Neuroscience

Electrical transmission is mediated by intercellular channels that cluster into structures known as gap junctions (GJ). In vertebrates, GJ channels are encoded by the gene family of connexin (Cx) proteins that assemble as hexamers, termed hemichannels, in the pre- and postsynaptic membranes, and that subsequently dock to form GJ channels. Auditory contacts on the fish Mauthner cells serve as model to study the properties and organization of vertebrate electrical synapses. Electrical transmission at these synapses is mediated by multiple co-existing GJs at which the presence of intercellular channels is regulated by a molecular scaffold. Zebrafish contain four homologs of the neuronal Cx36: Cx35.5 and Cx35.1 (gjd2a and b, respectively), and Cx34.1 and Cx34.7 (gjd1a and b). Cx mutations suggested that GJs are formed by heterotypic channels made of presynaptic Cx35.5 and postsynaptic Cx34.1. Using transgenic fish in which Cxs were tagged, we found that a second Cx, Cx34.7, is present together with Cx34.1 on the postsynaptic side at some but not all GJs at these terminals. When exogenously expressed, both Cx34.1 and Cx34.7 formed heterotypic functional channels with Cx35.5, each with substantially different voltage-dependent properties, indicating they can serve differential functions. However, we previously demonstrated that electrical transmission is lost in Cx34.1 but not Cx34.7 null mutants, suggesting that Cx34.7 cannot compensate for the loss of Cx34, despite the intrinsic ability of Cx34.1 and Cx34.7 to create functional channels. The findings reveal an unanticipated functional organization in the electrical synapse, where Cx34.1 is obligatory and Cx34.7 accessory, roles that appear to be defined by the postsynaptic molecular scaffold, with two postsynaptic Cxs possibly assembling under specific functional contexts. Thus, our results indicate that electrical synapses share an organizational motif with chemical synapses, akin to how they combine postsynaptic receptor types to modify synaptic function.

The amplitude of motor-evoked potentials (MEPs) elicited using transcranial magnetic stimulation (TMS) has been shown to decrease in the short interval prior to response initiation. The cause of this premovement MEP suppression is currently unclear and has been attributed to various processes such as preparation-related inhibition preventing the premature release of planned action or increasing signal-to-noise ratio to facilitate rapid response initiation. The present study explored whether the decrease in MEP amplitude is affected by the task requirements, using reaction time (RT) paradigms that differ in the timeline of preparation and initiation of a motor response. Participants completed simple RT (SRT), choice RT (CRT), and go/no-go (GNG) tasks, while TMS was applied at various times between the warning signal and go-signal. It was hypothesized that if MEP suppression relates to preparation level, the greatest suppression would be observed during the SRT and GNG tasks, as these paradigms encourage advance preparation and response inhibition. Conversely, if the reduction in corticospinal excitability is associated with facilitating response initiation processes, then suppression would be expected for all tasks, including the CRT paradigm in which preparation does not occur until presentation of the go-signal. Results showed MEP amplitudes decreased for all tasks as the go-signal approached; however, both the SRT and GNG had significantly greater MEP suppression 50 ms prior to, and coincident with the go-signal. These results indicate that the nature and origin of the suppression is likely multifactorial and relates to both preparatory and initiation-related processes, with the timeline and magnitude of suppression dependent on the nature of the task being executed. Impact StatementTranscranial magnetic stimulation was used to elicit motor-evoked potentials to examine the timeline of corticospinal activation during the instructed delay period for choice, simple and go/no-go reaction time tasks. For all tasks, corticospinal excitability was initially elevated compared to baseline, followed by a similar magnitude of early suppression. However, just prior to the go-signal, those tasks that allowed advance preparation showed additional suppression, providing novel information linking pre-movement corticospinal suppression to preparatory and inhibition processes.

