Clozapine N-oxide, compound 21, and JHU37160 do not influence effortful reward-seeking behavior in mice.

RATIONALE: Clozapine N-oxide (CNO) has been developed as a ligand to selectively activate designer receptors exclusively activated by designer drugs (DREADDs). However, previous studies have revealed that peripherally injected CNO is reverse-metabolized into clozapine, which, in addition to activating DREADDs, acts as an antagonist at various neurotransmitter receptors, suggesting potential off-target effects of CNO on animal physiology and behaviors. Recently, second-generation DREADD agonists compound 21 (C21) and JHU37160 (J60) have been developed, but their off-target effects are not fully understood. OBJECTIVES: The present studies assessed the effect of novel DREADD ligands on reward-seeking behavior. METHODS: We first tested the possible effect of acute i.p. injection of low-to-moderate (0.1, 0.3, 1, 3 mg/kg) of CNO, C21, and J60 on motivated reward-seeking behavior in wild-type mice. We then examined whether a high dose (10 mg/kg) of these drugs might be able to alter responding. RESULTS: Low-to-moderate doses of all drugs and a high dose of CNO or C21 did not alter operant lick responding for a reward under a progressive ratio schedule of reinforcement, in which the number of operant lick responses to obtain a reward increases after each reward collection. However, high-dose J60 resulted in a total lack of responding that was later observed in an open field arena to be due to a sedative effect. CONCLUSIONS: This study provides definitive evidence that commonly used doses of CNO, C21, and J60 have negligible off-target effects on motivated reward-seeking but urges caution when using high doses of J60 due to sedative effects.

α4-Containing GABAA Receptors on DRD2 Neurons of the Nucleus Accumbens Mediate Instrumental Responding for Conditioned Reinforcers and Its Potentiation by Cocaine.

Extrasynaptic GABAA receptors (GABAARs) composed of α4, ß, and δ subunits mediate GABAergic tonic inhibition and are potential molecular targets in the modulation of behavioral responses to natural and drug rewards. These GABAARs are highly expressed within the nucleus accumbens (NAc), where they influence the excitability of the medium spiny neurons. Here, we explore their role in modulating behavioral responses to food-conditioned cues and the behavior-potentiating effects of cocaine. α4-Subunit constitutive knock-out mice (α4-/-) showed higher rates of instrumental responding for reward-paired stimuli in a test of conditioned reinforcement (CRf). A similar effect was seen following viral knockdown of GABAAR α4 subunits within the NAc. Local infusion of the α4ßδ-GABAAR-preferring agonist THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol; Gaboxadol) into the NAc had no effect on responding when given alone but reduced cocaine potentiation of responding for conditioned reinforcers in wild-type, but not α4-/- mice. Finally, specific deletion of α4-subunits from dopamine D2, but not D1, receptor-expressing neurons (DRD2 and DRD1 neurons), mimicked the phenotype of the constitutive knockout, potentiating CRf responding, and blocking intra-accumbal THIP attenuation of cocaine-potentiated CRf responding. These data demonstrate that α4-GABAAR-mediated inhibition of DRD2 neurons reduces instrumental responding for a conditioned reinforcer and its potentiation by cocaine and emphasize the importance of GABAergic signaling within the NAc in mediating the effects of cocaine.

Dual Roles for Nucleus Accumbens Core Dopamine D1-Expressing Neurons Projecting to the Substantia Nigra Pars Reticulata in Limbic and Motor Control in Male Mice.

