Biology subjects

Gross, C. T.

Publications and source records attributed to Gross, C. T..

17 recordsLinked to original sources

bioRxiv2026

Reciprocal feedback inhibition and divergent intrinsic firing properties between behaviorally defined classes of neurons in the mouse ventromedial hypothalamus

The capacity to orchestrate appropriate defensive behaviors in response to threatening stimuli is essential for the survival of organisms. Flight, freezing, appeasement, and attack, for example, are common defensive responses evoked depending upon the nature of the threat, the immediate context, and past experiences. Multiple brain regions have been implicated in regulating such responses, with the medial hypothalamus playing a key role in mediating innate responses to social and predator threats. The ventrolateral subdivision of the ventromedial hypothalamus (VMHvl) has been shown to mediate both avoidance and aggression towards conspecific threats in mice, suggesting that it serves as a circuit for context-appropriate defensive responses to social threats. Previous in vivo calcium imaging in VMHvl identified cells whose activity encodes either approach toward or escape from a social threat (Assessment+ and Flight+ cells, respectively), but it remained unclear how the switch in neural encoding from approach to avoidance occurs. Here, we use in vivo single-unit electrophysiology recordings coupled to channelrhodopsin-assisted circuit mapping (optrodes) to explore the functional and structural connectivity basis of the approach-avoidance switch. We confirm the presence of Assessment+ and Flight+ neurons in VMHvl and demonstrate that they are interconnected through feedback excitation and reciprocal feedback inhibition. Moreover, we discover that Assessment+ and Flight+ neurons exhibit a pronounced difference in their intrinsic firing properties. We hypothesize that this asymmetry in intrinsic responsivity coupled to reciprocal feedback inhibition underlies the nonlinear firing changes at the approach-to-avoidance transition and plays a role in triggering escape behavior.

neuroscience↗

bioRxiv2026

Fleeing is Believing: Adaptive behavior under social threat as an inference process

1.Adverse social experiences profoundly alter animal behavior, yet the processes underlying context-appropriate behavior selection based on prior social interactions remain poorly understood. Existing models capture statistical patterns but are not framed in mechanistic frameworks that explain individual variability and predict behavioral outcomes. We address this gap by modeling social defeat in mice as a partially observable Markov decision process (POMDP), implementing a heterarchical agent architecture - a structured network of interacting modules balancing exploration and exploitation. Our model successfully reconstructs observed behavioral motifs (e.g., investigation, hesitation, and flights), fits different mouse phenotypes (e.g., susceptible vs. resilient), and mechanistically captures the impact of social defeat as a parameter shift in the animals internal generative model. The model reproduces effects of interventions like optogenetic stimulation, and generates testable predictions for future experiments. The models modular architecture enables natural extension to other behavioral domains including foraging and multi-agent interactions, representing a foundational step toward interpretable models of mouse behavior. By capturing how adverse social experiences reshape decision-making at the computational level, this work offers potential clinical relevance for trauma and anxiety disorders in humans.

animal behavior and cognition↗

bioRxiv2026

Excitatory and inhibitory neurons in the dorsal periaqueductal gray encode decisions to assess and escape natural threats

Prey species are able to engage hardwired neural pathways to rapidly escape from an imminent predator attack. However, when predator threat is less probable they typically show a stereotypical sequence of approach toward the threat aimed at gathering more information, followed by escape to safety when the threat threshold is reached. The brainstem dorsal periaqueductal gray (dPAG) is required for the expression of escape behavior to predator threats and stimulation of dPAG elicits goal-directed flight. However, in vivo neural recordings in dPAG have identified separate populations of neurons that are tuned to either the approach or escape phase of the behavior suggesting that the structure may also be involved in threat assessment. The genetic identity and connectivity of these Assessment+ and Escape+ neurons have not been defined, although optogenetic activation of glutamatergic, but not GABAergic neurons elicits high-speed flight, suggesting that Escape+ neurons might be exclusively excitatory in nature. Moreover, it is not clear whether non-predator threats such as those elicited by conspecific or other animate threats are encoded by independent or overlapping neurons in dPAG. Here we report the activity pattern of ensembles of glutamatergic and GABAergic dPAG neurons during approach and escape from predator, social, and prey threats. Unexpectedly, we found that both glutamatergic and GABAergic neurons harbor Assessment+ and Escape+ neurons, suggesting that both cell-types are engaged in the approach-to-avoidance transition. Consistent with the functional involvement of both cell-types in approach-to-avoidance behavior, optogenetic activation of GABAergic cells elicited a reduction of risk assessment behavior towards the predator. Finally, we found that exposure to predator, social or prey threat recruited largely overlapping neurons in dPAG, demonstrating a convergence of threat processing in this structure. These findings point to a tightly coordinated role for dPAG excitatory and inhibitory neurons in the generalized control of innate threat assessment and avoidance behavior.