Hippocampal area CA2 has recently emerged as a critical region for social recognition memory. Furthermore, this understudied region has been implicated in psychiatric diseases and neurodegenerative diseases. There has been accumulating evidence indicating that the pyramidal neurons (PNs) in area CA2 exhibit functional specializations that correlate with somatic position in stratum pyramidale (sp). In this study, we investigated the morphological differences in dendritic architecture of CA2 PNs with a focus on the radial gradient, i.e., along the deep-superficial axis of the sp. We conducted a comprehensive morphological analysis including Sholl intersection profiles, branching order distributions, root angle distributions, and dendritic cable lengths. We found that CA2 PNs have fewer oblique dendrites and a larger number of tuft-like dendrites as compared to CA1 PNs. Furthermore, within the CA2 population, we found that many of the dendritic structural features gradually changed along the radial axis from deep to superficial somatic location, indicating a continuum of dendritic morphology rather than two sharply defined subtypes of pyramidal neurons. This morphological characterization may serve as a starting point to better understand the corresponding functional organization of CA2. The gradual difference between deeper and superficial CA2 PNs suggests a continuum of their computational capabilities beyond two binary functional classes. In briefUsing several methods, we examine the dendritic morphology of over 130 CA2 and CA1 pyramidal neurons and find that many properties such as the cable length and terminal numbers of the dendritic arbors vary as a with the location of the soma in the pyramidal layer. HighlightsO_LIWe use scholl analysis, graph theory and machine learning techniques to quantify the different dendritic morphologies of CA2 pyramidal neurons. C_LIO_LIMany properties of CA2 pyramidal neuron apical dendrites vary as a function of somatic location in the pyramidal layer. C_LIO_LIMore superficial CA2 pyramidal neurons have longer oblique apical dendrites, and shorter tuft dendrites. C_LI

Homeostatic plasticity of intrinsic excitability (IE) in the visual system has been essentially shown at the cortical level but whether thalamic nuclei also express homeostatic plasticity of IE is unknown. We show here that 4 days of monocular deprivation (MD) at eye opening induces a homeostatic change in IE in dorsal lateral geniculate nucleus (dLGN) neurons. Neurons recorded in the dLGN region activated by the deprived eye are more excitable than neurons recorded in the dLGN region activated by the open eye. No significant changes were observed following 7 days of MD, however. Enhanced excitability in neurons from the deprived side after 4 days of MD was associated with a reduced Kv1-dependent LTP-IE, a smaller voltage ramp, and a reduced inter-spike interval, suggesting that Kv1 channels are down-regulated in deprived dLGN neurons. Furthermore, the ankyrin G signal of the axon initial segment was larger in deprived dLGN neurons compared with open ones, indicating that Nav1 channel number also undergoes homeostatic regulation, and Kv1.1 channel signals were lower in deprived neurons compared to open ones. In addition, electrical coupling was found to be strengthened in neurons displaying enhanced IE following either brief (4 days) or long (10 days) MD. These results suggest that homeostatic and Hebbian plasticity in the dLGN share common expression mechanisms involving the regulation of Kv1 channels, Nav1 channels and electrical coupling between relay neurons.

Glutamatergic neuronal synapses in the mouse neocortex mature during the first two months after birth. A key event during synaptic maturation is a change in short-term synaptic plasticity (STP), i.e. a switch from strong synaptic depression to a weaker depression or even facilitation. Glutamatergic pyramidal neurons located in the cortical layers II/III, layer V, and layer VI project axons through the corpus callosum where they release glutamate along their shafts and form glutamatergic synapses with oligodendrocyte precursor cells (OPCs). Here, we used single-cell electrophysiological recordings in brain slices to investigate synaptic plasticity at neuron-OPC synapses along axonal shafts in the white matter, and applied computation approaches to pinpoint the mechanisms of this plasticity. We found that during postnatal development of mice, there is a switch from short-term synaptic depression to short-term synaptic facilitation at glutamatergic neuron-OPC synapses in the corpus callosum. Synaptic delay of phasic neuron-OPC excitatory postsynaptic current shortens, and the amount of asynchronous release at neuron-OPC synapses decrease as animals mature, indicating that glutamate release becomes more synchronized. Our computational modelling suggests that both pre- and postsynaptic changes may contribute to the functional development and changes of plasticity at neuron-OPC synapses in the white matter. Taking together, our findings indicate that synaptic release machineries located at different sites along the same axon (i.e. axonal shaft in the white matter vs synaptic boutons in the grey matter) mature in a very similar fashion, STP occurs at both synaptic sites, and STP dynamics represent an important event during brain maturation.

