulsivity were related to DA release in the NAc core. First, high impulsive animals exhibited dampened cue DA release during the delay discounting task. Second, reward delay and magnitude in high impulsive animals failed to robustly modulate changes in cue DA release. Importantly, these two DAergic mechanisms were uncorrelated with each other and, together, accounted for a high degree of variance in impulsive behavior. Collectively, these findings demonstrate two distinct mechanisms by which rapid DA signaling may influence impulsivity, and illustrate the importance of NAc core DA release dynamics in impulsive behavior.

Going for broke: dopamine influences risky choice.

Dopamine neurons track reward by increasing or decreasing their firing rate when a reward is present or absent. In this issue of Neuron, Stopper et al. (2014) demonstrate that artificially eliminating these dopamine bursts or dips can alter risky decision-making.

Partial inactivation of nucleus accumbens core decreases delay discounting in rats without affecting sensitivity to delay or magnitude.

Increased preference for smaller, sooner rewards (delay discounting) is associated with several behavioral disorders, including ADHD and substance use disorders. However, delay discounting is a complex cognitive process and the relationship is unclear between the pathophysiology of the disorders and the component processes underlying delay discounting, including sensitivity to reinforcer delay and sensitivity to reinforcer magnitude. To investigate these processes, male Long Evans rats were trained in one of three tasks measuring sensitivity to delay, sensitivity to magnitude, or both (typical delay discounting task). After learning the task, animals were implanted with bilateral cannulae into either the nucleus accumbens core (AcbC) or the lateral orbitofrontal cortex (lOFC), both of which have been implicated in delay discounting. Upon recovering from the surgery, a baclofen/muscimol cocktail was infused to temporarily inactivate each of these two regions and task performance was assessed. Unlike previous studies showing that lesions of the AcbC increased delay discounting, partial inactivation of the AcbC decreased delay discounting, although it had no effects on the tasks independently assessing either sensitivity to delay or magnitude. The effects of AcbC inactivation were larger in animals that had low levels of delay discounting at baseline. Inactivation of the lOFC had no effects on behavior in any task. These findings suggest that the AcbC may act to promote impulsive choice in individuals with low impulsivity. Furthermore, the data suggest that the AcbC is able to modulate delay and magnitude sensitivity together, but not either of the two in isolation.

Behavioral disinhibition in mice bred for high drinking in the dark (HDID) and HS controls increases following ethanol.

BACKGROUND: Alcohol consumption and behavioral inhibition share some common underlying genetic mechanisms. The current study examined whether lines of mice selected for high blood ethanol concentrations, attained by heavy drinking in the dark period (DID) of the light-dark cycle that models binge drinking, also exhibit higher levels of drug-naïve inhibition. It also examined whether the administration of ethanol would result in higher levels of disinhibition in these selected lines compared to the founder stock (HS). METHODS: A Go/No-Go task was used to assess baseline inhibition and the effects of acute ethanol on disinhibition (response to a No-Go cue) in the HS line and in mice selected for high levels of DID (HDID-1 and HDID-2). RESULTS: Lines did not differ in inhibition at baseline and all lines showed increased disinhibition following moderate doses of ethanol. Ethanol decreased responding to Go cues for HDID-2 and HS lines at high doses but not HDID-1 mice. CONCLUSIONS: These data corroborate previous work showing ethanol-induced increases in behavioral disinhibition. The selection paradigm did not result in differential sensitivity to the disinhibiting effects of ethanol, but did result in differential sensitivity to the suppressant effects of ethanol on operant behavior between the two HDID lines.

Sensitivity to reinforcer delay predicts ethanol's suppressant effects, but itself is unaffected by ethanol.

BACKGROUND: Relative preference for smaller, sooner rewards over larger, later rewards ("delay discounting") is increased by acute ethanol. Additionally, drug-naïve levels of delay discounting can predict subsequent ethanol consumption. However, it is unknown whether these phenomena are driven by a difference in sensitivity to the reinforcer delay or a difference in sensitivity to the reinforcer magnitude, because typical delay discounting tasks manipulate both parameters simultaneously. METHODS: To disambiguate these factors, two tasks were developed in which animals chose between levers with either different delay contingencies (adjusting delay task) or different magnitude contingencies (adjusting magnitude task). When task performance was stable, rats received ethanol (0, 0.6, and 0.9 g/kg, i.p.). RESULTS: Ethanol did not affect sensitivity to delay or sensitivity to magnitude. However, responding was suppressed at the highest dose of ethanol (0.9 g/kg). Less suppression was found in animals exhibiting high levels of drug-naïve sensitivity to delay. CONCLUSION: Thus, this study suggests that ethanol's effect on standard delay discounting tasks is not due to an alteration in sensitivity to delay or magnitude. Additionally, these data show that animals with high sensitivity to delay are resistant to the behaviorally suppressant effects of ethanol, which suggests that low tolerance for delayed rewards and low sensitivity to the behaviorally suppressant effects of ethanol may partly be driven by the same underlying mechanism.

Behavioral inhibition in mice bred for high vs. low levels of methamphetamine consumption or sensitization.

RATIONALE: Research indicates that genetics influence methamphetamine self-administration as well as sensitization to the psychomotor-stimulating effects of methamphetamine (MA). Other studies have suggested that heightened levels of impulsivity, including low levels of behavioral inhibition, are associated with the use of drugs, including MA. OBJECTIVES: The current study examined whether lines of mice selected for traits associated with a heightened risk of developing MA dependence would also exhibit low levels of drug-naïve inhibition and whether administration of MA would result in different levels of inhibition in animals selected to consume or respond more to MA. METHODS: A go/no-go task was used to assess inhibition in male and female mice selected for low or high levels of MA consumption or selected for high or low levels of locomotor sensitization to repeated injections of MA. RESULTS: Mice selected for MA sensitization differed in false alarms, precue response rates (measures of behavioral inhibition), and also hits (measure of operant responding). Mice selected for MA consumption did not differ in measures of behavioral inhibition, though hits differed. When MA was administered prior to the task, false alarms, precue response rates, and hits decreased for mice from all selected lines. Female high drinking mice were particularly resistant to MA's effects on hits, but not precue response rate or false alarms. CONCLUSIONS: These data suggest a shared, but complex, genetic association between inhibition processes, general levels of operant responding, and MA sensitization or consumption.