neuroscience↗

bioRxiv2025

Immediate early genes act in the medial hypothalamus to promote adaptation to social defeat

Territorial animals must moderate their social aggression and avoidance behaviors in a manner that maximizes their access to resources and fitness. The ventrolateral division of the ventromedial hypothalamic nucleus (VMHvl) has been shown to control both social aggression and avoidance in mice, and emerging data show that neural plasticity within VMHvl can drive the experience-dependent adaptation of these behaviors. Here, we investigated the contribution of immediate early gene (IEG) function in supporting this plasticity. In initial experiments, we found that downregulation of the IEG cFos in VMHvl did not significantly moderate the long-term increase in social avoidance seen following an experience of social defeat. However, local knockout of the IEG master regulator Serum Response Factor (SRF, Srf) was able to blunt the impact of social defeat on social avoidance and prevented the effect of social defeat on local optogenetic-evoked social avoidance behavior, demonstrating a critical role for subcortical IEG activity in social defeat-induced behavioral plasticity. To test whether NMDA receptor dependent plasticity might be involved in this effect, we locally infused the NMDA receptor antagonist MK-801 into VMHvl and assessed the impact of social defeat. Unexpectedly, MK-801 treatment led to an increase in social defeat-induced avoidance, pointing to the existence of opposing IEG and NMDA receptor-dependent adaptive mechanisms in medial hypothalamus. These findings suggest that multiple neural plasticity mechanisms are likely to be at play in the hypothalamic nuclei supporting innate behavior adaptation and show how IEG blockade can be used as a genetic tool to systematically map neural plasticity in the brain. Significance StatementThe role of IEGs in experience-dependent plasticity has not been investigated in subcortical structures until now. Our findings suggest IEGs expression is essential for adaptive behavioral changes in the VMHvl following social defeat and can potentially be used to map behavioral adaptation across brain regions. Our study also indicates a potential use of IEGs as targets of gene therapy for mitigating maladaptive behavioral adaptations.

neuroscience↗

bioRxiv2024

Induction of territorial behavior and dominance hierarchies in laboratory mice

Territorial behaviors comprise a set of coordinated actions and response patterns found across animal species that promote the exclusive access to resources. House mice are highly territorial with a subset of males consistently attacking and chasing competing males to expel them from their territories and performing urine marking behaviors to signal the extent of their territories. Natural variation in territorial behaviors within a mouse colony leads to the formation of dominance hierarchies in which subordinate males can reside within the territory of a dominant male. While the full repertoire of such territorial behaviors and hierarchies has been extensively studied in wild-derived mice in semi-natural enclosures, so far they have not been established in the smaller enclosures and with the genetically-defined laboratory strains required for the application of neural recording and manipulation methods. Here, we present a protocol to induce an extensive repertoire of territorial behaviors in small enclosures in laboratory mice, including a method for the simultaneous tracking of urine marking behavior in mouse pairs. Using this protocol we describe the emergence of robust dominant-subordinate hierarchies between pairs of CD1 outbred or CD1xB6 F1 hybrid mice, but unexpectedly not in C57BL/6 inbred animals. Our behavioral paradigm opens the door for neurocircuit studies of territorial behaviors and social hierarchy in the laboratory.