The nucleus accumbens (NAc) is a critical component of a limbic basal ganglia circuit that is thought to play an important role in decision-making and the processing of rewarding stimuli. As part of this circuit, dopamine D1 receptor-expressing medium spiny neurons (D1-MSNs) of the NAc core are known to send a major projection to the substantia nigra pars reticulata (SNr). However, the functional role of this SNr-projecting NAc D1-MSN (NAcD1-MSN-SNr) pathway is still largely uncharacterized. Moreover, as the SNr is thought to belong to both limbic and motor information-processing basal ganglia loops, it is possible that the NAcD1-MSN-SNr pathway may be able to influence both limbic and motor functions. In this study, we investigated the effect of optogenetic manipulation of the NAcD1-MSN-SNr pathway on reward-learning and locomotor behavior in male mice. Stimulation of the axon terminals of NAc core D1-MSNs in the SNr induced a preference for a laser-paired location, self-stimulation via a laser-paired lever, and augmented instrumental responding for a liquid reward-paired lever. Additionally, stimulation was observed to increase locomotor behavior when delivered bilaterally and induced contralateral turning behavior when delivered unilaterally. However, interestingly, inhibition of this pathway did not alter either reward-related behaviors or locomotion. These findings indicate that the NAcD1-MSN-SNr pathway is able to control both reward learning and motor behaviors.

Error-related signaling in nucleus accumbens D2 receptor-expressing neurons guides inhibition-based choice behavior in mice.

Learned associations between environmental cues and the outcomes they predict (cue-outcome associations) play a major role in behavioral control, guiding not only which responses we should perform, but also which we should inhibit, in order to achieve a specific goal. The encoding of such cue-outcome associations, as well as the performance of cue-guided choice behavior, is thought to involve dopamine D1 and D2 receptor-expressing medium spiny neurons (D1-/D2-MSNs) of the nucleus accumbens (NAc). Here, using a visual discrimination task in male mice, we assessed the role of NAc D1-/D2-MSNs in cue-guided inhibition of inappropriate responding. Cell-type specific neuronal silencing and in-vivo imaging revealed NAc D2-MSNs to contribute to inhibiting behavioral responses, with activation of NAc D2-MSNs following response errors playing an important role in optimizing future choice behavior. Our findings indicate that error-signaling by NAc D2-MSNs contributes to the ability to use environmental cues to inhibit inappropriate behavior.

Quantifying Exposure and Intra-Individual Reliability of High-Speed and Sprint Running During Sided-Games Training in Soccer Players: A Systematic Review and Meta-analysis.

BACKGROUND: Sided games (i.e., small sided, medium sided, large sided) involve tactical, technical, physical, and psychological elements and are commonly implemented in soccer training. Although soccer sided-games research is plentiful, a meta-analytical synthesis of external load exposure during sided games is lacking. OBJECTIVE: The objective of this systematic review and meta-analysis was to: (1) synthesize the evidence on high-speed and sprint running exposure induced by sided games in adult soccer players, (2) establish pooled estimates and intra-individual reliability for high-speed and sprint running exposure, and (3) explore the moderating effects of game format and playing constraints. METHODS: A literature search was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020 guidelines. Four databases (PubMed/MEDLINE, Scopus, SPORTDiscus, Web of Science Core Collection) were systematically searched up to 25 January, 2022. Eligibility criteria were adult soccer players (population); training programs incorporating sided games (intervention); game manipulations including number of players, pitch dimension, and game orientation (comparator); and high-speed, very high-speed, and sprint relative (m[Formula: see text]min-1) running distances and associated intra-individual reliability (outcome). Eligible study risk of bias was evaluated using RoBANS. Pooled estimates for high-speed and sprint running exposure, and their intra-individual reliability, along with the moderating effect of tracking device running velocity thresholds, pitch dimension (i.e., area per player), and game orientation (i.e. score or possession), were determined via a multi-level mixed-effects meta-analysis. Estimate uncertainty is presented as 95% compatibility intervals (CIs) with the likely range of relative distances in similar future studies determined via 95% prediction intervals. RESULTS: A total of 104 and 7 studies met our eligibility criteria for the main and reliability analyses, respectively. The range of relative distances covered across small-sided games, medium-sided games, and large-sided games was 14.8 m[Formula: see text]min-1 (95% CI 12.3-17.4) to 17.2 m[Formula: see text]min-1 (95% CI 13.5-20.8) for high-speed running, 2.7 m[Formula: see text]min-1 (95% CI 1.8-3.5) to 3.6 m[Formula: see text]min-1 (95% CI 2.3-4.8) for very high-speed running, and 0.2 m[Formula: see text]min-1 (95% CI 0.1-0.4) to 0.7 m[Formula: see text]min-1 (95% CI 0.5-0.9) for sprinting. Across different game formats, 95% prediction intervals showed future exposure for high-speed, very high-speed running, and sprinting to be 0-46.5 m[Formula: see text]min-1, 0-14.2 m[Formula: see text]min-1, and 0-2.6 m[Formula: see text]min-1, respectively. High-speed, very high-speed running, and sprinting showed poor reliability with a pooled coefficient of variation of 22.8% with distances being moderated by device speed thresholds, pitch dimension, and game orientation. CONCLUSIONS: This review is the first to provide a detailed synthesis of exposure and intra-individual reliability of high-speed and sprint running during soccer sided games. Our estimates, along with the moderating influence of common programming variables such as velocity thresholds, area per player, and game orientation should be considered for informed planning of small-sided games, medium-sided games, and large-sided games soccer training. CLINICAL TRIAL REGISTRATION: Open Science Framework available through https://osf.io/a4xr2/ .

