How changes in dopamine D2 receptor levels alter striatal circuit function and motivation.

It was first posited, more than five decades ago, that the etiology of schizophrenia involves overstimulation of dopamine receptors. Since then, advanced clinical research methods, including brain imaging, have refined our understanding of the relationship between striatal dopamine and clinical phenotypes as well as disease trajectory. These studies point to striatal dopamine D2 receptors, the main target for all current antipsychotic medications, as being involved in both positive and negative symptoms. Simultaneously, animal models have been central to investigating causal relationships between striatal dopamine D2 receptors and behavioral phenotypes relevant to schizophrenia. We begin this article by reviewing the circuit, cell-type and subcellular locations of dopamine D2 receptors and their downstream signaling pathways. We then summarize results from several mouse models in which D2 receptor levels were altered in various brain regions, cell-types and developmental periods. Behavioral, electrophysiological and anatomical consequences of these D2 receptor perturbations are reviewed with a selective focus on striatal circuit function and alterations in motivated behavior, a core negative symptom of schizophrenia. These studies show that D2 receptors serve distinct physiological roles in different cell types and at different developmental time points, regulating motivated behaviors in sometimes opposing ways. We conclude by considering the clinical implications of this complex regulation of striatal circuit function by D2 receptors.

Dopamine D2 receptors modulate the cholinergic pause and inhibitory learning.

Cholinergic interneurons (CINs) in the striatum respond to salient stimuli with a multiphasic response, including a pause, in neuronal activity. Slice-physiology experiments have shown the importance of dopamine D2 receptors (D2Rs) in regulating CIN pausing, yet the behavioral significance of the CIN pause and its regulation by dopamine in vivo is still unclear. Here, we show that D2R upregulation in CINs of the nucleus accumbens (NAc) lengthens the pause in CIN activity ex vivo and enlarges a stimulus-evoked decrease in acetylcholine (ACh) levels during behavior. This enhanced dip in ACh levels is associated with a selective deficit in the learning to inhibit responding in a Go/No-Go task. Our data demonstrate, therefore, the importance of CIN D2Rs in modulating the CIN response induced by salient stimuli and point to a role of this response in inhibitory learning. This work has important implications for brain disorders with altered striatal dopamine and ACh function, including schizophrenia and attention-deficit hyperactivity disorder (ADHD).

Developmental impact of glutamate transporter overexpression on dopaminergic neuron activity and stereotypic behavior.

Obsessive-compulsive disorder (OCD) is a disabling condition that often begins in childhood. Genetic studies in OCD have pointed to SLC1A1, which encodes the neuronal glutamate transporter EAAT3, with evidence suggesting that increased expression contributes to risk. In mice, midbrain Slc1a1 expression supports repetitive behavior in response to dopaminergic agonists, aligning with neuroimaging and pharmacologic challenge studies that have implicated the dopaminergic system in OCD. These findings suggest that Slc1a1 may contribute to compulsive behavior through altered dopaminergic transmission; however, this theory has not been mechanistically tested. To examine the developmental impact of Slc1a1 overexpression on compulsive-like behaviors, we, therefore, generated a novel mouse model to perform targeted, reversible overexpression of Slc1a1 in dopaminergic neurons. Mice with life-long overexpression of Slc1a1 showed a significant increase in amphetamine (AMPH)-induced stereotypy and hyperlocomotion. Single-unit recordings demonstrated that Slc1a1 overexpression was associated with increased firing of dopaminergic neurons. Furthermore, dLight1.1 fiber photometry showed that these behavioral abnormalities were associated with increased dorsal striatum dopamine release. In contrast, no impact of overexpression was observed on anxiety-like behaviors or SKF-38393-induced grooming. Importantly, overexpression solely in adulthood failed to recapitulate these behavioral phenotypes, suggesting that overexpression during development is necessary to generate AMPH-induced phenotypes. However, doxycycline-induced reversal of Slc1a1/EAAT3 overexpression in adulthood normalized both the increased dopaminergic firing and AMPH-induced responses. These data indicate that the pathologic effects of Slc1a1/EAAT3 overexpression on dopaminergic neurotransmission and AMPH-induced stereotyped behavior are developmentally mediated, and support normalization of EAAT3 activity as a potential treatment target for basal ganglia-mediated repetitive behaviors.