animal behavior and cognition↗

bioRxiv2021

Induction of flight via midbrain projections to thecuneiform nucleus

The dorsal periaqueductal gray is a midbrain structure implicated in the control of defensive behaviors and the processing of painful stimuli. Electrical stimulation or optogenetic activation of excitatory neurons in dorsal periaqueductal gray results in freezing or flight behavior at low or high intensity, respectively. However, the output structures that mediate these defensive behaviors remain unconfirmed. Here we carried out a targeted classification of neuron types in dorsal periaqueductal gray using multiplex in situ sequencing and then applied cell-type and projection-specific optogenetic stimulation to identify projections from dorsal periaqueductal grey to the cuneiform nucleus that promoted goal-directed flight behavior. These data confirmed that descending outputs from dorsal periaqueductal gray serve as a trigger for directed escape behavior.

neuroscience↗

bioRxiv2021

Identifying long-range synaptic inputs using genetically encoded labels and volume electron microscopy

Enzymes that facilitate the local deposition of electron dense reaction products have been widely used as labels in electron microscopy (EM). Peroxidases, in particular, can efficiently metabolize 3,3-diaminobenzidine tetrahydrochloride hydrate (DAB) to produce precipitates with high contrast under EM following heavy metal staining, and can be genetically encoded to facilitate the labeling of specific cell-types or organelles. Nevertheless, the peroxidase/DAB method has so far not been reported to work in combination with 3D volume EM techniques (e.g. Serial blockface electron microscopy, SBEM; Focused ion beam electron microscopy, FIBSEM) because the surfactant treatment needed for efficient reagent penetration disrupts tissue ultrastructure and because these methods require the deposition of large amounts of heavy metals that can obscure DAB precipitates. However, a recently described peroxidase with enhanced enzymatic activity (dAPEX2) appears to successfully deposit EM-visible DAB products in thick tissue without surfactant treatment. Here we demonstrate that multiplexed dAPEX2/DAB tagging is compatible with both FIBSEM and SBEM volume EM approaches and use them to map long-range genetically identified synaptic inputs from the anterior cingulate cortex to the periaqueductal gray in the mouse brain.

neuroscience↗

bioRxiv2021

Sensory-thresholded switch of neural firing states in a computational model of the ventromedial hypothalamus

The mouse ventromedial hypothalamus (VMH) is both necessary and sufficient for defensive responses to predator and social threats. Defensive behaviors typically involve cautious approach toward potentially threatening stimuli aimed at obtaining information about the risk involved, followed by sudden avoidance and flight behavior to escape harm. In vivo neural recording studies in mice have identified two major populations of VMH neurons that either increase their firing activity as the animal approaches the threat (called Assessment+ cells) or increase their activity as the animal flees the threat (called Flight+ cells). Interestingly, Assessment+ and Flight+ cells abruptly decrease and increase their firing activity, respectively, at the decision point for flight, creating an escape-related "switch" in functional state. This suggests that the activity of the two cell types in VMH is coordinated and could result from local circuit interactions. Here, we used computational modelling to test if a local inhibitory feedback circuit could give rise to key features of the neural activity seen in VMH during the approach-to-flight transition. Starting from a simple dual-population inhibitory feedback circuit receiving repeated trains of monotonically increasing sensory input to mimic approach to threat, we tested the requirement for balanced sensory input, balanced feedback, short-term synaptic plasticity, rebound excitation, and inhibitory feedback exclusivity to reproduce an abrupt, sensory-thresholded reciprocal firing change that resembles Assessment+ and Flight+ cell activity seen in vivo. Our work demonstrates that a relatively simple local circuit architecture is sufficient for the emergence of firing patterns similar to those seen in vivo and suggests that a reiterative process of experimental and computational work may be a fruitful avenue for better understanding the functional organization of mammalian instinctive behaviors at the circuit level.

neuroscience↗

bioRxiv2021

Microglia complement signaling promotes neuronal elimination and normal brain functional connectivity

Complement signaling is thought to serve as an opsonization signal to promote the phagocytosis of synapses by microglia. However, while its role in synaptic remodeling has been demonstrated in the retino-thalamic system, it remains unclear whether complement signaling mediates synaptic pruning in the brain more generally. Here we show that mice lacking the complement 3 receptor (C3r), the major microglia complement receptor, fail to show a deficit in either synaptic pruning or axon elimination in the developing mouse cortex. Instead, mice lacking C3r show a deficit in the perinatal elimination of neurons, both in the retina as well as in the cortex, a deficit that is associated with increased cortical thickness and enhanced functional connectivity in these regions in adulthood. These data demonstrate a preferential role for complement in promoting neuronal elimination in the developing brain and argue for a reconsideration of the role of complement in synaptic pruning.

