Neurophysiological and behavioral synchronization in group-living and sleeping mice.

Social interactions profoundly influence animal development, physiology, and behavior. Yet, how sleep-a central behavioral and neurophysiological process-is modulated by social interactions is poorly understood. Here, we characterized sleep behavior and neurophysiology in freely moving and co-living mice under different social conditions. We utilized wireless neurophysiological devices to simultaneously record multiple individuals within a group for 24 h, alongside video acquisition. We first demonstrated that mice seek physical contact before sleep initiation and sleep while in close proximity to each other (hereafter, "huddling"). To determine whether huddling during sleep is a motivated behavior, we devised a novel behavioral apparatus allowing mice to choose whether to sleep in close proximity to a conspecific or in solitude, under different environmental conditions. We also applied a deep-learning-based approach to classify huddling behavior. We demonstrate that mice are willing to forgo their preferred sleep location, even under thermoneutral conditions, to gain access to social contact during sleep. This strongly suggests that the motivation for prolonged physical contact-which we term somatolonging-drives huddling behavior. We then characterized sleep architecture under different social conditions and uncovered a social-dependent modulation of sleep. We also revealed coordination in multiple neurophysiological features among co-sleeping individuals, including in the timing of falling asleep and waking up and non-rapid eye movement sleep (NREMS) intensity. Notably, the timing of rapid eye movement sleep (REMS) was synchronized among co-sleeping male siblings but not co-sleeping female or unfamiliar mice. Our findings provide novel insights into the motivation for physical contact and the extent of social-dependent plasticity in sleep.

A Cortico-Striatal Circuit for Sound-Triggered Prediction of Reward Timing.

A crucial aspect of auditory perception is the ability to use sound cues to predict future events and to time actions accordingly. For example, distinct smartphone notification sounds reflect a call that needs to be answered within a few seconds, or a text that can be read later; the sound of an approaching vehicle signals when it is safe to cross the street. Other animals similarly use sounds to plan, time and execute behaviors such as hunting, evading predation and tending to offspring. However, the neural mechanisms that underlie sound-guided prediction of upcoming salient event timing are not well understood. To address this gap, we employed an appetitive sound-triggered reward time prediction behavior in head-fixed mice. We find that mice trained on this task reliably estimate the time from a sound cue to upcoming reward on the scale of a few seconds, as demonstrated by learning-dependent well-timed increases in reward-predictive licking. Moreover, mice showed a dramatic impairment in their ability to use sound to predict delayed reward when the auditory cortex was inactivated, demonstrating its causal involvement. To identify the neurophysiological signatures of auditory cortical reward-timing prediction, we recorded local field potentials during learning and performance of this behavior and found that the magnitude of auditory cortical responses to the sound prospectively encoded the duration of the anticipated sound-reward time interval. Next, we explored how and where these sound-triggered time interval prediction signals propagate from the auditory cortex to time and initiate consequent action. We targeted the monosynaptic projections from the auditory cortex to the posterior striatum and found that chemogenetic inactivation of these projections impairs animal's ability to predict sound-triggered delayed reward. Simultaneous neural recordings in the auditory cortex and posterior striatum during task performance revealed coordination of neural activity across these regions during the sound cue predicting the time interval to reward. Collectively, our findings identify an auditory cortical-striatal circuit supporting sound-triggered timing-prediction behaviors.

Exposure to sounds during sleep impairs hippocampal sharp wave ripples and memory consolidation.