Prior experience modifies acquisition trajectories via response-strategy sampling.

Few studies have considered how signal detection parameters evolve during acquisition periods. We addressed this gap by training mice with differential prior experience in a conditional discrimination, auditory signal detection task. Naïve mice, mice given separate experience with each of the later correct choice options (Correct Choice Response Transfer, CCRT), and mice experienced in conditional discriminations (Conditional Discrimination Transfer, CDT) were trained to detect the presence or absence of a tone in white noise. We analyzed data assuming a two-period model of acquisition: a pre-solution and solution period (Heinemann EG (1983) in The Presolution period and the detection of statistical associations. In: Quantitative analyses of behavior: discrimination processes, vol. 4, pp. 21-36). Ballinger. http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.536.1978andrep=rep1andtype=pdf ). The pre-solution period was characterized by a selective sampling of biased response strategies until adoption of a conditional responding strategy in the solution period. Correspondingly, discriminability remained low until the solution period; criterion took excursions reflecting response-strategy sampling. Prior experience affected the length and composition of the pre-solution period. Whereas CCRT and CDT mice had shorter pre-solution periods than naïve mice, CDT and Naïve mice developed substantial criterion biases and acquired asymptotic discriminability faster than CCRT mice. To explain these data, we propose a learning model in which mice selectively sample and test different response-strategies and corresponding task structures until they exit the pre-solution period. Upon exit, mice adopt the conditional responding strategy and task structure, with action values updated via inference and generalization from the other task structures. Simulations of representative mouse data illustrate the viability of this model.

Overexpression of striatal D2 receptors reduces motivation thereby decreasing food anticipatory activity.

Dopamine has been implicated in circadian timing underlying the food entrainable oscillator (FEO) circuitry and overexpression of the dopamine D2 receptor (D2R) in the striatum has been reported to reduce motivation to obtain food rewards in operant tasks. In the present study, we explored both of these mechanisms by examining food anticipatory activity (FAA) in dopamine D2 receptor-overexpressing (D2R-OE) mice under various durations of food availability. First, we noted that at baseline, there were no differences between D2R-OE mice and their littermates in activity level, food intake, and body weight or in circadian activity. Under conditions of very restricted food availability (4 or 6 hr), both genotypes displayed FAA. In contrast, under 8-hr food availability, control mice showed FAA, but D2R-OE mice did not. Normalization of D2R by administration of doxycycline, a tetracycline analogue, rescued FAA under 8-hr restricted food. We next tested for circadian regulation of FAA. When given ad libitum access to food, neither D2R-OE nor controls were active during the daytime. However, after an interval of food restriction, all mice showed elevated locomotor activity at the time of previous food availability in the day, indicating circadian timing of anticipatory activity. In summary, motivation is reduced in D2R-OE mice but circadian timing behavior is not affected. We conclude that an increase in striatal D2R reduces FAA by modulating motivation and not by acting on a clock mechanism.

An Interaction between Serotonin Receptor Signaling and Dopamine Enhances Goal-Directed Vigor and Persistence in Mice.

The functionally selective 5-HT2C receptor ligand SB242084 can increase motivation and have rapid onset anti-depressant-like effects. We sought to identify the specific behavioral effects of SB242084 treatment and elucidate the mechanism in female and male mice. Using a quantitative behavioral approach, we determined that SB242084 increases the vigor and persistence of goal-directed activity across different types of physical work, particularly when work requirements are demanding. We found this influence of SB242084 on effort, rather than reward to be reflected in striatal DA measured during behavior. Using in vivo fast scan cyclic voltammetry, we found that SB242084 has no effect on reward-related phasic DA release in the NAc. Using in vivo microdialysis to measure tonic changes in extracellular DA, we also found no changes in the NAc. In contrast, SB242084 treatment increases extracellular DA in the dorsomedial striatum, an area that plays a key role in response vigor. These findings have several implications. At the behavioral level, this work shows that the capacity to work in demanding situations can be increased, without a generalized increase in motor activity or reward value. At the circuit level, we identified a pathway restricted potentiation of DA release and showed that this was the reason for the increased response vigor. At the cellular level, we show that a specific serotonin receptor cross talks to the DA system. Together, this information provides promise for the development of treatments for apathy, a serious clinical condition that can afflict patients with psychiatric and neurological disorders.SIGNIFICANCE STATEMENT Motivated behaviors are modulated by reward value, effort demands, and cost-benefit computations. This information drives the decision to act, which action to select, and the intensity with which the selected action is performed. Because these behavioral processes are all regulated by DA signaling, it is very difficult to influence selected aspects of motivated behavior without affecting others. Here we identify a pharmacological treatment that increases the vigor and persistence of responding in mice, without increasing generalized activity or changing reactions to rewards. We show that the 5-HT2C-selective ligand boosts motivation by potentiating activity-dependent DA release in the dorsomedial striatum. These results reveal a novel strategy for treating patients with motivational deficits, avolition, or apathy.

