Adaptive robustness through incoherent signaling mechanisms in a regenerative brain.

Animal behavior emerges from collective dynamics of neurons, making it vulnerable to damage. Paradoxically, many organisms exhibit a remarkable ability to maintain significant behavior even after large-scale neural injury. Molecular underpinnings of this extreme robustness remain largely unknown. Here, we develop a quantitative pipeline to measure long-lasting latent states in planarian flatworm behaviors during whole-brain regeneration. By combining >20,000 animal trials with neural network modeling, we show that long-range volumetric peptidergic signals allow the planarian to rapidly restore coarse behavior output after large perturbations to the nervous system, while slow restoration of small-molecule neuromodulator functions refines precision. This relies on the different time and length scales of neuropeptide and small-molecule transmission to generate incoherent patterns of neural activity that competitively regulate behavior. Controlling behavior through opposing communication mechanisms creates a more robust system than either alone and may serve as a generalizable approach for constructing robust neural networks.

Feedforward and disinhibitory circuits differentially control activity of cortical somatostatin interneurons during behavioral state transitions.

Interneurons (INs), specifically those in disinhibitory circuits like somatostatin (SST) and vasoactive intestinal peptide (VIP)-INs, are strongly modulated by the behavioral context. Yet, the mechanisms by which these INs are recruited during active states and whether their activity is consistent across sensory cortices remain unclear. We now report that in mice, locomotor activity strongly recruits SST-INs in the primary somatosensory (S1) but not the visual (V1) cortex. This diverse engagement of SST-INs cannot be explained by differences in VIP-IN function but is absent in the presence of visual input, suggesting the involvement of feedforward sensory pathways. Accordingly, inactivating the somatosensory thalamus, but not decreasing VIP-IN activity, significantly reduces the modulation of SST-INs by locomotion. Model simulations suggest that the differences in SST-INs across behavioral states can be explained by varying ratios of VIP- and thalamus-driven activity. By integrating feedforward activity with neuromodulation, SST-INs are anticipated to be crucial for adapting sensory processing to behavioral states.

Automated behavioral analysis reveals that mice employ a bait-and-switch escape mechanism to de-escalate social conflict.

Intraspecies aggression has profound ecological and evolutionary consequences, as recipients can suffer injuries, decreases in fitness, and become outcasts from social groups. Although animals implement diverse strategies to avoid hostile confrontations, the extent to which social influences affect escape tactics is unclear. Here, we used computational and machine-learning approaches to analyze complex behavioral interactions as mixed-sex groups of mice, Mus musculus, freely interacted. Mice displayed a rich repertoire of behaviors marked by changes in behavioral state, aggressive encounters, and mixed-sex interactions. A prominent behavioral sequence consistently occurred after aggressive encounters, where males in submissive states quickly approached and transiently interacted with females immediately before the aggressor engaged with the same female. The behavioral sequences were also associated with substantially fewer physical altercations. Furthermore, the male's behavioral state and the interacting partners could be predicted by distinct features of the behavioral sequence, such as kinematics and the latency to and duration of male-female interactions. More broadly, our work revealed an ethologically relevant escape strategy influenced by the presence of females that may serve as a mechanism for de-escalating social conflict and preventing consequential reductions in fitness.

Sensory neurons couple arousal and foraging decisions in Caenorhabditis elegans.

Foraging animals optimize feeding decisions by adjusting both common and rare behavioral patterns. Here, we characterize the relationship between an animal's arousal state and a rare decision to leave a patch of bacterial food. Using long-term tracking and behavioral state classification, we find that food leaving decisions in Caenorhabditis elegans are coupled to arousal states across multiple timescales. Leaving emerges probabilistically over minutes from the high arousal roaming state, but is suppressed during the low arousal dwelling state. Immediately before leaving, animals have a brief acceleration in speed that appears as a characteristic signature of this behavioral motif. Neuromodulatory mutants and optogenetic manipulations that increase roaming have a coupled increase in leaving rates, and similarly acute manipulations that inhibit feeding induce both roaming and leaving. By contrast, inactivating a set of chemosensory neurons that depend on the cGMP-gated transduction channel TAX-4 uncouples roaming and leaving dynamics. In addition, tax-4-expressing sensory neurons promote lawn-leaving behaviors that are elicited by feeding inhibition. Our results indicate that sensory neurons responsive to both internal and external cues play an integrative role in arousal and foraging decisions.