Sleep is critical for the consolidation of recent experiences into long-term memories. As a key underlying neuronal mechanism, hippocampal sharp-wave ripples (SWRs) occurring during sleep define periods of hippocampal reactivation of recent experiences and have been causally linked with memory consolidation. Hippocampal SWR-dependent memory consolidation during sleep is often referred to as occurring during an "offline" state, dedicated to processing internally generated neural activity patterns rather than external stimuli. However, the brain is not fully disconnected from the environment during sleep. In particular, sounds heard during sleep are processed by a highly active auditory system which projects to brain regions in the medial temporal lobe, reflecting an anatomical pathway for sound modulation of hippocampal activity. While neural processing of salient sounds during sleep, such as those of a predator or an offspring, is evolutionarily adaptive, whether ongoing processing of environmental sounds during sleep interferes with SWR-dependent memory consolidation remains unknown. To address this question, we used a closed-loop system to deliver non-waking sound stimuli during or following SWRs in sleeping rats. We found that exposure to sounds during sleep suppressed the ripple power and reduced the rate of SWRs. Furthermore, sounds delivered during SWRs (On-SWR) suppressed ripple power significantly more than sounds delivered 2 seconds after SWRs (Off-SWR). Next, we tested the influence of sound presentation during sleep on memory consolidation. To this end, SWR-triggered sounds were applied during sleep sessions following learning of a conditioned place preference paradigm, in which rats learned a place-reward association. We found that On-SWR sound pairing during post-learning sleep induced a complete abolishment of memory retention 24 h following learning, while leaving memory retention immediately following sleep intact. In contrast, Off-SWR pairing weakened memory 24 h following learning as well as immediately following learning. Notably, On-SWR pairing induced a significantly larger impairment in memory 24 h after learning as compared to Off-SWR pairing. Together, these findings suggest that sounds heard during sleep suppress SWRs and memory consolidation, and that the magnitude of these effects are dependent on sound-SWR timing. These results suggest that exposure to environmental sounds during sleep may pose a risk for memory consolidation processes.

Auditory cortex ensembles jointly encode sound and locomotion speed to support sound perception during movement.

The ability to process and act upon incoming sounds during locomotion is critical for survival and adaptive behavior. Despite the established role that the auditory cortex (AC) plays in behavior- and context-dependent sound processing, previous studies have found that auditory cortical activity is on average suppressed during locomotion as compared to immobility. While suppression of auditory cortical responses to self-generated sounds results from corollary discharge, which weakens responses to predictable sounds, the functional role of weaker responses to unpredictable external sounds during locomotion remains unclear. In particular, whether suppression of external sound-evoked responses during locomotion reflects reduced involvement of the AC in sound processing or whether it results from masking by an alternative neural computation in this state remains unresolved. Here, we tested the hypothesis that rather than simple inhibition, reduced sound-evoked responses during locomotion reflect a tradeoff with the emergence of explicit and reliable coding of locomotion velocity. To test this hypothesis, we first used neural inactivation in behaving mice and found that the AC plays a critical role in sound-guided behavior during locomotion. To investigate the nature of this processing, we used two-photon calcium imaging of local excitatory auditory cortical neural populations in awake mice. We found that locomotion had diverse influences on activity of different neurons, with a net suppression of baseline-subtracted sound-evoked responses and neural stimulus detection, consistent with previous studies. Importantly, we found that the net inhibitory effect of locomotion on baseline-subtracted sound-evoked responses was strongly shaped by elevated ongoing activity that compressed the response dynamic range, and that rather than reflecting enhanced "noise," this ongoing activity reliably encoded the animal's locomotion speed. Decoding analyses revealed that locomotion speed and sound are robustly co-encoded by auditory cortical ensemble activity. Finally, we found consistent patterns of joint coding of sound and locomotion speed in electrophysiologically recorded activity in freely moving rats. Together, our data suggest that rather than being suppressed by locomotion, auditory cortical ensembles explicitly encode it alongside sound information to support sound perception during locomotion.

Enhanced stability of complex sound representations relative to simple sounds in the auditory cortex.

Typical everyday sounds, such as those of speech or running water, are spectrotemporally complex. The ability to recognize complex sounds (CxS) and their associated meaning is presumed to rely on their stable neural representations across time. The auditory cortex is critical for processing of CxS, yet little is known of the degree of stability of auditory cortical representations of CxS across days. Previous studies have shown that the auditory cortex represents CxS identity with a substantial degree of invariance to basic sound attributes such as frequency. We therefore hypothesized that auditory cortical representations of CxS are more stable across days than those of sounds that lack spectrotemporal structure such as pure tones (PTs). To test this hypothesis, we recorded responses of identified L2/3 auditory cortical excitatory neurons to both PTs and CxS across days using two-photon calcium imaging in awake mice. Auditory cortical neurons showed significant daily changes of responses to both types of sounds, yet responses to CxS exhibited significantly lower rates of daily change than those of PTs. Furthermore, daily changes in response profiles to PTs tended to be more stimulus-specific, reflecting changes in sound selectivity, as compared to changes of CxS responses. Lastly, the enhanced stability of responses to CxS was evident across longer time intervals as well. Together, these results suggest that spectrotemporally CxS are more stably represented in the auditory cortex across time than PTs. These findings support a role of the auditory cortex in representing CxS identity across time.Significance statementThe ability to recognize everyday complex sounds such as those of speech or running water is presumed to rely on their stable neural representations. Yet, little is known of the degree of stability of single-neuron sound responses across days. As the auditory cortex is critical for complex sound perception, we hypothesized that the auditory cortical representations of complex sounds are relatively stable across days. To test this, we recorded sound responses of identified auditory cortical neurons across days in awake mice. We found that auditory cortical responses to complex sounds are significantly more stable across days as compared to those of simple pure tones. These findings support a role of the auditory cortex in representing complex sound identity across time.