Decreasing Striatopallidal Pathway Function Enhances Motivation by Energizing the Initiation of Goal-Directed Action.

UNLABELLED: Altered dopamine D2 receptor (D2R) binding in the striatum has been associated with abnormal motivation in neuropsychiatric disorders, including schizophrenia. Here, we tested whether motivational deficits observed in mice with upregulated D2Rs (D2R-OEdev mice) are reversed by decreasing function of the striatopallidal "no-go" pathway. To this end, we expressed the Gαi-coupled designer receptor hM4D in adult striatopallidal neurons and activated the receptor with clozapine-N-oxide (CNO). Using a head-mounted miniature microscope we confirmed with calcium imaging in awake mice that hM4D activation by CNO inhibits striatopallidal function measured as disinhibited downstream activity in the globus pallidus. Mice were then tested in three operant tasks that address motivated behavior, the progressive ratio task, the progressive hold-down task, and outcome devaluation. Decreasing striatopallidal function in the dorsomedial striatum or nucleus accumbens core enhanced motivation in D2R-OEdev mice and control littermates. This effect was due to increased response initiation but came at the cost of goal-directed efficiency. Moreover, response vigor and the sensitivity to changes in reward value were not altered. Chronic activation of hM4D by administering CNO for 2 weeks in drinking water did not affect motivation due to a tolerance effect. However, the acute effect of CNO on motivation was reinstated after discontinuing chronic treatment for 48 h. Used as a therapeutic approach, striatopallidal inhibition should consider the risk of impairing goal-directed efficiency and behavioral desensitization. SIGNIFICANCE STATEMENT: Motivation involves a directional component that allows subjects to efficiently select the behavior that will lead to an optimal outcome and an activational component that initiates and maintains the vigor and persistence of actions. Striatal output pathways modulate motivated behavior, but it remains unknown how these pathways regulate specific components of motivation. Here, we found that the indirect pathway controls response initiation without affecting response vigor or the sensitivity to changes in the reward outcome. A specific enhancement in the activational component of motivation, however, can come at the cost of goal-directed efficiency when a sustained response is required to obtain the goal. These data should inform treatment strategies for brain disorders with impaired motivation such as schizophrenia and Parkinson's disease.

Expanding the role of striatal cholinergic interneurons and the midbrain dopamine system in appetitive instrumental conditioning.

The basal ganglia are a collection of subcortical nuclei thought to underlie a wide variety of vertebrate behavior. Although a great deal is known about the functional and physiological properties of the basal ganglia, relatively few models have been formally developed that have been tested against both behavioral and physiological data. Our previous work (Ashby FG, Crossley MJ. J Cogn Neurosci 23: 1549-1566, 2011) showed that a model grounded in the neurobiology of the basal ganglia could account for basic single-neuron recording data, as well as behavioral phenomena such as fast reacquisition that constrain models of conditioning. In this article we show that this same model accounts for a variety of appetitive instrumental conditioning phenomena, including the partial reinforcement extinction (PRE) effect, rapid and slowed reacquisition following extinction, and renewal of previously extinguished instrumental responses by environmental context cues.

Neural substrates underlying effort, time, and risk-based decision making in motivated behavior.

