Distinct action signals by subregions in nucleus accumbens during STOP-change performance.

Nucleus accumbens (NAc) is thought to contribute to motivated behavior by signaling the value of reward-predicting cues and the delivery of anticipated reward. NAc is subdivided into core and shell, with each region containing different populations of neurons that increase or decrease firing to rewarding events. While there are various and numerous theories of functions pertaining to these subregions and cell types, most are in the context of reward processing, with fewer considering that NAc might serve functions related to action selection more generally. We recorded from single neurons in NAc as rats of both sexes performed a STOP-change task that is commonly used to study motor control and impulsivity. In this task, rats respond quickly to a spatial cue on 80% of trials (GO) and must stop and redirect planned movement on 20% of trials (STOP). We found that the activity of reward-excited neurons signaled accurate response direction on GO, but not STOP trials, and that these neurons exhibited higher pre-cue firing after correct trials resulting in stronger firing during correct GO trials and errant STOP trials, while reward-inhibited neurons significantly represented response direction on STOP trials at the time of the instrumental response. Finally, the proportion of reward-excited to reward-inhibited neurons and the strength of pre-cue firing decreased as the electrode traversed NAc. We conclude that reward-excited cells (more common in core) promote proactive action selection, while reward-inhibited cells (more common in shell) contribute to accurate responding on STOP trials that require reactive suppression and redirection of behavior.Significant Statement The ability to appropriately adapt behavior is an important part of human cognition, and one that is disrupted by many neuropsychiatric disorders. Here we recorded from neurons in the nucleus accumbens (NAc) as rats performed a cognitive control task and found cell type- and subregion-specific firing patterns. Core and reward-excited cells track trial outcome history, proactively driving behavior to the first cue-a strategy that is appropriate for most trials. Conversely, shell and reward-inhibited neurons signal accurate response direction on trials requiring redirection of behavior. Together, these data suggest that NAc neuronal populations differentially contribute to action selection.

Psilocybin decreases neural responsiveness and increases functional connectivity while preserving pure-tone frequency selectivity in mouse auditory cortex.

Psilocybin is a serotonergic psychedelic believed to have therapeutic potential for neuropsychiatric conditions. Despite well-documented prevalence of perceptual alterations, hallucinations, and synesthesia associated with psychedelic experiences, little is known about how psilocybin affects sensory cortex or alters the activity of neurons in awake animals. To investigate, we conducted two-photon imaging experiments in auditory cortex of awake mice and collected video of free-roaming mouse behavior, both at baseline and during psilocybin treatment. In comparison with pre-dose neural activity, a 2 mg/kg ip dose of psilocybin initially increased the amplitude of neural responses to sound. Thirty minutes post-dose, behavioral activity and neural response amplitudes decreased, yet functional connectivity increased. In contrast, control mice given intraperitoneal saline injections showed no significant changes in either neural or behavioral activity across conditions. Notably, neuronal stimulus selectivity remained stable during psilocybin treatment, for both tonotopic cortical maps and single-cell pure-tone frequency tuning curves. Our results mirror similar findings regarding the effects of serotonergic psychedelics in visual cortex and suggest that psilocybin modulates the balance of intrinsic versus stimulus-driven influences on neural activity in auditory cortex.NEW & NOTEWORTHY Recent studies have shown promising therapeutic potential for psychedelics in treating neuropsychiatric conditions. Musical experience during psilocybin-assisted therapy is predictive of treatment outcome, yet little is known about how psilocybin affects auditory processing. Here, we conducted two-photon imaging experiments in auditory cortex of awake mice that received a dose of psilocybin. Our results suggest that psilocybin modulates the roles of intrinsic neural activity versus stimulus-driven influences on auditory perception.

Optogenetic Inhibition of the Orbitofrontal Cortex Disrupts Inhibitory Control during Stop-Change Performance in Male Rats.

Historically, the orbitofrontal cortex (OFC) has been implicated in a variety of behaviors ranging from reversal learning and inhibitory control to more complex representations of reward value and task space. While modern interpretations of the OFC's function have focused on a role in outcome evaluation, these cognitive processes often require an organism to inhibit a maladaptive response or strategy. Single-unit recordings from the OFC in rats performing a stop-change task show that the OFC responds strongly to STOP trials. To investigate the role that the OFC plays in stop-change performance, we expressed halorhodopsin (eNpHR3.0) in excitatory neurons in the OFC and tested rats on the stop-change task. Previous work suggests that the OFC differentiates between STOP trials based on trial sequence (i.e., gS trials: STOP trials preceded by a GO vs sS trials: STOP trials preceded by a STOP). We found that yellow light activation of the eNpHR3.0-expressing neurons significantly decreased accuracy only on STOP trials that followed GO trials (gS trials). Further, optogenetic inhibition of the OFC speeded reaction times on error trials. This suggests that the OFC plays a role in inhibitory control processes and that this role needs to be accounted for in modern interpretations of OFC function.