Lateral hypothalamic neuronal ensembles regulate pre-sleep nest-building behavior.

The transition from wakefulness to sleep requires striking alterations in brain activity, physiology, and behavior, yet the precise neuronal circuit elements facilitating this transition remain unclear. Prior to sleep onset, many animal species display characteristic behaviors, including finding a safe location, performing hygiene-related behaviors, and preparing a space for sleep. It has been proposed that the pre-sleep period is a transitional phase in which engaging in a specific behavioral repertoire de-arouses the brain and facilitates the wake-to-sleep transition, yet both causal evidence for this premise and an understanding of the neuronal circuit elements involved are lacking. Here, we combine detailed behavioral observations, EEG-EMG recordings, selective targeting, and activity modulation of pre-sleep-active neurons to reveal the behaviors preceding sleep initiation and their underlying neurobiological mechanisms. We show that mice engage in temporally structured behaviors with stereotypic EEG signatures prior to sleep and that nest-building and grooming become significantly more prevalent with sleep proximity. We next demonstrate that the ability to build a nest promotes the initiation and consolidation of sleep and that the lack of nesting material chronically fragments sleep. Lastly, we identify broadly projecting and predominantly glutamatergic neuronal ensembles in the lateral hypothalamus that regulate the motivation to engage in pre-sleep nest-building behavior and gate sleep initiation and intensity. Our study provides causal evidence for the facilitatory role of pre-sleep behaviors in sleep initiation and consolidation and a functional characterization of the neuronal underpinnings regulating a sleep-related and goal-directed complex behavior.

Encoding of acquired sound-sequence salience by auditory cortical offset responses.

Behaviorally relevant sounds are often composed of distinct acoustic units organized into specific temporal sequences. The meaning of such sound sequences can therefore be fully recognized only when they have terminated. However, the neural mechanisms underlying the perception of sound sequences remain unclear. Here, we use two-photon calcium imaging in the auditory cortex of behaving mice to test the hypothesis that neural responses to termination of sound sequences ("Off-responses") encode their acoustic history and behavioral salience. We find that auditory cortical Off-responses encode preceding sound sequences and that learning to associate a sound sequence with a reward induces enhancement of Off-responses relative to responses during the sound sequence ("On-responses"). Furthermore, learning enhances network-level discriminability of sound sequences by Off-responses. Last, learning-induced plasticity of Off-responses but not On-responses lasts to the next day. These findings identify auditory cortical Off-responses as a key neural signature of acquired sound-sequence salience.

Author Correction: Functional organization and population dynamics in the mouse primary auditory cortex.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Arousal State-Dependent Alterations in VTA-GABAergic Neuronal Activity.

Decades of research have implicated the ventral tegmental area (VTA) in motivation, learning and reward processing. We and others recently demonstrated that it also serves as an important node in sleep/wake regulation. Specifically, VTA-dopaminergic neuron activation is sufficient to drive wakefulness and necessary for the maintenance of wakefulness. However, the role of VTA-GABAergic neurons in arousal regulation is not fully understood. It is still unclear whether VTA-GABAergic neurons predictably alter their activity across arousal states, what is the nature of interactions between VTA-GABAergic activity and cortical oscillations, and how activity in VTA-GABAergic neurons relates to VTA-dopaminergic neurons in the context of sleep/wake regulation. To address these, we simultaneously recorded population activity from VTA subpopulations and electroencephalography/electromyography (EEG/EMG) signals during spontaneous sleep/wake states and in the presence of salient stimuli in freely-behaving mice. We found that VTA-GABAergic neurons exhibit robust arousal-state-dependent alterations in population activity, with high activity and transients during wakefulness and REM sleep. During wakefulness, population activity of VTA-GABAergic neurons, but not VTA-dopaminergic neurons, was positively correlated with EEG γ power and negatively correlated with θ power. During NREM sleep, population activity in both VTA-GABAergic and VTA-dopaminergic neurons negatively correlated with δ, θ, and σ power bands. Salient stimuli, with both positive and negative valence, activated VTA-GABAergic neurons. Together, our data indicate that VTA-GABAergic neurons, like their dopaminergic counterparts, drastically alter their activity across sleep-wake states. Changes in their activity predicts cortical oscillatory patterns reflected in the EEG, which are distinct from EEG spectra associated with dopaminergic neural activity.