All mobile organisms rely on adaptive motivated behavior to overcome the challenges of living in an environment in which essential resources may be limited. A variety of influences ranging from an organism's environment, experiential history, and physiological state all influence a cost-benefit analysis which allows motivation to energize behavior and direct it toward specific goals. Here we review the substantial amount of research aimed at discovering the interconnected neural circuits which allow organisms to carry-out the cost-benefit computations which allow them to behave in adaptive ways. We specifically focus on how the brain deals with different types of costs, including effort requirements, delays to reward and payoff riskiness. An examination of this broad literature highlights the importance of the extended neural circuits which enable organisms to make decisions about these different types of costs. This involves Cortical Structures, including the Anterior Cingulate Cortex (ACC), the Orbital Frontal Cortex (OFC), the Infralimbic Cortex (IL), and prelimbic Cortex (PL), as well as the Baso-Lateral Amygdala (BLA), the Nucleus Accumbens (NAcc), the Ventral Pallidal (VP), the Sub Thalamic Nucleus (STN) among others. Some regions are involved in multiple aspects of cost-benefit computations while the involvement of other regions is restricted to information relating to specific types of costs.

Neonatal eyelid conditioning during sleep.

Using an eyelid conditioning paradigm modeled after that developed by Little, Lipsitt, and Rovee-Collier (1984), Fifer et al. (2010) demonstrated that newborn infants learn during sleep. This study examined the role of sleep state in neonatal learning. We recorded electroencephalogram (EEG), respiratory, and cardiovascular activity from sleeping full term newborn infants during delay eyelid conditioning. In the experimental group (n = 21), a tone was paired with an air puff to the eye. Consistent with Fifer et al. (2010), newborn infants reliably learned during sleep. The experimental group more than doubled EMR rates to a tone alone, while a control group (n = 17) presented with unpaired tones and puffs maintained low EMR rates. Infant learners were more likely to produce a conditioned EMR during quiet sleep compared to active sleep. Understanding the influence of sleep state on conditioned responses will inform the potential use of eyelid conditioning for early screening.

Time to rethink the neural mechanisms of learning and memory.

Most studies in the neurobiology of learning assume that the underlying learning process is a pairing - dependent change in synaptic strength that requires repeated experience of events presented in close temporal contiguity. However, much learning is rapid and does not depend on temporal contiguity, which has never been precisely defined. These points are well illustrated by studies showing that the temporal relations between events are rapidly learned- even over long delays- and that this knowledge governs the form and timing of behavior. The speed with which anticipatory responses emerge in conditioning paradigms is determined by the information that cues provide about the timing of rewards. The challenge for understanding the neurobiology of learning is to understand the mechanisms in the nervous system that encode information from even a single experience, the nature of the memory mechanisms that can encode quantities such as time, and how the brain can flexibly perform computations based on this information.

Newborn infants learn during sleep.

Newborn infants must rapidly adjust their physiology and behavior to the specific demands of the novel postnatal environment. This adaptation depends, at least in part, on the infant's ability to learn from experiences. We report here that infants exhibit learning even while asleep. Bioelectrical activity from face and scalp electrodes was recorded from neonates during an eye movement conditioning procedure in which a tone was followed by a puff of air to the eye. Sleeping newborns rapidly learned the predictive relationship between the tone and the puff. Additionally, in the latter part of training, these infants exhibited a frontally maximum positive EEG slow wave possibly reflecting memory updating. As newborns spend most of their time sleeping, the ability to learn about external stimuli in the postnatal environment during nonawake states may be crucial for rapid adaptation and infant survival. Furthermore, because eyelid conditioning reflects functional cerebellar circuitry, this method potentially offers a unique approach for early identification of infants at risk for a range of developmental disorders including autism and dyslexia.

Medial prefrontal lesions in mice impair sustained attention but spare maintenance of information in working memory.

Working memory and attention are complex cognitive functions that are disrupted in several neuropsychiatric disorders. Mouse models of such human diseases are commonly subjected to maze-based tests that can neither distinguish between these cognitive functions nor isolate specific aspects of either function. Here, we have adapted a simple visual discrimination task, and by varying only the timing of events within the same task construct, we are able to measure independently the behavioral response to increasing attentional demand and increasing length of time that information must be maintained in working memory. We determined that mPFC lesions in mice impair attention but not working memory maintenance.