neuroscience↗

bioRxiv2021

Microglia control glutamatergic synapses in the adult mouse hippocampus

Microglial cells are active players in regulating synaptic development and plasticity in the brain. However, how these cells influence the normal functioning of synapses is largely unknown. In this study, we characterized the effects of pharmacological depletion of microglia, achieved by administration of PLX5622, on hippocampal CA3-CA1 synapses of adult wild type mice. Following microglial depletion, we observed a reduction of spontaneous and evoked glutamatergic activity associated with a decrease of dendritic spine density. We also observed the appearance of immature synaptic features accompanied by higher levels of plasticity. In addition, microglia depleted mice showed a deficit in the acquisition of the Novel Object Recognition task. Remarkably, microglial repopulation after PLX5622 withdrawal was associated with the recovery of hippocampal synapses and learning functions. Altogether, these data demonstrate that microglia contribute to normal synaptic functioning in the adult brain and that their removal induces reversible changes in synaptic organization and activity of glutamatergic synapses.

neuroscience↗

bioRxiv2020

A role for cerebral cortex in the suppression of innate defensive behavior

The cerebral cortex is involved in the control of cognition and the processing of learned information and it appears to have a role in the adaptation of behavior in response to unpredictable circumstances. In addition, the cortex may have a role in the regulation of innate responses since rodents, cats or primates with surgical removal or accidental destruction of cortical regions show excessive irritability, aggression and rage elicited by threatening stimuli. However, it remains unclear whether cortex has an acute role in suppressing innate threat responses because the imprecision and chronic nature of these lesions leaves open the possibility that compensatory processes may underlie some of these phenotypes. In the present study we used pharmacogenetic inhibition to precisely, rapidly and reversibly suppress cortical pyramidal neuron function and examine its contribution to defensive behaviors elicited by a variety of innately aversive stimuli. Inhibition of cortex caused an increase of defensive responses elicited by an aggressive conspecific, a novel prey, and a physically stressful stimulus. These findings are consistent with a role of cortex in the acute inhibition of innate defensive behaviors.

neuroscience↗

bioRxiv2020

A maternal effect regulates global DNA methylation patterns

Variation in DNA methylation between individuals has been shown to be influenced by both genetic and environmental factors. However, the relative impact of genetic and non-genetic factors on DNA methylation patterns across the mammalian genome has not been systematically studied. We performed whole-genome methylation analysis in two inbred mouse strains, revealing striking differences in the global distribution of DNA methylation. Although global methylation patterns were indistinguishable for most genomic features, a significant increase in the number of unmethylated CpG-island promoters and first exons was observed between strains. Experiments using F1 reciprocal hybrid strains demonstrated that the genotype of the mother dictated global DNA methylation patterns. Cross-fostering experiments ruled out a postnatal maternal effect on these differences and suggested that they were driven by a prenatal maternal effect, possibly via differential deposition of maternal gene products into the oocyte or uterine environment. These data demonstrate that maternal effects have a major impact on global DNA methylation patterns.

genomics↗

bioRxiv2020

Mapping the neural circuitry of predator fear in the nonhuman primate

In rodents, innate and learned fear of predators depends on the medial hypothalamic defensive system, a conserved brain network that lies downstream of the amygdala. However, it remains unknown whether this network is involved in primate fear. Here we demonstrate that visually evoked predator fear recruits a homologous medial hypothalamic defense system in the nonhuman primate