The transformation of multi-sensory experiences into memories during sleep.

Our everyday lives present us with a continuous stream of multi-modal sensory inputs. While most of this information is soon forgotten, sensory information associated with salient experiences can leave long-lasting memories in our minds. Extensive human and animal research has established that the hippocampus is critically involved in this process of memory formation and consolidation. However, the underlying mechanistic details are still only partially understood. Specifically, the hippocampus has often been suggested to encode information during experience, temporarily store it, and gradually transfer this information to the cortex during sleep. In rodents, ample evidence has supported this notion in the context of spatial memory, yet whether this process adequately describes the consolidation of multi-sensory experiences into memories is unclear. Here, focusing on rodent studies, I examine how multi-sensory experiences are consolidated into long term memories by hippocampal and cortical circuits during sleep. I propose that in contrast to the classical model of memory consolidation, the cortex is a "fast learner" that has a rapid and instructive role in shaping hippocampal-dependent memory consolidation. The proposed model may offer mechanistic insight into memory biasing using sensory cues during sleep.

A cortical-hippocampal-cortical loop of information processing during memory consolidation.

Hippocampal replay during sharp-wave ripple events (SWRs) is thought to drive memory consolidation in hippocampal and cortical circuits. Changes in neocortical activity can precede SWR events, but whether and how these changes influence the content of replay remains unknown. Here we show that during sleep there is a rapid cortical-hippocampal-cortical loop of information flow around the times of SWRs. We recorded neural activity in auditory cortex (AC) and hippocampus of rats as they learned a sound-guided task and during sleep. We found that patterned activation in AC precedes and predicts the subsequent content of hippocampal activity during SWRs, while hippocampal patterns during SWRs predict subsequent AC activity. Delivering sounds during sleep biased AC activity patterns, and sound-biased AC patterns predicted subsequent hippocampal activity. These findings suggest that activation of specific cortical representations during sleep influences the identity of the memories that are consolidated into long-term stores.

VTA dopaminergic neurons regulate ethologically relevant sleep-wake behaviors.

Dopaminergic ventral tegmental area (VTA) neurons are critically involved in a variety of behaviors that rely on heightened arousal, but whether they directly and causally control the generation and maintenance of wakefulness is unknown. We recorded calcium activity using fiber photometry in freely behaving mice and found arousal-state-dependent alterations in VTA dopaminergic neurons. We used chemogenetic and optogenetic manipulations together with polysomnographic recordings to demonstrate that VTA dopaminergic neurons are necessary for arousal and that their inhibition suppresses wakefulness, even in the face of ethologically relevant salient stimuli. Nevertheless, before inducing sleep, inhibition of VTA dopaminergic neurons promoted goal-directed and sleep-related nesting behavior. Optogenetic stimulation, in contrast, initiated and maintained wakefulness and suppressed sleep and sleep-related nesting behavior. We further found that different projections of VTA dopaminergic neurons differentially modulate arousal. Collectively, our findings uncover a fundamental role for VTA dopaminergic circuitry in the maintenance of the awake state and ethologically relevant sleep-related behaviors.

Coordinated Excitation and Inhibition of Prefrontal Ensembles during Awake Hippocampal Sharp-Wave Ripple Events.