When animals forage for food, they show distinct behavioral patterns in their movement. For instance, the nematode worm Caenorhabditis elegans shows two long-term behavioral states when exploring a patch of food: dwelling, when it moves slowly in a small area, and roaming, when it makes quick and wide-ranging movements. The worms will also occasionally suddenly decide to leave a piece of food and go explore the rest of their environment. Scientists know that the likelihood of the worms either roaming or dwelling is regulated by neurons passing molecules, such as serotonin and dopamine, to one another. However, it is not known how these two long-term behavioral states impact the momentary decision to leave a piece of food, and which mechanisms may regulate this coupling. To investigate, Scheer and Bargmann tracked the movement of genetically modified C. elegans and characterized their behavior. This revealed that the decision to leave food is not random but a distinct choice that primarily happens when worms are roaming. A characteristic signature of this response was that worms briefly accelerate immediately before leaving. Following this discovery, Scheer and Bargmann identified sensory neurons that are involved in this process. As well as detecting external sensory cues, these neurons also integrate internal signals, like whether the animal can eat, to specify how often a worm will leave food. The implications of this research extend beyond the realm of tiny nematodes. This study provides a new framework to examine the relationship between long-term behavior and momentary decision making. Such insights are crucial in understanding brain function across different organisms, including humans. It paves the way for further research into how behavior is regulated on multiple timescales in the brain.

Radial astrocyte synchronization modulates the visual system during behavioral-state transitions.

Glial cells support the function of neurons. Recent evidence shows that astrocytes are also involved in brain computations. To explore whether and how their excitable nature affects brain computations and motor behaviors, we used two-photon Ca2+ imaging of zebrafish larvae expressing GCaMP in both neurons and radial astrocytes (RAs). We found that in the optic tectum, RAs synchronize their Ca2+ transients immediately after the end of an escape behavior. Using optogenetics, ablations, and a genetically encoded norepinephrine sensor, we observed that RA synchronous Ca2+ events are mediated by the locus coeruleus (LC)-norepinephrine circuit. RA synchronization did not induce direct excitation or inhibition of tectal neurons. Nevertheless, it modulated the direction selectivity and the long-distance functional correlations among neurons. This mechanism supports freezing behavior following a switch to an alerted state. These results show that LC-mediated neuro-glial interactions modulate the visual system during transitions between behavioral states.

The Orienting Reflex Reveals Behavioral States Set by Demanding Contexts: Role of the Superior Colliculus.

Sensory stimuli can trigger an orienting reflex (response) by which animals move the head to position their sensors (e.g., eyes, pinna, whiskers). Orienting responses may be important to evaluate stimuli that call for action (e.g., approach, escape, ignore), but little is known about the dynamics of orienting responses in the context of goal-directed actions. Using mice of either sex, we found that, during a signaled avoidance action, the orienting response evoked by the conditioned stimulus (CS) consisted of a fast head movement containing rotational and translational components that varied substantially as a function of the behavioral and underlying brain states of the animal set by different task contingencies. Larger CS-evoked orienting responses were associated with high-intensity auditory stimuli, failures to produce the appropriate signaled action, and behavioral states resulting from uncertain or demanding situations and the animal's ability to cope with them. As a prototypical orienting neural circuit, we confirmed that the superior colliculus controls and codes the direction of spontaneous exploratory orienting movements. In addition, superior colliculus activity correlated with CS-evoked orienting responses, and either its optogenetic inhibition or excitation potentiated CS-evoked orienting responses, which are likely generated downstream in the medulla. CS-evoked orienting responses may be a useful probe to assess behavioral and related brain states, and state-dependent modulation of orienting responses may involve the superior colliculus.SIGNIFICANCE STATEMENT Humans and other animals produce an orienting reflex (also known as orienting response) by which they rapidly orient their head and sensors to evaluate novel or salient stimuli. Spontaneous orienting movements also occur during exploration of the environment in the absence of explicit, salient stimuli. We monitored stimulus-evoked orienting responses in mice performing signaled avoidance behaviors and found that these responses reflect the behavioral state of the animal set by contextual demands and the animal's ability to cope with them. Various experiments involving the superior colliculus revealed a well-established role in spontaneous orienting but only an influencing effect over orienting responses. Stimulus-evoked orienting responses may be a useful probe of behavioral and related brain states.