Voltage-gated sodium channels (VGSCs) are conventionally described as heterotrimers composed of one alpha and two beta subunits. However, the patterns of co-expression of alpha- and beta-subunits in neurons remain unclear. In the present study, we report that alpha- (Nav1.1, Nav1.2, and Nav1.6) and beta- (beta-1 and beta-2) subunits are densely expressed in axon initial segments (AISs) of neurons in the neocortex, hippocampus and cerebellum at postnatal days 14-15 (P14-15) and 8-9 weeks (8-9W). These distributions are largely unique and partially overlapping among brain regions. Notably, in the neocortex and hippocampus, AISs of presumptive parvalbumin-positive inhibitory neurons are positive for Nav1.1 and beta-1, whereas those of excitatory ones are positive for Nav1.2 and beta-2. Similarly, AISs of cerebellar basket cells, which are inhibitory neurons, are positive for Nav1.1 and beta-1, whereas those of granule cells, which are excitatory neurons, are positive for Nav1.2 and beta-2. Nav1.6 is expressed in many of these neurons. Some subunits exhibited distinct distribution patterns at the two postnatal stages analyzed, possibly because of their developmental changes of subcellular localizations. Taken together, these results indicate that combinations of VGSC subunits are largely unique among different neuronal subpopulations. These findings provide a useful reference for understanding the distribution and interactions of VGSC subunits in the brain.

The addictive properties of opioids are due in part to these drugs ability to alter ventral tegmental area (VTA) activity via activation of mu opioid receptors (MORs) on local and distal inputs. Prior studies have identified numerous opioid-modulated afferents to the VTA, some of which show differing levels of functional modulation by opioids, but the degree to which this parallels differences in receptor expression is not known. Hence, we used retrograde labeling combined with RNAscope to examine oprm1 mRNA expression in VTA-projecting afferents arising from a variety of distal brain regions. Because opioids are thought to be particularly influential on GABAergic afferents to the VTA, we also examined colocalization of oprm1 with GABAergic markers in VTA-projecting neurons. Interestingly, we found that oprm1 mRNA is present in both GABAergic and non-GABAergic VTA-projecting neurons. However, many (though not all) GABAergic afferents expressed higher levels of oprm1 compared to most non-GABAergic afferents (especially those arising from the cortex). These results complement previous anatomical studies that had examined oprm1 expression in these regions but in a non-quantitative way and without regard to their efferent targets. Our findings encourage future work to examine the functional implications of MOR sensitivity within these afferent pathways.

Reward can enhance motor performance. However, its potential to counteract motor fatigability, a reduction in motor performance during sustained movements, remains underinvestigated. This could be particularly relevant in neurological conditions such as multiple sclerosis, where increased motor fatigability is a prominent symptom. One form of motor fatigability is motor slowing, a decline in movement speed over time evoked by fast, repetitive movements. In this study, we investigated whether the possibility to earn reward attenuates motor slowing, and examined associated changes in muscle activity and pupil size, a putative marker of physical effort. Participants performed a wrist tapping task at maximal voluntary speed with or without the possibility of earning a reward. We found that wrist tapping induced motor slowing and that slowing was significantly reduced by reward. Over time, tapping became more costly as indicated by higher muscle activity and coactivation per tap. This was accompanied by a sustained pupil dilation, which could not solely be explained by tapping speed. These findings suggest that, rather than restoring efficient motor control, reward attenuates motor slowing by allowing participants to access a performance reserve and invest more resources into the task, reflected by increased muscle activation per tap and sustained pupil dilation.