Interactions between the hippocampus and prefrontal cortex (PFC) are critical for learning and memory. Hippocampal activity during awake sharp-wave ripple (SWR) events is important for spatial learning, and hippocampal SWR activity often represents past or potential future experiences. Whether or how this reactivation engages the PFC, and how reactivation might interact with ongoing patterns of PFC activity, remains unclear. We recorded hippocampal CA1 and PFC activity in animals learning spatial tasks and found that many PFC cells showed spiking modulation during SWRs. Unlike in CA1, SWR-related activity in PFC comprised both excitation and inhibition of distinct populations. Within individual SWRs, excitation activated PFC cells with representations related to the concurrently reactivated hippocampal representation, while inhibition suppressed PFC cells with unrelated representations. Thus, awake SWRs mark times of strong coordination between hippocampus and PFC that reflects structured reactivation of representations related to ongoing experience.

Global order and local disorder in brain maps.

Maps serve as a ubiquitous organizing principle in the mammalian brain. In several sensory systems, such as audition, vision, and somatosensation, topographic maps are evident throughout multiple levels of brain pathways. Topographic maps, like retinotopy and tonotopy, persist from the receptor surface up to the cortex. Other maps, such as those of orientation preference in the visual cortex, are first created in the cortex itself. Despite the prevalence of topographic maps, it is still not clear what function they subserve. Although maps are topographically smooth at the macroscale, they are often locally heterogeneous. Here, we review studies describing the anatomy and physiology of topographic maps across various spatial scales, from the smooth macroscale to the heterogeneous local microarchitecture, with emphasis on maps of the visual and auditory systems. We discuss the potential advantages of local heterogeneity in brain maps, how they reflect complex cortical connectivity, and how they may impact sensory coding and local computations.

Elevated correlations in neuronal ensembles of mouse auditory cortex following parturition.

The auditory cortex is malleable by experience. Previous studies of auditory plasticity have described experience-dependent changes in response profiles of single neurons or changes in global tonotopic organization. However, experience-dependent changes in the dynamics of local neural populations have remained unexplored. In this study, we examined the influence of a dramatic yet natural experience in the life of female mice, giving birth and becoming a mother on single neurons and neuronal ensembles in the primary auditory cortex (A1). Using in vivo two-photon calcium imaging and electrophysiological recordings from layer 2/3 in A1 of mothers and age-matched virgin mice, we monitored changes in the responses to a set of artificial and natural sounds. Population dynamics underwent large changes as measured by pairwise and higher-order correlations, with noise correlations increasing as much as twofold in lactating mothers. Concomitantly, changes in response properties of single neurons were modest and selective. Remarkably, despite the large changes in correlations, information about stimulus identity remained essentially the same in the two groups. Our results demonstrate changes in the correlation structure of neuronal activity as a result of a natural life event.

Multisensory integration of natural odors and sounds in the auditory cortex.

VIDEO ABSTRACT: Motherhood is associated with different forms of physiological alterations including transient hormonal changes and brain plasticity. The underlying impact of these changes on the emergence of maternal behaviors and sensory processing within the mother's brain are largely unknown. By using in vivo cell-attached recordings in the primary auditory cortex of female mice, we discovered that exposure to pups' body odor reshapes neuronal responses to pure tones and natural auditory stimuli. This olfactory-auditory interaction appeared naturally in lactating mothers shortly after parturition and was long lasting. Naive virgins that had experience with the pups also showed an appearance of olfactory-auditory integration in A1, suggesting that multisensory integration may be experience dependent. Neurons from lactating mothers were more sensitive to sounds as compared to those from experienced mice, independent of the odor effects. These uni- and multisensory cortical changes may facilitate the detection and discrimination of pup distress calls and strengthen the bond between mothers and their neonates.

Functional organization and population dynamics in the mouse primary auditory cortex.

Cortical processing of auditory stimuli involves large populations of neurons with distinct individual response profiles. However, the functional organization and dynamics of local populations in the auditory cortex have remained largely unknown. Using in vivo two-photon calcium imaging, we examined the response profiles and network dynamics of layer 2/3 neurons in the primary auditory cortex (A1) of mice in response to pure tones. We found that local populations in A1 were highly heterogeneous in the large-scale tonotopic organization. Despite the spatial heterogeneity, the tendency of neurons to respond together (measured as noise correlation) was high on average. This functional organization and high levels of noise correlations are consistent with the existence of partially overlapping cortical subnetworks. Our findings may account for apparent discrepancies between ordered large-scale organization and local heterogeneity.