Distinct neural mechanisms construct classical versus extraclassical inhibitory surrounds in an inhibitory nucleus in the midbrain attention network.

Inhibitory neurons in the midbrain spatial attention network, called isthmi pars magnocellularis (Imc), control stimulus selection by the sensorimotor and attentional hub, the optic tectum (OT). Here, we investigate in the barn owl how classical as well as extraclassical (global) inhibitory surrounds of Imc receptive fields (RFs), fundamental units of Imc computational function, are constructed. We find that focal, reversible blockade of GABAergic input onto Imc neurons disconnects their extraclassical inhibitory surrounds, but leaves intact their classical inhibitory surrounds. Subsequently, with paired recordings and iontophoresis, first at spatially aligned site-pairs in Imc and OT, and then, at mutually distant site-pairs within Imc, we demonstrate that classical inhibitory surrounds of Imc RFs are inherited from OT, but their extraclassical inhibitory surrounds are constructed within Imc. These results reveal key design principles of the midbrain spatial attention circuit and highlight the critical importance of competitive interactions within Imc for its operation.

Donut-like organization of inhibition underlies categorical neural responses in the midbrain.

Categorical neural responses underlie various forms of selection and decision-making. Such binary-like responses promote robust signaling of the winner in the presence of input ambiguity and neural noise. Here, we show that a 'donut-like' inhibitory mechanism in which each competing option suppresses all options except itself, is highly effective at generating categorical neural responses. It surpasses motifs of feedback inhibition, recurrent excitation, and divisive normalization invoked frequently in decision-making models. We demonstrate experimentally not only that this mechanism operates in the midbrain spatial selection network in barn owls, but also that it is necessary for categorical signaling by it. The functional pattern of neural inhibition in the midbrain forms an exquisitely structured 'multi-holed' donut consistent with this network's combinatorial inhibitory function for stimulus selection. Additionally, modeling reveals a generalizable neural implementation of the donut-like motif for categorical selection. Self-sparing inhibition may, therefore, be a powerful circuit module central to categorization.

Dynamics of Visual Perceptual Decision-Making in Freely Behaving Mice.

The temporal dynamics of perceptual decisions offer a key window into the cognitive processes contributing to decision-making. Investigating perceptual dynamics in a genetically tractable animal model can facilitate the subsequent unpacking of the underlying neural mechanisms. Here, we investigated the time course as well as fundamental psychophysical constants governing visual perceptual decision-making in freely behaving mice. We did so by analyzing response accuracy against reaction time (RT), i.e., conditional accuracy, in a series of two-alternative forced choice (2-AFC) orientation discrimination tasks in which we varied target size, luminance, duration, and presence of a foil. Our results quantified two distinct stages in the time course of mouse visual decision-making: a "sensory encoding" stage in which conditional accuracy exhibits a classic trade-off with response speed, and a subsequent "short-term memory (STM)-dependent" stage in which conditional accuracy exhibits a classic asymptotic decay following stimulus offset. We estimated the duration of visual sensory encoding as 200-320 ms across tasks, the lower bound of the duration of STM as ∼1700 ms, and the briefest duration of visual stimulus input that is informative as ≤50 ms. Separately, by varying stimulus onset delay, we demonstrated that the conditional accuracy function (CAF) and RT distribution can be independently modulated, and found that the duration for which mice naturally withhold from responding is a quantitative metric of impulsivity. Taken together, our results establish a quantitative foundation for investigating the neural circuit bases of visual decision dynamics in mice.

Neural Markers of Vulnerability to Anxiety Outcomes after Traumatic Brain Injury.

Anxiety outcomes after traumatic brain injury (TBI) are complex, and the underlying neural mechanisms are poorly understood. Here, we developed a multi-dimensional behavioral profiling approach to investigate anxiety-like outcomes in mice that takes into account individual variability. Departing from the tradition of comparing outcomes in TBI versus sham groups, we identified a subgroup within the TBI group that is vulnerable to anxiety dysfunction, and present increased exploration of the anxiogenic zone compared to sham controls or resilient injured animals, by applying dimensionality reduction, clustering, and post hoc validation to behavioral data obtained from multiple assays for anxiety at several post-injury time points. These vulnerable animals expressed distinct molecular profiles in the corticolimbic network, with downregulation in gamma-aminobutyric acid and glutamate and upregulation in neuropeptide Y markers. Indeed, among vulnerable animals, not resilient or sham controls, severity of anxiety-related outcomes correlated strongly with expression of molecular markers. Our results establish a foundational approach, with predictive power, for reliably identifying maladaptive anxiety outcomes after TBI and uncovering neural signatures of vulnerability to anxiety.