Rewarded Maze Training Increases Approach Behavior in Rats Through Neurogenesis-Dependent Growth of Ventral Hippocampus-Prelimbic Circuits.

Background: Learning complex navigation routes increases hippocampal volume in humans, but it is not clear whether this growth impacts behaviors outside the learning situation or what cellular mechanisms are involved. Methods: We trained rats with pharmacogenetic suppression of adult neurogenesis and littermate controls in 3 mazes over 3 weeks and tested novelty approach behavior several days after maze exposure. We then measured hippocampus and prelimbic cortex volumes using magnetic resonance imaging and assessed neuronal and astrocyte morphology. Finally, we investigated the activation and behavioral role of the ventral CA1 (vCA1)-to-prelimbic pathway using immediate-early genes and DREADDs (designer receptors exclusively activated by designer drugs). Results: Maze training led to volume increase of both the vCA1 region of the hippocampus and the prelimbic region of the neocortex compared with rats that followed fixed paths. Growth was also apparent in individual neurons and astrocytes in these 2 regions, and behavioral testing showed increased novelty approach in maze-trained rats in 2 different tests. Suppressing adult neurogenesis prevented the effects on structure and approach behavior after maze training without affecting maze learning itself. The vCA1 neurons projecting to the prelimbic area were more activated by novelty in maze-trained animals, and suppression of this pathway decreased approach behavior. Conclusions: Rewarded navigational learning experiences induce volumetric and morphologic growth in the vCA1 and prelimbic cortex and enhance activation of the circuit connecting these 2 regions. Both the structural and behavioral effects of maze training require ongoing adult neurogenesis, suggesting a role for new neurons in experience-driven increases in novelty exploration.

Pillar[6]MaxQ: A Potent Supramolecular Host for In Vivo Sequestration of Methamphetamine and Fentanyl.

Pillar[6]MaxQ (P6AS) functions as an in vivo sequestration agent for methamphetamine and fentanyl. We use 1H NMR, isothermal titration calorimetry, and molecular modelling to deduce the geometry and strength of the P6AS•drug complexes. P6AS forms tight complexes with fentanyl (Kd=9.8 nM), PCP (17.1 nM), MDMA (25.5 nM), mephedrone (52.4 nM), and methamphetamine (101 nM). P6AS has good in vitro biocompatibility according to MTS metabolic, Adenylate Kinase cell death, and hERG ion channel inhibition assays, and the Ames fluctuation test. The no observed adverse effect level for P6AS is 45 mg/kg. The hyperlocomotion of mice treated with methamphetamine (0.5 mg/kg) can be ameliorated by treatment with P6AS (35.7 mg/kg) 5-minutes later, whereas the hyperlocomotion of mice treated with fentanyl (0.1 mg/kg) can be controlled by treatment with P6AS (5 mg/kg) up to 15-minutes later. P6AS has significant potential for development as a broad spectrum in vivo sequestration agent.

Medial prefrontal cortex lesions disrupt prepotent action selection signals in dorsomedial striatum.

The ability to inhibit or adapt unwanted actions or movements is a critical feature of almost all forms of behavior. Many have attributed this ability to frontal brain areas such as the anterior cingulate cortex (ACC) and the medial prefrontal cortex (mPFC), but the exact contribution of each brain region is often debated because their functions are not examined in animals performing the same task. Recently, we have shown that ACC signals a need for cognitive control and is crucial for the adaptation of action selection signals in dorsomedial striatum (DMS) in rats performing a stop-change task. Here, we show that unlike ACC, the prelimbic region of mPFC does not disrupt the inhibition or adaption of an action plan at either the level of behavior or downstream firing in DMS. Instead, lesions to mPFC correlate with changes in DMS signals involved in action initiation and disrupt performance on GO trials while improving performance on STOP trials.

Anthracene-Walled Acyclic CB[n] Receptors: in†vitro and in†vivo Binding Properties toward Drugs of Abuse.