Dopamine D2 receptors in nucleus accumbens cholinergic interneurons increase impulsive choice.

Impulsive choice, often characterized by excessive preference for small, short-term rewards over larger, long-term rewards, is a prominent feature of substance use and other neuropsychiatric disorders. The neural mechanisms underlying impulsive choice are not well understood, but growing evidence implicates nucleus accumbens (NAc) dopamine and its actions on dopamine D2 receptors (D2Rs). Because several NAc cell types and afferents express D2Rs, it has been difficult to determine the specific neural mechanisms linking NAc D2Rs to impulsive choice. Of these cell types, cholinergic interneurons (CINs) of the NAc, which express D2Rs, have emerged as key regulators of striatal output and local dopamine release. Despite these relevant functions, whether D2Rs expressed specifically in these neurons contribute to impulsive choice behavior is unknown. Here, we show that D2R upregulation in CINs of the mouse NAc increases impulsive choice as measured in a delay discounting task without affecting reward magnitude sensitivity or interval timing. Conversely, mice lacking D2Rs in CINs showed decreased delay discounting. Furthermore, CIN D2R manipulations did not affect probabilistic discounting, which measures a different form of impulsive choice. Together, these findings suggest that CIN D2Rs regulate impulsive decision-making involving delay costs, providing new insight into the mechanisms by which NAc dopamine influences impulsive behavior.

Temporal encoding: Relative and absolute representations of time guide behavior.

Temporal information-processing is critical for adaptive behavior and goal-directed action. It is thus crucial to understand how the temporal distance between behaviorally relevant events is encoded to guide behavior. However, research on temporal representations has yielded mixed findings as to whether organisms utilize relative versus absolute judgments of time intervals. To address this fundamental question about the timing mechanism, we tested mice in a duration discrimination procedure in which they learned to correctly categorize tones of different durations as short or long. After being trained on a pair of target intervals, the mice were transferred to conditions in which cue durations and corresponding response locations were systematically manipulated so that either the relative or absolute mapping remained constant. The findings indicate that transfer occurred most readily when relative relationships of durations and response locations were preserved. In contrast, when subjects had to re-map these relative relations, even when positive transfer initially occurred based on absolute mappings, their temporal discrimination performance was impaired, and they required extensive training to re-establish temporal control. These results demonstrate that mice can represent experienced durations both as having a certain magnitude (absolute representation) and as being shorter or longer of the two durations (an ordinal relation to other cue durations), with relational control having a more enduring influence in temporal discriminations. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Dopamine D2 receptors in nucleus accumbens cholinergic interneurons increase impulsive choice.

Impulsive choice, often characterized by excessive preference for small, short-term rewards over larger, long-term rewards, is a prominent feature of substance use and other neuropsychiatric disorders. The neural mechanisms underlying impulsive choice are not well understood, but growing evidence implicates nucleus accumbens (NAc) dopamine and its actions on dopamine D2 receptors (D2Rs). Because several NAc cell types and afferents express D2Rs, it has been difficult to determine the specific neural mechanisms linking NAc D2Rs to impulsive choice. Of these cell types, cholinergic interneurons (CINs) of the NAc, which express D2Rs, have emerged as key regulators of striatal output and local dopamine release. Despite these relevant functions, whether D2Rs expressed specifically in these neurons contribute to impulsive choice behavior is unknown. Here, we show that D2R upregulation in CINs of the mouse NAc increases impulsive choice as measured in a delay discounting task without affecting reward magnitude sensitivity or interval timing. Conversely, mice lacking D2Rs in CINs showed decreased delay discounting. Furthermore, CIN D2R manipulations did not affect probabilistic discounting, which measures a different form of impulsive choice. Together, these findings suggest that CIN D2Rs regulate impulsive decision-making involving delay costs, providing new insight into the mechanisms by which NAc dopamine influences impulsive behavior.

Temporal scaling and computing time in neural circuits: Should we stop watching the clock and look for its gears?