Mechanisms of competitive selection: A canonical neural circuit framework.

Competitive selection, the transformation of multiple competing sensory inputs and internal states into a unitary choice, is a fundamental component of animal behavior. Selection behaviors have been studied under several intersecting umbrellas including decision-making, action selection, perceptual categorization, and attentional selection. Neural correlates of these behaviors and computational models have been investigated extensively. However, specific, identifiable neural circuit mechanisms underlying the implementation of selection remain elusive. Here, we employ a first principles approach to map competitive selection explicitly onto neural circuit elements. We decompose selection into six computational primitives, identify demands that their execution places on neural circuit design, and propose a canonical neural circuit framework. The resulting framework has several links to neural literature, indicating its biological feasibility, and has several common elements with prominent computational models, suggesting its generality. We propose that this framework can help catalyze experimental discovery of the neural circuit underpinnings of competitive selection.

Endogenous and exogenous control of visuospatial selective attention in freely behaving mice.

Visuospatial selective attention has been investigated primarily in head-fixed animals and almost exclusively in primates. Here, we develop two human-inspired, discrimination-based behavioral paradigms for studying selective visuospatial attention in freely behaving mice. In the 'spatial probability' task, we find enhanced accuracy, sensitivity, and rate of evidence accumulation at the location with higher probability of target occurrence, and opposite effects at the lower probability location. Together with video-based 3D head-tracking, these results demonstrate endogenous expectation-driven shifts of spatial attention. In the 'flanker' task, we find that a second stimulus presented with the target, but with conflicting information, causes switch-like decrements in accuracy and sensitivity as a function of its contrast, and slower evidence accumulation, demonstrating exogenous capture of spatial attention. The ability to study primate-like selective attention rigorously in unrestrained mice opens a rich avenue for research into neural circuit mechanisms underlying this critical executive function in a naturalistic setting.

Categorical Signaling of the Strongest Stimulus by an Inhibitory Midbrain Nucleus.

The nucleus isthmi pars magnocellularis (Imc), a group of inhibitory neurons in the midbrain tegmentum, is a critical component of the spatial selection network in the vertebrate midbrain. It delivers long-range inhibition among different portions of the space map in the optic tectum (OT), thereby mediating stimulus competition in the OT. Here, we investigate the properties of relative strength-dependent competitive interactions within the Imc, in barn owls of both sexes. We find that when Imc neurons are presented simultaneously with one stimulus inside the receptive field and a second, competing stimulus outside, they exhibit gradual or switch-like response profiles as a function of relative stimulus strength. They do so both when the two stimuli are of the same sensory modality (both visual) or of different sensory modalities (visual and auditory). Moreover, Imc neurons signal the strongest stimulus in a dynamically flexible manner, indicating that Imc responses reflect an online comparison between the strengths of the competing stimuli. Notably, Imc neurons signal the strongest stimulus more categorically, and earlier than the OT. Paired recordings at spatially aligned sites in the Imc and OT reveal that although some properties of stimulus competition, such as the bias of competitive response profiles, are correlated, others such as the steepness of response profiles, are set independently. Our results demonstrate that the Imc is itself an active site of competition, and may be the first site in the midbrain selection network at which stimulus competition is resolved.SIGNIFICANCE STATEMENT This work sheds light on the functional properties of a small group of inhibitory neurons in the vertebrate midbrain that play a key part in how the brain selects a target among competitors. A better understanding of the functioning of these neurons is an important building block for the broader understanding of how distracters are suppressed, and of spatial attention and its dysfunction.

Distinct recruitment of dorsomedial and dorsolateral striatum erodes with extended training.

Hypotheses of striatal orchestration of behavior ascribe distinct functions to striatal subregions, with the dorsolateral striatum (DLS) especially implicated in habitual and skilled performance. Thus neural activity patterns recorded from the DLS, but not the dorsomedial striatum (DMS), should be correlated with habitual and automatized performance. Here, we recorded DMS and DLS neural activity in rats during training in a task promoting habitual lever pressing. Despite improving performance across sessions, clear changes in corresponding neural activity patterns were not evident in DMS or DLS during early training. Although DMS and DLS activity patterns were distinct during early training, their activity was similar following extended training. Finally, performance after extended training was not associated with DMS disengagement, as would be predicted from prior work. These results suggest that behavioral sequences may continue to engage both striatal regions long after initial acquisition, when skilled performance is consolidated.

Spatial Dependence of Stimulus Competition in the Avian Nucleus Isthmi Pars Magnocellularis.