We report studies of the interaction of six acyclic CB[n]-type receptors toward a panel of drugs of abuse by a combination of isothermal titration calorimetry and 1 H NMR spectroscopy. Anthracene walled acyclic CB[n] host (M3) displays highest binding affinity toward methamphetamine (Kd =15 nM) and fentanyl (Kd =4 nM). Host M3 is well tolerated by Hep G2 and HEK 293 cells up to 100 µM according to MTS metabolic and adenylate kinase release assays. An in vivo maximum tolerated dose study with Swiss Webster mice showed no adverse effects at the highest dose studied (44.7 mg kg-1 ). Host M3 is not mutagenic based on the Ames fluctuation test and does not inhibit the hERG ion channel. In vivo efficacy studies showed that pretreatment of mice with M3 significantly reduces the hyperlocomotion after treatment with methamphetamine, but M3 does not function similarly when administered 30 seconds after methamphetamine.

In Vitro and In Vivo Sequestration of Methamphetamine by a Sulfated Acyclic CB[n]-Type Receptor.

We report the synthesis of two new acyclic sulfated acyclic CB[n]-type receptors (TriM0 and Me4 TetM0) and investigations of their binding properties toward a panel of drugs of abuse (1-13) by a combination of 1 H NMR spectroscopy and isothermal titration calorimetry. TetM0 is the most potent receptor with Ka ≥106  M-1 toward methamphetamine, fentanyl, MDMA and mephedrone. TetM0 is not cytotoxic toward HepG2 and HEK 293 cells below 100 µM according to MTS metabolic and adenylate kinase release assays and is well tolerated in vivo when dosed at 46 mg kg-1 . TetM0 does not inhibit the hERG ion channel and is not mutagenic based on the Ames fluctuation test. Finally, in vivo efficacy studies show that the hyperlocomotion of mice treated with methamphetamine can be greatly reduced by treatment with TetM0 up to 5 minutes later. TetM0 has potential as a broad spectrum in vivo sequestrant for drugs of abuse.

Reactive and Proactive Adaptation of Cognitive and Motor Neural Signals during Performance of a Stop-Change Task.

The ability to inhibit or suppress unwanted or inappropriate actions, is an essential component of executive function and cognitive health. The immense selective pressure placed on maintaining inhibitory control processes is exemplified by the relatively small number of instances in which these systems completely fail in the average person's daily life. Although mistakes and errors do inevitably occur, inhibitory control systems not only ensure that this number is low, but have also adapted behavioral strategies to minimize future failures. The ability of our brains to adapt our behavior and appropriately engage proper motor responses is traditionally depicted as the primary domain of frontal brain areas, despite evidence to the fact that numerous other brain areas contribute. Using the stop-signal task as a common ground for comparison, we review a large body of literature investigating inhibitory control processes across frontal, temporal, and midbrain structures, focusing on our recent work in rodents, in an effort to understand how the brain biases action selection and adapts to the experience of conflict.

Prior Cocaine Exposure Increases Firing to Immediate Reward While Attenuating Cue and Context Signals Related to Reward Value in the Insula.

The insula contributes to behavioral control and is disrupted by substance abuse, yet we know little about the neural signals underlying these functions or how they are disrupted after chronic drug self-administration. Here, male and female rats self-administered either cocaine (experimental group) or sucrose (control) for 12 consecutive days. After a 1 month withdrawal period, we recorded from insula while rats performed a previously learned reward-guided decision-making task. Cocaine-exposed rats were more sensitive to value manipulations and were faster to respond. These behavioral changes were accompanied by elevated counts of neurons in the insula that increased firing to reward. These neurons also fired more strongly at the start of long-delay trials, when a more immediate reward would be expected, and fired less strongly in anticipation of the actual delivery of delayed rewards. Although reward-related firing to immediate reward was enhanced after cocaine self-administration, reward-predicting cue and context signals were attenuated. In addition to revealing novel firing patterns unique to insula, our data suggest changes in such neural activity likely contribute to impaired decision making observed after drug use.SIGNIFICANCE STATEMENT The insula plays a clear role in drug addiction and drug-induced impairments of decision making, yet there is little understanding of its underlying neural signals. We found that chronic cocaine self-administration reduces cue and context encoding in insula while enhancing signals related to immediate reward. These changes in neural activity likely contribute to impaired decision making and impulsivity observed after drug use.

Anterior cingulate cortex and adaptive control of brain and behavior.