Diverse states and stimuli tune olfactory receptor expression levels to modulate food-seeking behavior.

Animals must weigh competing needs and states to generate adaptive behavioral responses to the environment. Sensorimotor circuits are thus tasked with integrating diverse external and internal cues relevant to these needs to generate context-appropriate behaviors. However, the mechanisms that underlie this integration are largely unknown. Here, we show that a wide range of states and stimuli converge upon a single Caenorhabditis elegans olfactory neuron to modulate food-seeking behavior. Using an unbiased ribotagging approach, we find that the expression of olfactory receptor genes in the AWA olfactory neuron is influenced by a wide array of states and stimuli, including feeding state, physiological stress, and recent sensory cues. We identify odorants that activate these state-dependent olfactory receptors and show that altered expression of these receptors influences food-seeking and foraging. Further, we dissect the molecular and neural circuit pathways through which external sensory information and internal nutritional state are integrated by AWA. This reveals a modular organization in which sensory and state-related signals arising from different cell types in the body converge on AWA and independently control chemoreceptor expression. The synthesis of these signals by AWA allows animals to generate sensorimotor responses that reflect the animal's overall state. Our findings suggest a general model in which sensory- and state-dependent transcriptional changes at the sensory periphery modulate animals' sensorimotor responses to meet their ongoing needs and states.

Spike-Wave Seizures, NREM Sleep and Micro-Arousals in WAG/Rij Rats with Genetic Predisposition to Absence Epilepsy: Developmental Aspects.

The current study was done in Wistar Albino Glaxo Rijswijk (WAG/Rij) rats, which are genetically prone to develop spontaneous spike-wave discharges (SWDs) and are widely used as a genetic model of absence epilepsy. Here, we examined functional links between sleep and spike-wave epilepsy in aging WAG/Rij rats using advanced techniques of EEG analysis. SWDs, periods of NREM sleep and micro-arousals were automatically detected in three-channel epidural EEG recorded in freely moving WAG/Rij rats consequently at the age 5, 7 and 9 months. We characterized the developmental profile of spike-wave epilepsy in drug-naïve WAG/Rij rats and defined three epi-phenotypes-severe, mild and minor epilepsy. Age-related changes of SWDs were associated with changes in NREM sleep. Several signs of NREM sleep fragmentation were defined in epileptic WAG/Rij rats. It seems that spike-wave epilepsy per se promotes micro-arousals during NREM sleep. However, subjects with a higher number of micro-arousals (and NREM sleep episodes) at the age of 5 months were characterized by a reduction of SWDs between 5 and 7 months of age.

Recording and Quantifying C. elegans Behavior.

Studies of C. elegans behavior have been crucial in identifying genetic pathways that control nervous system development and function, as well as basic principles of neural circuit function. Modern analysis of C. elegans behavior commonly relies on video recordings of animals, followed by automated image analysis and behavior quantification. Here, we describe two methods for recording and quantifying C. elegans behavior: a single-worm tracking approach that provides high-resolution behavioral data for individual animals and a multi-worm tracking approach that allows for quantification of the behavior of many animals in parallel. These approaches should be useful to a wide range of researchers studying the nervous system and behavior of C. elegans.

A behavioral approach to annotating sleep in infants: Building on the classic framework.

In infants, monitoring and assessment of sleep can offer valuable insights into sleep problems and neuro-cognitive development. The gold standard for sleep measurements is polysomnography (PSG), but this is rather obtrusive, and unpractical in non-laboratory situations. Behavioral observations constitute a non-obtrusive, infant-friendly alternative. In the current methodological paper, we describe and validate a behavior-based framework for annotating infant sleep states. For development of the framework, we used existing sleep data from an in-home study with an unobtrusive test setup. Participants were 20 infants with a mean age of 180 days. Framework development was based on Prechtl's method. We added rules and guidelines based on discussions and consent among annotators. Key to using our framework is combining data from several modalities, for example, closely observing the frequency, type, and quality of movements, breaths, and sounds an infant makes, while taking the context into account. For a first validation of the framework, we set up a small study with 14 infants (mean age 171 days), in which they took their day-time nap in a laboratory setting. They were continuously monitored by means of PSG, as well as by the test setup from the in-home study. Recordings were annotated based both on PSG and our framework, and then compared. Data showed that for scoring wake vs. active sleep vs. quiet sleep the framework yields results comparable to PSG with a Cohen's Kappa agreement of ≥0.74. Future work with a larger cohort is necessary for further validating this framework, and with clinical populations for determining whether it can be generalized to these populations as well.