Nucleus Accumbens Core Dopamine D2 Receptor-Expressing Neurons Control Reversal Learning but Not Set-Shifting in Behavioral Flexibility in Male Mice.

The ability to use environmental cues to flexibly guide responses is crucial for adaptive behavior and is thought to be controlled within a series of cortico-basal ganglia-thalamo-cortical loops. Previous evidence has indicated that different prefrontal cortical regions control dissociable aspects of behavioral flexibility, with the medial prefrontal cortex (mPFC) necessary for the ability to shift attention to a novel strategy (set-shifting) and the orbitofrontal cortex (OFC) necessary for shifting attention between learned stimulus-outcome associations (reversal learning). The nucleus accumbens (NAc) is a major downstream target of both the mPFC and the OFC; however, its role in controlling reversal learning and set-shifting abilities is still unclear. Here we investigated the contribution of the two major NAc neuronal populations, medium spiny neurons expressing either dopamine D1 or D2 receptors (D1-/D2-MSNs), in guiding reversal learning and set-shifting in an attentional set-shifting task (ASST). Persistent inhibition of neurotransmitter release from NAc D2-MSNs, but not D1-MSNs, resulted in an impaired ability for reversal learning, but not set-shifting in male mice. These findings suggest that NAc D2-MSNs play a critical role in suppressing responding toward specific learned cues that are now associated with unfavorable outcomes (i.e., in reversal stages), but not in the suppression of more general learned strategies (i.e., in set-shifting). This study provides further evidence for the anatomical separation of reversal learning and set-shifting abilities within cortico-basal ganglia-thalamo-cortical loops.

Importin α3 (KPNA3) Deficiency Augments Effortful Reward-Seeking Behavior in Mice.

Importin α3 (Gene: Kpna3, the ortholog of human Importin α4) is a member of the importin α family and participates in nucleocytoplasmic transport by forming trimeric complexes between cargo proteins and importin ß1. Evidence from human studies has indicated that single nucleotide polymorphisms (SNP) in the KPNA3 gene are associated with the occurrence of several psychiatric disorders accompanied by abnormal reward-related behavior, including schizophrenia, major depression, and substance addiction. However, the precise roles of importin α3 in controlling reward processing and motivation are still unclear. In this study, we evaluated the behavioral effects of Kpna3 knockout (KO) in mice on performance in touchscreen operant chamber-based tasks evaluating simple (fixed-ratio) and effortful (progressive-ratio) reward-seeking behaviors. While Kpna3 KO mice showed no significant differences in operant reward learning on a fixed-ratio schedule, they demonstrated significantly increased motivation (increased break point) to instrumentally respond for sucrose on a progressive-ratio schedule. We additionally measured the number of c-Fos-positive cells, a marker of neural activity, in 20 regions of the brain and identified a network of brain regions based on their interregional correlation coefficients. Network and graph-theoretic analyses suggested that Kpna3 deficiency enhanced overall interregional functional connectivity. These findings suggest the importance of Kpna3 in motivational control and indicate that Kpna3 KO mice may be an attractive line for modeling motivational abnormalities associated with several psychiatric disorders.