Research examining the functional underpinnings of anterior cingulate cortex (ACC) and its relationship to cognitive control have been described as "perennially controversial" and a "Rorschach Test" for modern neuroscience. Although there is near universal agreement that ACC is important for the adaptation of behavior, debate, despite decades of work, stems from the exact manner in which ACC goes about doing this. This chapter provides a brief overview of the various past and present theoretical arguments and research surrounding ACC function, and highlights an emerging literature of single unit ACC recordings from several species that support these theories. We will finish the chapter by focusing on our work examining the firing of single neurons in rat dorsal medial striatum (DMS) and ACC, and examining DMS's dependency on ACC to accurately signal adaptive behavioral output. Ultimately, we will conclude that ACC carries a myriad of signals (error detection, reinforcement/feedback, value, response conflict, etc.) necessary for the modulation of attention and task-relevant/irrelevant signals so that difficult decisions can be made and action plans adapted when necessary.

Prediction errors and valence: From single units to multidimensional encoding in the amygdala.

The amygdala-one of the primary structures of the limbic system-is comprised of interconnected nuclei situated within the temporal lobe. It has a well-established role in the modulation of negative affective states, as well as in fear processing. However, its vast projections with diverse brain regions-ranging from the cortex to the brainstem-are suggestive of its more complex involvement in affective or motivational aspects of cognitive processing. The amygdala can play an invaluable role in context-dependent associative learning, unsigned prediction error learning, influencing outcome selection, and multidimensional encoding. In this review, we delve into the amygdala's role in associative learning and outcome selection, emphasizing its intrinsic involvement in the appropriate context-dependent modulation of motivated behavior.

Enduring consequences of perinatal fentanyl exposure in mice.

Opioid use by pregnant women is an understudied consequence associated with the opioid epidemic, resulting in a rise in the incidence of neonatal opioid withdrawal syndrome (NOWS) and lifelong neurobehavioral deficits that result from perinatal opioid exposure. There are few preclinical models that accurately recapitulate human perinatal drug exposure and few focus on fentanyl, a potent synthetic opioid that is a leading driver of the opioid epidemic. To investigate the consequences of perinatal opioid exposure, we administered fentanyl to mouse dams in their drinking water throughout gestation and until litters were weaned at postnatal day (PD) 21. Fentanyl-exposed dams delivered smaller litters and had higher litter mortality rates compared with controls. Metrics of maternal care behavior were not affected by the treatment, nor were there differences in dams' weight or liquid consumption throughout gestation and 21 days postpartum. Twenty-four hours after weaning and drug cessation, perinatal fentanyl-exposed mice exhibited signs of spontaneous somatic withdrawal behavior and sex-specific weight fluctuations that normalized in adulthood. At adolescence (PD 35), they displayed elevated anxiety-like behaviors and decreased grooming, assayed in the elevated plus maze and sucrose splash tests. Finally, by adulthood (PD 55), they displayed impaired performance in a two-tone auditory discrimination task. Collectively, our findings suggest that perinatal fentanyl-exposed mice exhibit somatic withdrawal behavior and change into early adulthood reminiscent of humans born with NOWS.

The ever-changing OFC landscape: What neural signals in OFC can tell us about inhibitory control.

Despite decades of research on OFC function, the exact function(s) of OFC remain elusive. In recent years, 1 of the earliest hypotheses about OFC function, namely its involvement in inhibitory control, has drifted to the periphery of the functional OFC landscape in favor of theories suggesting a role for OFC in the representation of task or state space. The reasons for this drift are valid, owing in part to the development of more sensitive behavioral approaches, a clear emphasis on cross-species and cross-method comparisons, as well as the elegant integration of reinforcement learning theories. However, recent evidence recording from OFC during the performance of traditional inhibitory control tasks has found new evidence supporting a role for OFC in inhibitory control. While the extent to which these findings can be integrated into existing frameworks is in its infancy, this review seeks to highlight these findings with the goal of providing new insights into function of OFC. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

In Vitro and In Vivo Sequestration of Phencyclidine by Me4 Cucurbit[8]uril*.

We report investigations of the use of cucurbit[8]uril (CB[8]) macrocycles as an antidote to counteract the in vivo biological effects of phencyclidine. We investigate the binding of CB[8] and its derivative Me4 CB[8] toward ten drugs of abuse (3-9, 12-14) by a combination of 1 H NMR spectroscopy and isothermal titration calorimetry in phosphate buffered water. We find that the cavity of CB[8] and Me4 CB[8] are able to encapsulate the 1-amino-1-aryl-cyclohexane ring system of phencyclidine (PCP) and ketamine as well as the morphinan skeleton of morphine and hydromorphone with Kd values ≤50 nm. In vitro cytotoxicity (MTS metabolic and adenylate kinase cell death assays in HEK293 and HEPG2 cells) and in vivo maximum tolerated dose studies (Swiss Webster mice) which were performed for Me4 CB[8] indicated good tolerability. The tightest host⋅guest pair (Me4 CB[8]⋅PCP; Kd =2 nm) was advanced to in vivo efficacy studies. The results of open field tests demonstrate that pretreatment of mice with Me4 CB[8] prevents subsequent hyperlocomotion induction by PCP and also that treatment of animals previously dosed with PCP with Me4 CB[8] significantly reduces the locomotion levels.