Humpback whale adult females and calves balance acoustic contact with vocal crypsis during periods of increased separation.

Acoustic communication is important for animals with dependent young, particularly when they are spatially separated. Maternal humpback whales (Megaptera novaeangliae) use acoustic calling to help minimize the risk of separation from their young calves during migration. These pairs also use acoustic crypsis to minimize detection by males. How they balance a restricted active space with the need to maintain acoustic contact during periods of separation is not yet understood. Here, we analyzed movement metrics of tagged adult female-calf pairs during migration to identify two behavioral states, "resting/milling" and "travelling." When travelling, these pairs dived synchronously and exhibited little to no spatial separation. Alternatively, adult females had significantly longer dive durations (p < .01) when resting, and while they spent prolonged times at depth, calves would surface several times independently. This demonstrated that these pairs are frequently separated during periods of rest. We then determined whether the call rates and acoustic levels of these pairs increased with more frequent separation, finding that both adult females and calves significantly increased their call rates, but not levels, when resting. We also found that adult female-calf pairs have a restricted active space, with less than 15% of calls estimated to be detectable beyond 2 km. However, as with call level, detection distance did not differ significantly between the two behavioral states. In summary, adult female-calf pairs maintain successful communication during periods of separation by calling more frequently rather than by producing louder calls. This strategy aids in maintaining acoustic contact while simultaneously limiting detectability by conspecifics.

Absence of fetal heart rate cycling on the intrapartum cardiotocograph (CTG) is associated with intrapartum pyrexia and lower Apgar scores.

BACKGROUND: Cycling consists of alternating periods of reduced and normal fetal heart variability, reflecting changes in fetal behavioral states. Occurrence of active and quiet sleep cycles is considered to be a hallmark of fetal autonomic nervous system integrity, demonstrating healthy interaction between the parasympathetic and sympathetic nervous systems. Cycling is an overlooked feature in most international cardiotocography (CTG) guidelines. The authors tested the hypothesis that fetuses showing no cycling in the intrapartum period have poorer outcomes. AIM: To investigate whether the absence of cycling at the commencement of intrapartum fetal monitoring is associated with poorer neonatal outcomes (umbilical arterial cord pH, Apgar scores and neonatal unit admission). METHODS: Analysis of a database of sequentially acquired intrapartum CTG traces from a single center. Only cases of singleton pregnancies over 36 weeks gestation in cephalic presentation with recorded umbilical artery cord pH were considered. Neonatal outcomes were assessed based on umbilical cord artery pH, Apgar ≤7 at 5 min and unexpected admission to the neonatal unit. Intrapartum pyrexia, presence of meconium-stained amniotic fluid and mode of delivery were also recorded. RESULTS: A total of 684 cases were analyzed. Absence of cycling from the beginning of the intrapartum CTG recording was noted in 5% of cases. Cases with no cycling were more likely to have maternal pyrexia (≥37.8 °C) (p = .006) and Apgars ≤7 at 5 min (p = .04). There was an association between increasing baseline fetal heart rate and the proportion of cases with no cycling. There was no significant difference between the two groups with regard to the mode of delivery or umbilical cord arterial pH <7.05 (p = .53). CONCLUSION: Absence of cycling is associated with intrapartum maternal pyrexia and fetuses with the absence of cycling are more likely to have poorer perinatal outcomes measured by Apgar ≤ 7 at 5 min, despite no association with fetal acidosis. Results from this research were presented at the XXVI European Congress of Perinatal Medicine in September 2018.

Diving deeper into the underlying white shark behaviors at Guadalupe Island, Mexico.