Timing underlies a variety of functions, from walking to perceiving causality. Neural timing models typically fall into one of two categories-"ramping" and "population-clock" theories. According to ramping models, individual neurons track time by gradually increasing or decreasing their activity as an event approaches. To time different intervals, ramping neurons adjust their slopes, ramping steeply for short intervals and vice versa. In contrast, according to "population-clock" models, multiple neurons track time as a group, and each neuron can fire nonlinearly. As each neuron changes its rate at each point in time, a distinct pattern of activity emerges across the population. To time different intervals, the brain learns the population patterns that coincide with key events. Both model categories have empirical support. However, they often differ in plausibility when applied to certain behavioral effects. Specifically, behavioral data indicate that the timing system has a rich computational capacity, allowing observers to spontaneously compute novel intervals from previously learned ones. In population-clock theories, population patterns map to time arbitrarily, making it difficult to explain how different patterns can be computationally combined. Ramping models are viewed as more plausible, assuming upstream circuits can set the slope of ramping neurons according to a given computation. Critically, recent studies suggest that neurons with nonlinear firing profiles often scale to time different intervals-compressing for shorter intervals and stretching for longer ones. This "temporal scaling" effect has led to a hybrid-theory where, like a population-clock model, population patterns encode time, yet like a ramping neuron adjusting its slope, the speed of each neuron's firing adapts to different intervals. Here, we argue that these "relative" population-clock models are as computationally plausible as ramping theories, viewing population-speed and ramp-slope adjustments as equivalent. Therefore, we view identifying these "speed-control" circuits as a key direction for evaluating how the timing system performs computations. Furthermore, temporal scaling highlights that a key distinction between different neural models is whether they propose an absolute or relative time-representation. However, we note that several behavioral studies suggest the brain processes both scales, cautioning against a dichotomy.

Dopamine encodes real-time reward availability and transitions between reward availability states on different timescales.

Optimal behavior requires interpreting environmental cues that indicate when to perform actions. Dopamine is important for learning about reward-predicting events, but its role in adapting to inhibitory cues is unclear. Here we show that when mice can earn rewards in the absence but not presence of an auditory cue, dopamine level in the ventral striatum accurately reflects reward availability in real-time over a sustained period (80 s). In addition, unpredictable transitions between different states of reward availability are accompanied by rapid (~1-2 s) dopamine transients that deflect negatively at the onset and positively at the offset of the cue. This Dopamine encoding of reward availability and transitions between reward availability states is not dependent on reward or activity evoked dopamine release, appears before mice learn the task and is sensitive to motivational state. Our findings are consistent across different techniques including electrochemical recordings and fiber photometry with genetically encoded optical sensors for calcium and dopamine.

Temporal maps and informativeness in associative learning.

Neurobiological research on learning assumes that temporal contiguity is essential for association formation, but what constitutes temporal contiguity has never been specified. We review evidence that learning depends, instead, on learning a temporal map. Temporal relations between events are encoded even from single experiences. The speed with which an anticipatory response emerges is proportional to the informativeness of the encoded relation between a predictive stimulus or event and the event it predicts. This principle yields a quantitative account of the heretofore undefined, but theoretically crucial, concept of temporal pairing, an account in quantitative accord with surprising experimental findings. The same principle explains the basic results in the cue competition literature, which motivated the Rescorla-Wagner model and most other contemporary models of associative learning. The essential feature of a memory mechanism in this account is its ability to encode quantitative information.

Oxycodone lengthens reproductions of suprasecond time intervals in human research volunteers.

Oxycodone, a popularly used opioid for treating pain, is widely abused. Other drugs of abuse have been shown to affect time perception, which, in turn, may affect sensitivity to future consequences. This may contribute to continued use. This study evaluated the effect of oxycodone on time perception in normal healthy volunteers. For this within-subject, double-blind design study, participants performed a temporal reproduction task before and after receiving placebo or oxycodone (15 mg, orally) over six outpatient sessions. Participants were first trained with feedback to reproduce three standard intervals (1.1, 2.2, and 3.3 s) in separate blocks by matching response latency from a start signal to the duration of that block's standard interval. During testing, participants were instructed to reproduce the three intervals from memory without feedback before and after drug administration. Oxycodone significantly lengthened time estimations for the two longer intervals relative to placebo. These results suggest that opioids alter temporal processing for intervals greater than 1 s, raising questions about the effect of these drugs on the valuation of future consequences.