The nucleus isthmi pars magnocellularis (Imc) is a group of specialized inhibitory neurons in the midbrain tegmentum, thought to be conserved across vertebrate classes. Past anatomical work in reptiles has suggested a role for it in stimulus selection, which has been supported by recent studies in avians. Additionally, focal inactivation of Imc neurons is known to abolish all competitive interactions in the optic tectum (OT; SC in mammals), a midbrain sensorimotor hub that is critical for the control of spatial attention, thereby revealing a key role for Imc in stimulus selection. However, the functional properties of Imc neurons are not well understood. Here, with electrophysiological experiments in the barn owl Imc, we show that Imc neurons themselves exhibit signatures of stimulus competition. Distant competing stimuli outside the spatial receptive field (RF) suppressed powerfully, and divisively, the responses of Imc neurons to stimuli inside the RF, and did so from all tested locations along the elevation as well as azimuth. Notably, this held true even for locations encoded by the opposite side of the brain from the one containing the recording site. This global divisive inhibition operated independently of the sensory modality of the competing stimulus. Thus, the Imc not only supplies inhibition to the OT to support competition there, but may itself be an active site of stimulus competition. These results from experiments in the barn owl shed light on the functional properties of a vital node in the vertebrate midbrain selection network.

Long-Term Effects of Traumatic Brain Injury on Anxiety-Like Behaviors in Mice: Behavioral and Neural Correlates.

Traumatic brain injury (TBI) has been frequently linked to affective disorders such as anxiety and depression. However, much remains to be understood about the underlying molecular and signaling mechanisms that mediate affective dysfunctions following injury. A lack of consensus in animal studies regarding what the affective sequelae of TBI are has been a major hurdle that has slowed progress, with studies reporting the full range of effects: increase, decrease, and no change in anxiety following injury. Here, we addressed this issue directly by investigating long-term anxiety outcomes in mice following a moderate to severe controlled cortical impact (CCI) injury using a battery of standard behavioral tests-the open field (OF), elevated zero maze (EZM), and elevated plus maze (EPM). Mice were tested on weeks 1, 3, 5 and 7 post-injury. Our results show that the effect of injury is time- and task-dependent. Early on-up to 3 weeks post-injury, there is an increase in anxiety-like behaviors in the elevated plus and zero mazes. However, after 5 weeks post-injury, anxiety-like behavior decreases, as measured in the OF and EZM. Immunostaining in the basolateral amygdala (BLA) for GAD, a marker for GABA, at the end of the behavioral testing showed the late decrease in anxiety behavior was correlated with upregulation of inhibition. The approach adopted in this study reveals a complex trajectory of affective outcomes following injury, and highlights the importance of comparing outcomes in different assays and time-points, to ensure that the affective consequences of injury are adequately assessed.

Shank and Zinc Mediate an AMPA Receptor Subunit Switch in Developing Neurons.

During development, pyramidal neurons undergo dynamic regulation of AMPA receptor (AMPAR) subunit composition and density to help drive synaptic plasticity and maturation. These normal developmental changes in AMPARs are particularly vulnerable to risk factors for Autism Spectrum Disorders (ASDs), which include loss or mutations of synaptic proteins and environmental insults, such as dietary zinc deficiency. Here, we show how Shank2 and Shank3 mediate a zinc-dependent regulation of AMPAR function and subunit switch from GluA2-lacking to GluA2-containing AMPARs. Over development, we found a concomitant increase in Shank2 and Shank3 with GluA2 at synapses, implicating these molecules as potential players in AMPAR maturation. Since Shank activation and function require zinc, we next studied whether neuronal activity regulated postsynaptic zinc at glutamatergic synapses. Zinc was found to increase transiently and reversibly with neuronal depolarization at synapses, which could affect Shank and AMPAR localization and activity. Elevated zinc induced multiple functional changes in AMPAR, indicative of a subunit switch. Specifically, zinc lengthened the decay time of AMPAR-mediated synaptic currents and reduced their inward rectification in young hippocampal neurons. Mechanistically, both Shank2 and Shank3 were necessary for the zinc-sensitive enhancement of AMPAR-mediated synaptic transmission and act in concert to promote removal of GluA1 while enhancing recruitment of GluA2 at pre-existing Shank puncta. These findings highlight a cooperative local dynamic regulation of AMPAR subunit switch controlled by zinc signaling through Shank2 and Shank3 to shape the biophysical properties of developing glutamatergic synapses. Given the zinc sensitivity of young neurons and its dependence on Shank2 and Shank3, genetic mutations and/or environmental insults during early development could impair synaptic maturation and circuit formation that underlie ASD etiology.