The lateral reticular nucleus (LRN) is thought to contribute to skilled forelimb control but its specific contributions to reaching and grasping remain unclear. In this paper, we examine skilled reaching in intact adult female Long-Evans rats after bilateral LRN ablation through single-pellet reaching tasks. Tasks were analyzed using sensitive quantitative kinematic analyses and qualitative behavioral scoring. Overall, limb transport was largely preserved after ablation, with results appearing in temporally restricted differences. The clearest deficits emerged in pellet-directed endpoint control. LRN-ablated animals showed broad variability in end-point covariance, endpoint spread, and increased trial-to-trial variability, indicating that the movement became less precise and less consistent. These effects were more consistent than any single spatial difference seen, suggesting that ablation of the LRN impairs movement refinement rather than inducing a simple directional bias, although the paw height during the reach was significantly effected. Reach duration also changed, but this temporal difference emerged later and was less prominent. Our results suggest that the LRN acts as an important contributor to endpoint stabilization and reach timing during skilled forelimb behavior.

Most syntactic approaches converge on the fact that Tense and Agreement are two different functional categories, although there is less agreement on their exact representation and relative hierarchical order. Cross-linguistic agrammatic data seems to support the difference between Tense and Agreement, with patterns of dissociation reported from agrammatism between them, in which Tense is generally more impaired than Agreement. To examine whether there is evidence for such a dissociation of tense and agreement processing in neurotypical individuals, the present study employed Event-Related brain Potentials (ERPs) to study the real-time comprehension of Modern Standard Arabic sentences. Critical stimulus sentences were of the form Temporal Adverb-Subject-Verb-PP, in which the intransitive verb was in either the past or future tense, and was preceded by a singular or plural subject and an adverb indicating past or future tense. The subject nouns were all human and either masculine or feminine. The verbs either agreed with the subject noun or presented a person, number or gender agreement violation. They also either agreed or showed a mismatch with the temporal frame of the adverb, the latter being a tense violation. Results at the verb showed that both tense and agreement violations yielded a biphasic N400 - P600 effect. We discuss these results in light of previous ERP findings and conclude that despite the putative configurational differences between Tense and Agreement, the processing of the two categories in Arabic may deploy the same underlying cognitive mechanisms.

The aim of this study was to improve our understanding of muscle contractions in the arm as a function of hand orientation for grasp. While there have been several reports on arm kinematics for reach and grasp movements, little has been done at the muscular level. To this end, we analyzed the modulation of shoulder, elbow and hand muscles for a reach and grasp task involving a target in either horizontal or vertical orientation. We hypothesized that unlike what has been observed for kinematics, at the muscular level we would see less correlation between the three muscle groups. A decoding approach with Machine Learning revealed adaptation patterns that were not visible using classical methods. Reach-and-grasp has traditionally been treated as being made of two components - the reach and the grasp components. Our dynamic decoding approach revealed a more complex picture with very different dynamics in the shoulder and elbow muscle groups during reach. All muscle groups showed peak capacity for predicting hand orientation before the start of grasp and followed the ubiquitous proximo-distal organization. The patterns of muscular modulation for hand orientation were strongly perturbed by the eyes closed and slow movement conditions, potentially decreasing the available degrees of freedom for adaptation.

The paraventricular nucleus (PVN) plays a central role in neuroendocrine and autonomic regulation, including male sexual behavior. Dopaminergic and nitrergic signaling within the PVN are functionally linked, but their cellular relationship remains unclear. We examined the colocalization of dopamine D1 and D2 receptors with neuronal nitric oxide synthase (nNOS) in adult male rats using double-label immunohistochemistry. nNOS-immunoreactive neurons were widely distributed in PVN, with subsets co-expressing D1R or D2R. Approximately 45% of nNOS-positive neurons expressed dopaminergic receptors. These findings provide structural evidence for dopaminergic-nitrergic interaction in the PVN and support the possibility of direct dopaminergic modulation of nitrergic neurons.