Effects of Importin α1/KPNA1 deletion and adolescent social isolation stress on psychiatric disorder-associated behaviors in mice.

Importin α1/KPNA1 is a member of the Importin α family widely present in the mammalian brain and has been characterized as a regulator of neuronal differentiation, synaptic functionality, and anxiety-like behavior. In humans, a de novo mutation of the KPNA1 (human Importin α5) gene has been linked with schizophrenia; however, the precise roles of KPNA1 in disorder-related behaviors are still unknown. Moreover, as recent studies have highlighted the importance of gene-environment interactions in the development of psychiatric disorders, we investigated the effects of Kpna1 deletion and social isolation stress, a paradigm that models social stress factors found in human patients, on psychiatric disorder-related behaviors in mice. Through assessment in a behavioral battery, we found that Kpna1 knockout resulted in the following behavioral phenotype: (1) decreased anxiety-like behavior in an elevated plus maze test, (2) short term memory deficits in novel object recognition test (3) impaired sensorimotor gating in a prepulse inhibition test. Importantly, exposure to social isolation stress resulted in additional behavioral abnormalities where isolated Kpna1 knockout mice exhibited: (1) impaired aversive learning and/or memory in the inhibitory avoidance test, as well as (2) increased depression-like behavior in the forced swim test. Furthermore, we investigated whether mice showed alterations in plasma levels of stress-associated signal molecules (corticosterone, cytokines, hormones, receptors), and found that Kpna1 knockout significantly altered levels of corticosterone and LIX (CXCL5). Moreover, significant decreases in the level of prolactin were found in all groups except for group-housed wild type mice. Our findings demonstrate that Kpna1 deletion can trigger widespread behavioral abnormalities associated with psychiatric disorders, some of which were further exacerbated by exposure to adolescent social isolation. The use of Kpna1 knockout mice as a model for psychiatric disorders may show promise for further investigation of gene-environment interactions involved in the pathogenesis of psychiatric disorders.

Natural and Artificial Intelligence: A brief introduction to the interplay between AI and neuroscience research.

Neuroscience and artificial intelligence (AI) share a long history of collaboration. Advances in neuroscience, alongside huge leaps in computer processing power over the last few decades, have given rise to a new generation of in silico neural networks inspired by the architecture of the brain. These AI systems are now capable of many of the advanced perceptual and cognitive abilities of biological systems, including object recognition and decision making. Moreover, AI is now increasingly being employed as a tool for neuroscience research and is transforming our understanding of brain functions. In particular, deep learning has been used to model how convolutional layers and recurrent connections in the brain's cerebral cortex control important functions, including visual processing, memory, and motor control. Excitingly, the use of neuroscience-inspired AI also holds great promise for understanding how changes in brain networks result in psychopathologies, and could even be utilized in treatment regimes. Here we discuss recent advancements in four areas in which the relationship between neuroscience and AI has led to major advancements in the field; (1) AI models of working memory, (2) AI visual processing, (3) AI analysis of big neuroscience datasets, and (4) computational psychiatry.

Parallel and hierarchical neural mechanisms for adaptive and predictive behavioral control.