Combinatorial Neural Inhibition for Stimulus Selection across Space.

The ability to select the most salient among competing stimuli is essential for animal behavior and operates no matter which spatial locations stimuli happen to occupy. We provide evidence that the brain employs a combinatorially optimized inhibition strategy for selection across all pairs of stimulus locations. With experiments in a key inhibitory nucleus in the vertebrate midbrain selection network, called isthmi pars magnocellularis (Imc) in owls, we discovered that Imc neurons encode visual space with receptive fields that have multiple excitatory hot spots ("lobes"). Such multilobed encoding is necessitated by scarcity of Imc neurons. Although distributed seemingly randomly, the locations of these lobes are optimized across the high-firing Imc neurons, allowing them to combinatorially solve selection across space. This strategy minimizes metabolic and wiring costs, a principle that also accounts for observed asymmetries between azimuthal and elevational coding. Combinatorially optimized inhibition may be a general neural principle for efficient stimulus selection.

Mechanistic target of rapamycin is necessary for changes in dendritic spine morphology associated with long-term potentiation.

Alterations in the strength of excitatory synapses in the hippocampus is believed to serve a vital function in the storage and recall of new information in the mammalian brain. These alterations involve the regulation of both functional and morphological features of dendritic spines, the principal sites of excitatory synaptic contact. New protein synthesis has been implicated extensively in the functional changes observed following long-term potentiation (LTP), and changes to spine morphology have similarly been documented extensively following synaptic potentiation. However, mechanistic links between de novo translation and the structural changes of potentiated spines are less clear. Here, we assess explicitly the potential contribution of new protein translation under control of the mechanistic target of rapamycin (mTOR) to LTP-associated changes in spine morphology. Utilizing genetic and pharmacological manipulations of mTORC1 function in combination with confocal microscopy in live dissociated hippocampal cultures, we demonstrate that chemically-induced LTP (cLTP) requires do novo protein synthesis and intact mTORC1 signaling. We observed a striking diversity in response properties across morphological classes, with mushroom spines displaying a particular sensitivity to altered mTORC1 signaling across varied levels of synaptic activity. Notably, while pharmacological inhibition of mTORC1 signaling significantly diminished glycine-induced changes in spine morphology, transient genetic upregulation of mTORC1 signaling was insufficient to produce spine enlargements on its own. In contrast, genetic upregulation of mTORC1 signaling promoted rapid expansion in spine head diameter when combined with otherwise sub-threshold synaptic stimulation. These results suggest that synaptic activity-derived signaling pathways act in combination with mTORC1-dependent translational control mechanisms to ultimately regulate changes in spine morphology. As several monogenic neurodevelopmental disorders with links to Autism and Intellectual Disability share a common feature of dysregulated mTORC1 signaling, further understanding of the role of this signaling pathway in regulating synapse function and morphology will be essential in the development of novel therapeutic interventions.

Descending control of neural bias and selectivity in a spatial attention network: rules and mechanisms.

The brain integrates stimulus-driven (exogenous) activity with internally generated (endogenous) activity to compute the highest priority stimulus for gaze and attention. Little is known about how this computation is accomplished neurally. We explored the underlying functional logic in a critical component of the spatial attention network, the optic tectum (OT, superior colliculus in mammals), in awake barn owls. We found that space-specific endogenous influences, evoked by activating descending forebrain pathways, bias competition among exogenous influences, and substantially enhance the quality of the categorical neural pointer to the highest priority stimulus. These endogenous influences operate across sensory modalities. Biologically grounded modeling revealed that the observed effects on network bias and selectivity require a simple circuit mechanism: endogenously driven gain modulation of feedback inhibition among competing channels. Our findings reveal fundamental principles by which internal and external information combine to guide selection of the next target for gaze and attention.

Spatially reciprocal inhibition of inhibition within a stimulus selection network in the avian midbrain.

Reciprocal inhibition between inhibitory projection neurons has been proposed as the most efficient circuit motif to achieve the flexible selection of one stimulus among competing alternatives. However, whether such a motif exists in networks that mediate selection is unclear. Here, we study the connectivity within the nucleus isthmi pars magnocellularis (Imc), a GABAergic nucleus that mediates competitive selection in the midbrain stimulus selection network. Using laser photostimulation of caged glutamate, we find that feedback inhibitory connectivity is global within the Imc. Unlike typical lateral inhibition in other circuits, intra-Imc inhibition remains functionally powerful over long distances. Anatomically, we observed long-range axonal projections and retrograde somatic labeling from focal injections of bi-directional tracers in the Imc, consistent with spatial reciprocity of intra-Imc inhibition. Together, the data indicate that spatially reciprocal inhibition of inhibition occurs throughout the Imc. Thus, the midbrain selection circuit possesses the most efficient circuit motif possible for fast, reliable, and flexible selection.