Neural Signals in Red Nucleus during Reactive and Proactive Adjustments in Behavior.

The ability to adjust behavior is an essential component of cognitive control. Much is known about frontal and striatal processes that support cognitive control, but few studies have investigated how motor signals change during reactive and proactive adjustments in motor output. To address this, we characterized neural signals in red nucleus (RN), a brain region linked to motor control, as male and female rats performed a novel variant of the stop-signal task. We found that activity in RN represented the direction of movement and was strongly correlated with movement speed. Additionally, we found that directional movement signals were amplified on STOP trials before completion of the response and that the strength of RN signals was modulated when rats exhibited cognitive control. These results provide the first evidence that neural signals in RN integrate cognitive control signals to reshape motor outcomes reactively within trials and proactivity across them.SIGNIFICANCE STATEMENT Healthy human behavior requires the suppression or inhibition of errant or maladaptive motor responses, often called cognitive control. While much is known about how frontal brain regions facilitate cognitive control, less is known about how motor regions respond to rapid and unexpected changes in action selection. To address this, we recorded from neurons in the red nucleus, a motor region thought to be important for initiating movement in rats performing a cognitive control task. We show that red nucleus tracks motor plans and that selectivity was modulated on trials that required shifting from one motor response to another. Collectively, these findings suggest that red nucleus contributes to modulating motor behavior during cognitive control.

Anterior cingulate cortex is necessary for adaptation of action plans.

Previous research has focused on the anterior cingulate cortex (ACC) as a key brain region in the mitigation of the competition that arises from two simultaneously active signals. However, to date, no study has demonstrated that ACC is necessary for this form of behavioral flexibility, nor have any studies shown that ACC acts by modulating downstream brain regions such as the dorsal medial striatum (DMS) that encode action plans necessary for task completion. Here, we performed unilateral excitotoxic lesions of ACC while recording downstream from the ipsilateral hemisphere of DMS in rats, performing a variant of the STOP-signal task. We show that on STOP trials lesioned rats perform worse, in part due to the failure of timely directional action plans to emerge in the DMS, as well as the overrepresentation of the to-be-inhibited behavior. Collectively, our findings suggest that ACC is necessary for the mitigation of competing inputs and validates many of the existing theoretical predictions for the role of ACC in cognitive control.

A consensus guide to capturing the ability to inhibit actions and impulsive behaviors in the stop-signal task.

Response inhibition is essential for navigating everyday life. Its derailment is considered integral to numerous neurological and psychiatric disorders, and more generally, to a wide range of behavioral and health problems. Response-inhibition efficiency furthermore correlates with treatment outcome in some of these conditions. The stop-signal task is an essential tool to determine how quickly response inhibition is implemented. Despite its apparent simplicity, there are many features (ranging from task design to data analysis) that vary across studies in ways that can easily compromise the validity of the obtained results. Our goal is to facilitate a more accurate use of the stop-signal task. To this end, we provide 12 easy-to-implement consensus recommendations and point out the problems that can arise when they are not followed. Furthermore, we provide user-friendly open-source resources intended to inform statistical-power considerations, facilitate the correct implementation of the task, and assist in proper data analysis.

Single Neurons in Anterior Cingulate Cortex Signal the Need to Change Action During Performance of a Stop-change Task that Induces Response Competition.

Several human imaging studies have suggested that anterior cingulate cortex (ACC) is highly active when participants receive competing inputs, and that these signals may be important for influencing the downstream planning of actions. Despite increasing evidence from several neuroimaging studies, no study has examined ACC activity at the level of the single neuron in rodents performing similar tasks. To fill this gap, we recorded from single neurons in ACC while rats performed a stop-change task. We found higher firing on trials with competing inputs (STOP trials), and that firing rates were positively correlated with accuracy and movement speed, suggesting that when ACC was engaged, rats tended to slow down and perform better. Finally, firing was the strongest when STOP trials were preceded by GO trials and was reduced when rats adapted their behavior on trials subsequent to a STOP trial. These data provide the first evidence that activity of single neurons in ACC is elevated when 2 responses are in competition with each other when there is a need to change the course of action to obtain reward.