Our brain can be recognized as a network of largely hierarchically organized neural circuits that operate to control specific functions, but when acting in parallel, enable the performance of complex and simultaneous behaviors. Indeed, many of our daily actions require concurrent information processing in sensorimotor, associative, and limbic circuits that are dynamically and hierarchically modulated by sensory information and previous learning. This organization of information processing in biological organisms has served as a major inspiration for artificial intelligence and has helped to create in silico systems capable of matching or even outperforming humans in several specific tasks, including visual recognition and strategy-based games. However, the development of human-like robots that are able to move as quickly as humans and respond flexibly in various situations remains a major challenge and indicates an area where further use of parallel and hierarchical architectures may hold promise. In this article we review several important neural and behavioral mechanisms organizing hierarchical and predictive processing for the acquisition and realization of flexible behavioral control. Then, inspired by the organizational features of brain circuits, we introduce a multi-timescale parallel and hierarchical learning framework for the realization of versatile and agile movement in humanoid robots.

The school playground environment as a driver of primary school children's physical activity behaviour: A direct observation case study.

The school playground can promote PA for large numbers of children. This study identifies areas of the playground that children visited at break-times, the decisions according to gender and the influence of contextual and environmental variables on PA levels. The playground of a culturally diverse primary school was observed during morning break-times and lunchtimes. Counts of sedentary, LPA, and MVPA episodes, and the contexts in which they occurred were recorded using the system for observing play and leisure in youth (SOPLAY). Ball sports areas had higher counts of boys (mean ± SD; 9.9 ± 4.8) compared to girls (2.0 ± 3.5); areas promoting climbing and social interaction had higher counts of girls (7.9 ± 7.2) compared to boys (3.5 ± 2.9). The proportion of MVPA episodes during break-times was 34% ± 26%. Areas of the playground with organised activities had 2.70 (95%CI: 1.87 to 3.91) times higher MVPA counts than areas "not organised". Areas with "supervision" were associated with higher MVPA counts (1.34; 1.18 to 1.53) compared with "not supervised" areas. Organisation and supervision might influence PA choices and PA levels of children in the primary school playground. Further investigation is required to explore different playgrounds settings, and context and gender preferences.

Effect of Sand on Knee Load During a Single-Leg Jump Task: Implications for Injury Prevention and Rehabilitation Programs.

Richardson, MC, Murphy, S, Macpherson, T, English, B, Spears, I, and Chesterton, P. Effect of sand on knee load during a single-leg jump task: implications for injury prevention and rehabilitation programs. J Strength Cond Res 34(11): 3164-3172, 2020-The purpose of the study was to determine potential differences in landing strategies and subsequent joint loads at the knee (knee abduction moment [KAM], anterior-posterior [AP] tibial translation, and total knee shear force) when jumping onto sand and firm ground from both a level surface and a 30-cm height. Firm ground would act as the control for the study. Seventeen subjects (age: 23.6 ± 3.7 years; body mass: 67.7 ± 10.3 kg; height: 168.5 ± 7.4 cm) performed 3 single-leg jumps on their dominant leg for each of the 4 conditions tested (ground level, sand level, ground height, and sand height). A repeated-measures design investigated the effect of sand on KAM, AP tibial translation, and total knee shear force. Data were analyzed using magnitude-based inferences and presented as percentage change with 90% confidence limits. Results indicated that sand had a clear beneficial effect on KAM, which was possibly moderate during a drop jump (30 cm) and possibly small from a level jump. Sand also had a possibly moderate beneficial effect on AP tibial translation from a level jump. The effect of sand on total knee shear force was unclear. These results suggest that sand may provide a safer alternative to firm ground when performing jump tasks commonly used in anterior cruciate ligament and patellofemoral joint injury prevention and rehabilitation programs. Sand may also allow for an accelerated rehabilitation program because jumping activities could potentially be implemented more safely at an earlier stage in the process.

Systematic Reductions in Differential Ratings of Perceived Exertion Across a 2-Week Repeated-Sprint-Training Intervention That Improved Soccer Players' High-Speed-Running Abilities.