Dorsal (PMd) and ventral (PMv) premotor cortices can modulate contralateral primary motor cortex (M1) excitability, but their distinct interhemispheric influence via transcranial magnetic stimulation (TMS) remains unclear. Single-pulse TMS over PMd, PMv and M1 assessed transcallosal inhibition via the ipsilateral silent period (iSP). Dual-site TMS examined short-(10 ms inter-stimulus interval [ISI]), long-(50 ms ISI) and non-callosal-(0 ms ISI) interhemispheric inhibition (IHI). An iSP was elicited from PMd, PMv, and M1, with distinctly evoked iSP parameters. The iSP magnitude was greatest from M1, followed by PMd and then PMv, while iSP duration was greatest for M1 and showed no differences between PMd and PMv. Dual-site TMS revealed that PMd and M1 inhibited contralateral M1 excitability across all ISIs, while PMv showed inhibition at 0-and 50-ms ISIs. PMd and M1 demonstrated greater short-IHI compared to PMv, all demonstrating similar long-IHI, and PMd demonstrating greater non-callosal-IHI than M1. PMv displayed distinct IHI across ISIs, PMd showed differences across most ISIs and M1 demonstrated the fewest differences across ISIs. Longer iSP duration related to greater long-IHI magnitude elicited from PMd and PMv. Our findings demonstrate differential IHI from PMd and PMv on contralateral M1, which may inform neuromodulation strategies in rehabilitation contexts.

Dopamine (DA) neurons of the midbrain project throughout the striatum, including the nucleus accumbens core (NAc) and are thought to co-release ATP with DA from vesicles. The mechanisms of evoked NAc ATP release and clearance and their relationship to exocytotic DA transmission are largely unexplored and the focus of the present work. Using fast scan cyclic voltammetry (FSCV), we measured simultaneous ATP and DA transmission in response to pharmacological manipulations of release and reuptake cellular machinery. ATP transmission is tightly coupled to that of DA, though ATP release concentrations are typically smaller. Manipulations that increase DA transmission (increased release via 4-aminopyridine Kv channel blockade or decreased uptake via cocaine) also increase ATP transmission, though to a smaller extent. Blocking DA vesicular packaging (reserpine) or action potentials (lidocaine), results in attenuated DA and ATP release. Interestingly, reserpine or lidocaine can result in completely abolished DA release, but not a complete prevention in ATP release, suggesting a secondary source for ATP transmission thats not dependent on DA terminals. Both transmitters were reduced to a similar extent following nAChR blockade, demonstrating that nAChR activation regulates ATP in addition to DA. Surprisingly, cocaine inhibition of DATs reduced clearance for both ATP and DA, which correlated with one another when cocaine concentration was highest. There was also a strong relationship between the effect of cocaine on release of ATP and DA. As the first FSCV study to examine evoked NAc ATP release, this paper bridges prior work to confirm the strong association between ATP and DA in the mesolimbic circuit and identifies unexpected overlap in mechanisms regulating their transmission. Our results contribute novel evidence of both vesicular and non-vesicular ATP release in the NAc and demonstrate that extracellular ATP is a modulator of DA terminal function.

Arrestins play key role in desensitization of G protein-coupled receptors. Direct signaling role of arrestins has also been documented. Two ubiquitously expressed arrestin isoforms, arrestin-2 and -3 (Arr3), perform similarly in receptor desensitization and share many signaling functions, enabling them to substitute for one another. However, certain signaling roles are specific to each isoform. Mice lacking Arr3 (A3KO) show blunted acute responsiveness to the locomotor stimulatory effect of amphetamine (AMPH). Here we demonstrate that AMPH- and cocaine-induced locomotion of A3KO mice is significantly reduced. This loss-of-function phenotype suggests that Arr3-mediated signaling contributes to the effect. Virus-driven expression of Arr3 in caudate-putamen of A3KO and wild type mice suppressed AMPH-induced locomotion. In contrast, restoration of Arr3 in nucleus accumbens rescued locomotor response. Thus, in caudate-putamen Arr3 participates in the desensitization of dopamine receptors, whereas Arr3-dependent signaling in nucleus accumbens underlies the molecular mechanism of the locomotor response and sensitization. Using monofunctional Arr3-derived peptides, we showed that in the nucleus accumbens Arr3 promoted drug-induced locomotor responses via facilitation of JNK3 activation.