Fine-scale movement patterns are driven by both biotic (hunting, physiological needs) and abiotic (environmental conditions) factors. The energy balance governs all movement-related strategic decisions.Marine environments can be better understood by considering the vertical component. From 24 acoustic trackings of 10 white sharks in Guadalupe Island, this study linked, for the first time, horizontal and vertical movement data and inferred six different behavioral states along with movement states, through the use of hidden Markov models, which allowed to draw a comprehensive picture of white shark behavior.Traveling was the most frequent state of behavior for white sharks, carried out mainly at night and twilight. In contrast, area-restricted searching was the least used, occurring primarily in daylight hours.Time of day, distance to shore, total shark length, and, to a lesser extent, tide phase affected behavioral states. Chumming activity reversed, in the short term and in a nonpermanent way, the behavioral pattern to a general diel vertical pattern.

Heart rate variability in neonatal seizures: Investigation and implications for management.

Many factors acting during the neonatal period can affect neurological development of the infant. Neonatal seizures (NS) that frequently occur in the immature brain may influence autonomic maturation and lead to detectable cardiovascular signs. These autonomic manifestations can also have significant diagnostic and prognostic value. The analysis of Heart Rate Variability (HRV) represents the most used and feasible method to evaluate cardiac autonomic regulation. This narrative review summarizes studies investigating HRV dynamics in newborns with seizures, with the aim of highlighting the potential utility of HRV measures for seizure detection and management. While HRV analysis in critically ill newborns is influenced by many potential confounders, we suggest that it can enhance the ability to better diagnose seizures in the clinical setting. We present potential applications of the analysis of HRV, which could have a useful future role, beyond the research setting.

A neural circuit for flexible control of persistent behavioral states.

To adapt to their environments, animals must generate behaviors that are closely aligned to a rapidly changing sensory world. However, behavioral states such as foraging or courtship typically persist over long time scales to ensure proper execution. It remains unclear how neural circuits generate persistent behavioral states while maintaining the flexibility to select among alternative states when the sensory context changes. Here, we elucidate the functional architecture of a neural circuit controlling the choice between roaming and dwelling states, which underlie exploration and exploitation during foraging in C. elegans. By imaging ensemble-level neural activity in freely moving animals, we identify stereotyped changes in circuit activity corresponding to each behavioral state. Combining circuit-wide imaging with genetic analysis, we find that mutual inhibition between two antagonistic neuromodulatory systems underlies the persistence and mutual exclusivity of the neural activity patterns observed in each state. Through machine learning analysis and circuit perturbations, we identify a sensory processing neuron that can transmit information about food odors to both the roaming and dwelling circuits and bias the animal towards different states in different sensory contexts, giving rise to context-appropriate state transitions. Our findings reveal a potentially general circuit architecture that enables flexible, sensory-driven control of persistent behavioral states.

Dorsal periaqueductal gray ensembles represent approach and avoidance states.

Animals must balance needs to approach threats for risk assessment and to avoid danger. The dorsal periaqueductal gray (dPAG) controls defensive behaviors, but it is unknown how it represents states associated with threat approach and avoidance. We identified a dPAG threatavoidance ensemble in mice that showed higher activity farther from threats such as the open arms of the elevated plus maze and a predator. These cells were also more active during threat avoidance behaviors such as escape and freezing, even though these behaviors have antagonistic motor output. Conversely, the threat approach ensemble was more active during risk assessment behaviors and near threats. Furthermore, unsupervised methods showed that avoidance/approach states were encoded with shared activity patterns across threats. Lastly, the relative number of cells in each ensemble predicted threat avoidance across mice. Thus, dPAG ensembles dynamically encode threat approach and avoidance states, providing a flexible mechanism to balance risk assessment and danger avoidance.

Home Team (Dis)Advantage Patterns in the National Hockey League: Changes Through Increased Emphasis on Individual Performance With the 3-on-3 Overtime Rule.