PURPOSE: To quantify changes in differential ratings of perceived exertion (dRPE) across a 2-wk repeated-sprint-training intervention that improved high-intensity intermittent-running ability and linear speed of semiprofessional soccer players. METHODS: Thirteen players completed 3 (sessions 1-3) or 4 (sessions 4-6) sets of 7 sprints (group 1 [n = 7]: 30-m straight; group 2 [n = 6]: 2 × 10-m shuttle), with 20 s and 4 min of recovery between sprints and sets, respectively. Postset perceptions of breathlessness (RPE-B) and leg-muscle exertion (RPE-L) were rated using the CR100 scale. RESULTS: Overall, RPE-B (mean [SD]: 46 [13] arbitrary units [AU], "hard") was most likely higher than RPE-L (39 [13] AU, "somewhat hard," mean difference: 8 AU; 90% confidence limits [CLs]: ±2). Set-to-set increases in dRPE (in AU; 90% CL: approximately ±2) were large in session 1 (RPE-B: 15; RPE-L: 14), moderate in sessions 2-5 (RPE-B: 7-10; RPE-L: 7-8), and small (RPE-B: 6) to moderate (RPE-L: 7) in session 6. Across the intervention, RPE-B reduced moderately in sets 3 (-13; 90% CL: ±4) and 4 (-12; 90% CL: ±12) and RPE-L reduced by a small magnitude in set 3 (-5; 90% CL: ±6). The set 4 change in RPE-L was unclear (-11; 90% CL: ±13). CONCLUSIONS: The authors observed systematic intrasession and intersession changes in dRPE across a 2-wk repeated-sprint-training intervention, with a fixed prescription of external load that improved semiprofessional soccer players' high-speed-running abilities. These findings could support dRPE as a measure of internal load and highlight its usefulness in evaluating repeated-sprint-training dose-response.

Neurotransmission through dopamine D1 receptors is required for aversive memory formation and Arc activation in the cerebral cortex.

Dopaminergic neurotransmission is considered to play an important role not only in reward-based learning, but also in aversive learning. Here, we investigated the role of dopaminergic neurotransmission via dopamine D1 receptors (D1Rs) in aversive memory formation in a passive avoidance test using D1R knockdown (KD) mice, in which the expression of D1Rs can conditionally and reversibly be controlled by doxycycline (Dox) treatment. We also performed whole-brain imaging after aversive footshock stimulation in activity-regulated cytoskeleton protein (Arc)-dVenus D1RKD mice, which were crossbred from Arc-dVenus transgenic mice and D1RKD mice, to examine the distribution of Arc-controlled dVenus expression in the hippocampus and cerebral cortex during aversive memory formation. Knockdown of D1R expression following Dox treatment resulted in impaired performance in the passive avoidance test and was associated with a decrease in dVenus expression in the cerebral cortex (visual, somatosensory, and motor cortices), but not the hippocampus, compared with control mice without Dox treatment. These findings indicate that D1R-mediated dopaminergic transmission is critical for aversive memory formation, specifically by influencing Arc expression in the cerebral cortex.

Nucleus accumbens pathways control cell-specific gene expression in the medial prefrontal cortex.

The medial prefrontal cortex (mPFC) is a critical component of a cortico-basal ganglia-thalamo-cortical loop regulating limbic and cognitive functions. Within this circuit, two distinct nucleus accumbens (NAc) output neuron types, dopamine D1 or D2 receptor-expressing neurons, dynamically control the flow of information through basal ganglia nuclei that eventually project back to the mPFC to complete the loop. Thus, chronic dysfunction of the NAc may result in mPFC transcriptomal changes, which in turn contribute to disease conditions associated with the mPFC and basal ganglia. Here, we used RNA sequencing to analyse differentially expressed genes (DEGs) in the mPFC following a reversible neurotransmission blocking technique in D1 or D2 receptor-expressing NAc neurons, respectively (D1-RNB, or D2-RNB). Gene Set Enrichment Analysis revealed that gene sets of layer 5b and 6 pyramidal neurons were enriched in DEGs of the mPFC downregulated in both NAc D1- and D2-RNB mice. In contrast, gene sets of layer 5a pyramidal neurons were enriched in upregulated DEGs of the mPFC in D1-RNB mice, and downregulated DEGs of the mPFC in D2-RNB mice. These findings reveal for the first time that NAc output pathways play an important role in controlling mPFC gene expression.