Sympathetic preganglionic neurons (SPNs) distribute signals widely across paravertebral ganglia, yet the reliability of spike propagation along their predominantly unmyelinated axons remains poorly defined. We examined temperature- and activity-dependent modulation of SPN axonal conduction using an ex vivo adult mouse thoracic sympathetic chain preparation. Population compound action potentials (CAPs) were evoked by supramaximal stimulation of T10 ventral roots and recorded from branching axons in interganglionic compared to unbranching axons in the splanchnic nerve. At physiological temperature (36{degrees}C), scaled CAP magnitude was reduced by [~]50% relative to 22{degrees}C, with preferential loss of slower-conducting axonal components. These reductions are consistent with substantial temperature-dependent decreases in effective axonal recruitment, likely reflecting conduction failure in a large fraction of SPNs. Losses were more pronounced in interganglionic pathways, suggesting increased vulnerability in branching projections. To assess activity-dependent effects, stimuli were delivered at 1, 5, and 20 Hz with focus on 5 and 20 Hz stimulus trains (20s duration). The overall time-course of train-evoked depression was similar across temperatures; however, the underlying axonal populations differed. At 22{degrees}C, slower-conducting axons exhibited marked frequency-dependent depression, whereas at 36{degrees}C the remaining faster-conducting axons displayed facilitation, particularly at 20 Hz. Slower-conducting responses also showed post-train potentiation at physiological temperature. These findings indicate that SPN axonal conduction is not uniformly reliable and is strongly modulated by temperature and activation history. Preferential vulnerability of slow-conducting, likely small-diameter and branching axons identifies axonal conduction as a physiologically regulated site of gain control in sympathetic output.

Prenatal alcohol exposure (PAE) causes fetal alcohol spectrum disorders (FASDs), which are neurodevelopmental conditions characterized by behavioral dysregulation, learning deficits, and cognitive inflexibilities. Alcohol exposure is harmful at all stages of human gestation, including the third trimester. This developmental window--characterized by rapid brain growth, myelination, and neural circuit formation--may be particularly vulnerable, yet the long-lasting behavioral and sensory consequences of exposure during this period remain poorly understood. In this study, neonatal mouse pups were exposed to ethanol (EtOH) or air vapor from postnatal day (P) 4 to P8, which is equivalent to a third-trimester alcohol exposure (TTAE) in humans. Blood ethanol concentrations measured at P8 reached approximately 250 mg/dL, consistent with binge-level exposure. Air- and EtOH-exposed mice were then assessed as adults at 5-6 months of age for locomotor activity, anxiety-related risky behaviors, recognition memory, and increased susceptibility to peripheral neuropathy, as indicated by sensitization to light touch following minor chronic constriction injury (mCCI) of the sciatic nerve. We found that TTAE was sufficient to produce long-lasting behavioral outcomes in a sex-dependent manner. Notably, EtOH-exposed males exhibited increased spontaneous locomotor activity and risky behavior, whereas EtOH-exposed females showed minimal or decreased changes compared to their respective controls. However, both EtOH-exposed male and female mice exhibited marked increases in light-touch sensitization, referred to as mechanical allodynia, following mCCI, a response absent in air-exposed controls. Together, these findings reveal that TTAE is highly detrimental to behavioral regulation and creates a vulnerability to developing neuropathic pain in adulthood.