Past research examining National Hockey League (professional ice hockey; NHL) data from the 4-on-4 overtime era (seasons between 2005-06 and 2013-14) revealed an inconsistent home team (dis)advantage pattern such that home teams that were superior to their visiting counterparts had slightly greater odds of winning during regulation play compared to overtime (demonstrating home crowd advantages for team performance during regulation); in contrast, home teams experienced lower odds of winning in the shootout period than in overtime regardless of team quality (thereby demonstrating risks for individual choking from home crowd pressures). In this study, we explored the NHL home (dis)advantage pattern during four more recent seasons (2015-16 through 2018-19) in which the league instituted 3-on-3 play during overtime (perhaps increasing individual pressure for athletes competing in the 3-on-3 overtime period). We used archival data from the regular season (N=5,002 games) to compare home teams' odds of winning in regulation (with 5-on-5 skaters per team) to overtime (with 3-on-3) and in the shootout, adjusting for the quality of home and visiting teams. We conducted fixed-effects and multi-level logistic regression modeling. Evenly matched home teams were 1.66 times more likely to win than inferior home teams when games concluded in regulation versus overtime. Superior home teams were 4.24 times more likely to win than inferior home teams when games concluded in regulation rather than overtime. Thus, it is apparently more difficult for superior and evenly matched home teams to win in overtime than during regulation, suggesting that such home teams may be susceptible to choking in overtime. In contrast to the earlier 4-on-4 overtime era, home teams did not have lower odds of winning in the shootout compared to overtime. These results may have implications for NHL coaches' and players' tactical decision-making.

Phasic Activation of Dorsal Raphe Serotonergic Neurons Increases Pupil Size.

Transient variations in pupil size (PS) under constant luminance are coupled to rapid changes in arousal state,1-3 which have been interpreted as vigilance,4 salience,5 or a surprise signal.6-8 Neural control of such fluctuations presumably involves multiple brain regions5,9-11 and neuromodulatory systems,3,12,13 but it is often associated with phasic activity of the noradrenergic system.9,12,14,15 Serotonin (5-HT), a neuromodulator also implicated in aspects of arousal16 such as sleep-wake transitions,17 motivational state regulation,18 and signaling of unexpected events,19 seems to affect PS,20-24 but these effects have not been investigated in detail. Here we show that phasic 5-HT neuron stimulation causes transient PS changes. We used optogenetic activation of 5-HT neurons in the dorsal raphe nucleus (DRN) of head-fixed mice performing a foraging task. 5-HT-driven modulations of PS were maintained throughout the photostimulation period and sustained for a few seconds after the end of stimulation. We found no evidence that the increase in PS with activation of 5-HT neurons resulted from interactions of photostimulation with behavioral variables, such as locomotion or licking. Furthermore, we observed that the effect of 5-HT on PS depended on the level of environmental uncertainty, consistent with the idea that 5-HT could report a surprise signal.19 These results advance our understanding of the neuromodulatory control of PS, revealing a tight relationship between phasic activation of 5-HT neurons and changes in PS.

Mushroom Body Extrinsic Neurons in Walking Bumblebees Correlate With Behavioral States but Not With Spatial Parameters During Exploratory Behavior.

Central place foraging insects like honeybees and bumblebees learn to navigate efficiently between nest and feeding site. Essential components of this behavior can be moved to the laboratory. A major component of navigational learning is the active exploration of the test arena. These conditions have been used here to search for neural correlates of exploratory walking in the central arena (ground), and thigmotactic walking in the periphery (slope). We chose mushroom body extrinsic neurons (MBENs) because of their learning-related plasticity and their multi-modal sensitivities that may code relevant parameters in a brain state-dependent way. Our aim was to test whether MBENs code space-related components or are more involved in state-dependent processes characterizing exploration and thigmotaxis. MBENs did not respond selectively to body directions or locations. Their spiking activity differently correlated with walking speed depending on the animals' locations: on the ground, reflecting exploration, or on the slope, reflecting thigmotaxis. This effect depended on walking speed in different ways for different animals. We then asked whether these effects depended on spatial parameters or on the two states, exploration and thigmotaxis. Significant epochs of stable changes in spiking did not correlate with restricted locations in the arena, body direction, or walking transitions between ground and slope. We thus conclude that the walking speed dependencies are caused by the two states, exploration and thigmotaxis, rather than by spatial parameters.

Behavioral States.

Caenorhabditis elegans' behavioral states, like those of other animals, are shaped by its immediate environment, its past experiences, and by internal factors. We here review the literature on C. elegans behavioral states and their regulation. We discuss dwelling and roaming, local and global search, mate finding, sleep, and the interaction between internal metabolic states and behavior.