Adolescent psychosocial stress enhances sensitization to cocaine exposure in genetically vulnerable mice.

Development of drug addictive behaviors is modulated by both genetic and environmental risk factors. However, the molecular mechanisms remain unknown. To address the role of adolescent stress in the development of drug addiction, we combined a transgenic mouse model in which a putative dominant-negative form of DISC1 under expressional control of the prion protein promoter is used as a genetic risk factor and adolescent social isolation stress as a gene-environmental interaction (GXE). Repeated cocaine exposure induced greater locomotion in the GXE group than in the other groups. In a conditioned place preference (CPP) test, GXE mice exhibited a significant place preference to the cocaine-conditioned area compared with the other groups. In the nucleus accumbens (NAc) of GXE mice, we found increased enzyme activity of phosphodiesterase-4 (PDE4), predominantly located in NAc D2-receptor-expressing neurons, and enhanced effects of the PDE4 inhibitor rolipram, but not the D1 agonist SKF81297, on the phosphorylation of DARPP-32 and GluA1 at PKA sites. Rolipram injection before cocaine exposure completely inhibited cocaine-induced hyperlocomotion and CPP in the GXE group. These results indicate that GXE enhances sensitivity to repeated cocaine exposure via an increase in PDE4 activity in NAc D2-recptor-expressing neurons, leading to the development of cocaine addictive behaviors.

Using differential ratings of perceived exertion to assess agreement between coach and player perceptions of soccer training intensity: An exploratory investigation.

We aimed to assess the coach-player agreement of subjective soccer training loads via differential ratings of perceived exertion (dRPE). The coach initially underwent quantifiable familiarisation (blackness test) with the Borg CR100 scale. Data were collected from 16 semi-professional soccer players across seven consecutive training sessions. For the measurement of subjective training load, the coach and players provided dRPE (CR100) for legs (RPE-L), breathlessness (RPE-B) and technical exertion (RPE-T). Coach-prescribed dRPE were recorded prior to training, with coach observed and player reported dRPE collected post training. Statistical equivalence bounds for agreement between coach (prescribed and observed) and player reported dRPE scores were 4 arbitrary units on the CR100 and we used a probability outcome of likely (≥75%) to infer realistic equivalence. Following three familiarisation sessions, the coach improved their blackness test score from 39% to 83%. Coach observed and player reported RPE-T scores were likely equivalent, with all other comparisons not realistically equivalent. Since training prescription is coach-led, our data highlight the importance of accurate internal load measurement and feedback in soccer. The improved accuracy and precision of coach intensity estimation after three attempts at the blackness test suggests that this method could be worthwhile to researchers and practitioners employing dRPE.

Preproenkephalin-expressing ventral pallidal neurons control inhibitory avoidance learning.

The ventral pallidum (VP) is a critical component of the basal ganglia neurocircuitry regulating learning and decision making; however, its precise role in controlling associative learning of environmental stimuli conditioned to appetitive or aversive outcomes is still unclear. Here, we investigated the expression of preproenkephalin, a polypeptide hormone previously shown to be expressed in nucleus accumbens neurons controlling aversive learning, within GABAergic and glutamatergic VP neurons. Next, we explored the behavioral consequences of chemicogenetic inhibition or excitation of preproenkephalin-expressing VP neurons on associative learning of reward- or aversion-paired stimuli in autoshaping and inhibitory avoidance tasks, respectively. We reveal for the first time that preproenkephalin is expressed predominantly in GABAergic rather than glutamatergic VP neurons, and that excitation of these preproenkephalin-expressing VP neurons was sufficient to impair inhibitory avoidance learning. These findings indicate the necessity for inhibition of preproenkephalin-expressing VP neurons for avoidance learning, and suggest these neurons as a potential therapeutic target for psychiatric disorders associated with maladaptive aversive learning.