A shared inhibitory circuit for both exogenous and endogenous control of stimulus selection.

The mechanisms by which the brain suppresses distracting stimuli to control the locus of attention are unknown. We found that focal, reversible inactivation of a single inhibitory circuit in the barn owl midbrain tegmentum, the nucleus isthmi pars magnocellularis (Imc), abolished both stimulus-driven (exogenous) and internally driven (endogenous) competitive interactions in the optic tectum (superior colliculus in mammals), which are vital to the selection of a target among distractors in behaving animals. Imc neurons transformed spatially precise multisensory and endogenous input into powerful inhibitory output that suppressed competing representations across the entire tectal space map. We identified a small, but highly potent, circuit that is employed by both exogenous and endogenous signals to exert competitive suppression in the midbrain selection network. Our findings reveal, to the best of our knowledge, for the first time, a neural mechanism for the construction of a priority map that is critical for the selection of the most important stimulus for gaze and attention.

Reciprocal inhibition of inhibition: a circuit motif for flexible categorization in stimulus selection.

As a precursor to the selection of a stimulus for gaze and attention, a midbrain network categorizes stimuli into "strongest" and "others." The categorization tracks flexibly, in real time, the absolute strength of the strongest stimulus. In this study, we take a first-principles approach to computations that are essential for such categorization. We demonstrate that classical feedforward lateral inhibition cannot produce flexible categorization. However, circuits in which the strength of lateral inhibition varies with the relative strength of competing stimuli categorize successfully. One particular implementation--reciprocal inhibition of feedforward lateral inhibition--is structurally the simplest, and it outperforms others in flexibly categorizing rapidly and reliably. Strong predictions of this anatomically supported circuit model are validated by neural responses measured in the owl midbrain. The results demonstrate the extraordinary power of a remarkably simple, neurally grounded circuit motif in producing flexible categorization, a computation fundamental to attention, perception, and decision making.

The role of a midbrain network in competitive stimulus selection.

A midbrain network interacts with the well-known frontoparietal forebrain network to select stimuli for gaze and spatial attention. The midbrain network, containing the superior colliculus (SC; optic tectum, OT, in non-mammalian vertebrates) and the isthmic nuclei, helps evaluate the relative priorities of competing stimuli and encodes them in a topographic map of space. Behavioral experiments in monkeys demonstrate an essential contribution of the SC to stimulus selection when the relative priorities of competing stimuli are similar. Neurophysiological results from the owl OT demonstrate a neural correlate of this essential contribution of the SC/OT. The multi-layered, spatiotopic organization of the midbrain network lends itself to the analysis and modeling of the mechanisms underlying stimulus selection for gaze and spatial attention.

Flexible categorization of relative stimulus strength by the optic tectum.

Categorization is the process by which the brain segregates continuously variable stimuli into discrete groups. We report that patterns of neural population activity in the owl optic tectum (OT) categorize stimuli based on their relative strengths into "strongest" versus "other." The category boundary shifts adaptively to track changes in the absolute strength of the strongest stimulus. This population-wide categorization is mediated by the responses of a small subset of neurons. Our data constitute the first direct demonstration of explicit categorization of stimuli by a neural network based on relative stimulus strength or salience. The finding of categorization by the population code relaxes constraints on the properties of downstream decoders that might read out the location of the strongest stimulus. These results indicate that the ensemble neural code in the OT could mediate bottom-up stimulus selection for gaze and attention, a form of stimulus categorization in which the category boundary often shifts within hundreds of milliseconds.

Signaling of the strongest stimulus in the owl optic tectum.

Essential to the selection of the next target for gaze or attention is the ability to compare the strengths of multiple competing stimuli (bottom-up information) and to signal the strongest one. Although the optic tectum (OT) has been causally implicated in stimulus selection, how it computes the strongest stimulus is unknown. Here, we demonstrate that OT neurons in the barn owl systematically encode the relative strengths of simultaneously occurring stimuli independently of sensory modality. Moreover, special "switch-like" responses of a subset of neurons abruptly increase when the stimulus inside their receptive field becomes the strongest one. Such responses are not predicted by responses to single stimuli and, indeed, are eliminated in the absence of competitive interactions. We demonstrate that this sensory transformation substantially boosts the representation of the strongest stimulus by creating a binary discrimination signal, thereby setting the stage for potential winner-take-all target selection for gaze and attention.