Oligodendrocyte precursor cells (OPCs) are unique glial cells that communicate bidirectionally with neurons. Neuronal inputs drive various OPC behaviors, including proliferation and differentiation, immunomodulation, blood brain barrier regulation, synapse engulfment and axonal remodeling. OPCs are implicated in numerous stress and pain conditions, where their involvement is likely driven by neuronal activity (ie. neurotransmitter and neuropeptide signaling). One neuropeptide causally involved in chronic pain and stress conditions is calcitonin gene-related peptide (CGRP). Here, we tested the hypothesis that OPCs receive direct inputs from CGRP-containing neurons in the adult brain. Using RNAscope, immunofluorescence and analysis of single-cell datasets, we find that OPCs express receptors for CGRP and we identify close spatial contacts between CGRP and OPCs, with nearly half of CGRP puncta occurring within 1 {micro}m of an OPC. Some of these contacts appear to be synaptic, with CGRP-OPC contacts colocalizing with the presynaptic protein Bassoon and the postsynaptic protein PSD-95. This work suggests the presence of both diffuse and more direct forms of CGRP signaling to OPCs, raising the importance of future experiments to identify both the mode of CGRP release onto OPCs and the functional effects of these different contact types.

BackgroundTourette Syndrome (TS) is a neurodevelopmental movement disorder involving involuntary motor and vocal tics believed to be characterised by disordered neural inhibition. Cortical representations have previously been manipulated by disruptions in the inhibitory neurotransmitter {gamma}-aminobutyric acid (GABA). However, while facial tics are the most reported motor tic, it is unclear if facial sensorimotor representations differ in TS. MethodsSixteen individuals with Tourette Syndrome (TS) or chronic tic disorder and twenty typically developing (TD) control participants underwent 3-Tesla functional magnetic resonance imaging (fMRI). Blood-oxygenation level-dependent (BOLD) responses were measured during a block-design task comprising cued facial movements of common facial tics (blinking, grimacing and jaw clenching). Activations in bilateral pre- and post-central cortices and supplementary motor areas (SMA) were examined. Conjunction analyses identified voxels commonly and uniquely activated across movements within each group. ResultsBoth groups showed significant activations in the bilateral sensorimotor cortices and SMA in response to blink, grimace and jaw clench movements, with no significant between-group differences. Between-group similarities were lowest for unique blink maps. Common voxel maps also revealed low between-group similarity, with reduced sensorimotor activation and no shared SMA activation across movements in the TS group. ConclusionVoluntary facial sensorimotor representations do not differ between groups. However, low similarities between group unique blink maps may reflect greater prevalence of blinking tics in TS. Additionally, reduced overlap in sensorimotor activation and absent common SMA engagement across cued movements in the TS group may indicate altered motor integration or action initiation.

Alternative polyadenylation (APA) is a common posttranscriptional mechanism to regulate gene expression. APA generates mRNAs with varying lengths of 3' UTRs or transcripts that encode distinct protein carboxy-terminal ends. APA is especially important in neurons, where different mRNA variants are often asymmetrically localized to dendrites and axons, and can be locally translated into proteins. Local protein synthesis is crucial for axon guidance, synaptic plasticity, and learning and memory, key processes associated with the development of alcohol use disorder (AUD). We investigated the role of APA in AUD using a mouse model of alcohol dependence characterized by increased voluntary drinking after chronic intermittent ethanol (CIE) exposure. We examined APA during protracted withdrawal from alcohol in three brain regions of male and female mice. Our analyses revealed hundreds of genes undergoing APA in males, but substantially fewer in females, suggesting sex-specific effects of CIE on APA. Notably, male and female mice displayed distinct APA signatures. APA genes were different from differentially expressed genes (DEGs), suggesting that these molecular processes are regulated independently. We also determined that the expression of APA genes was associated with neurons, while DEGs were associated with non-neuronal cells. Many of the APA genes were involved in synaptic integrity, neuroplasticity, and neuronal maintenance, which was consistent with their enrichment in neurons. Our study suggests that APA is a crucial sex- and cell type-specific mechanism in AUD with the potential to influence localized neuronal protein expression during protracted withdrawal and to modify alcohol consumption behavior. HIGHLIGHTSO_LIChronic ethanol exposure in mice results in profound changes of APA genes in brain. C_LIO_LICommonly regulated cleavage and polyadenylation sites and genes were identified in male but not in female mice. C_LIO_LIThere was a minimal overlap between APA and differentially expressed genes (DEGs). C_LIO_LIAPA genes were primarily associated with neurons, whereas DEGs were associated with non-neuronal cells. C_LI