neuroscience↗

bioRxiv2019

Dynamic encoding of social threat and spatial context in the hypothalamus

Territorial animals must be able to express social aggression or avoidance in a manner appropriate to spatial context and dominance status. Recent studies indicate that the ventromedial hypothalamus controls both innate aggression and avoidance, suggesting that it may encode an internal state of threat common to both behaviors. Here we used single unit in vivo calcium microendoscopy to identify neurons in the mouse ventromedial hypothalamus encoding social threat. Threat neurons were activated during social defeat as well as when the animal performed risk assessment. Unexpectedly, threat neurons were also activate in the chamber where the animal had been previously defeated and a distinct set of neurons emerged that were active in its home chamber, demonstrating the dynamic encoding of spatial context in the hypothalamus. Ensemble analysis of neural activity showed that social defeat induced a change in the encoding of social information and optogenetic activation of ventromedial hypothalamus neurons was able to elicit avoidance after, but not before social defeat, demonstrating a functional reorganization of the pathway by social experience. These findings reveal how instinctive behavior circuits in the hypothalamus dynamically encode spatial and sensory cues to drive adaptive social behaviors.

neuroscience↗

bioRxiv2018

Three-dimensional nanostructure of an intact microglia cell

Microglia are non-neuronal cells of the myeloid lineage that invade and take up long-term residence in the brain during development (Ginhoux et al. 2010) and are increasingly implicated in neuronal maturation, homeostasis, and pathology (Bessis et al. 2007; Paolicelli et al. 2011; Li et al. 2012; Aguzzi et al. 2013, Cunningham 2013, Cunningham et al. 2013). Since the early twentieth century several methods for staining and visualizing microglia have been developed. Scientists in Ramon y Cajals group (Achucarro 1913, Rio-Hortega 1919) pioneered these methods and their work led to the christening of microglia as the third element of the nervous system, distinct from astrocytes and neurons. More recently, a combination of imaging, genetic, and immunological tools has been used to visualize microglia in living brain (Davalos et al. 2005; Nimmerjahn et al. 2005). It was found that microglia are highly motile under resting conditions and rapidly respond to injuries (Kettenmann et al. 2011) suggesting a role for microglia in both brain homeostasis and pathology. Transmission Electron microscopy (TEM) has provided crucial complementary information on microglia morphology and physiology but until recently EM analyses have been limited to single or limited serial section studies (Tremblay et al. 2010; Paolicelli et al. 2011; Schafer et al. 2012; Tremblay et al. 2012; Sipe et al. 2016). TEM studies were successful in defining a set of morphological criteria for microglia: a polygonal nucleus with peripheral condensed chromatin, a relatively small cytoplasm with abundant presence of rough endoplasmic reticulum (RER), and a large volume of lysosomes and inclusions in the perikaryon. Recent advances in volumetric electron microscopy techniques allow for 3D reconstruction of large samples at nanometer-resolution, thus opening up new avenues for the understanding of cell biology and architecture in intact tissues. At the same time, correlative light and electron microscopy (CLEM) techniques have been extended to 3D brain samples to help navigate and identify critical molecular landmarks within large EM volumes (Briggman and Denk 2006; Maco et al. 2013; Blazquez-Llorca et al. 2015, Bosch et al. 2015). Here we present the first volumetric ultrastructural reconstruction of an entire mouse hippocampal microglia using serial block face scanning electron microscopy (SBEM). Using CLEM we have ensured the inclusion of both large, small, and filopodial microglia processes. Segmentation of the dataset allowed us to carry out a comprehensive inventory of microglia cell structures, including vesicles, organelles, membrane protrusions, and processes. This study provides a reference that can serve as a data mining resource for investigating microglia cell biology.

neuroscience↗

bioRxiv2018

Differential encoding of predator fear in the ventromedial hypothalamus and periaqueductal grey

The ventromedial hypothalamus is a central node of the mammalian predator defense network. Stimulation of this structure in rodents and primates elicits abrupt defensive responses, including flight, freezing, sympathetic activation, and panic, while inhibition reduces defensive responses to predators. The major efferent target of the ventromedial hypothalamus is the dorsal periaqueductal grey, and stimulation of this structure also elicits flight, freezing, and sympathetic activation. However, reversible inhibition experiments suggest that the ventromedial hypothalamus and periaqueductal grey play distinct roles in the control of defensive behavior, with the former proposed to encode an internal state necessary for the motivation of defensive responses, while the latter serves as a motor pattern initiator. Here we used electrophysiological recordings of single units in behaving mice exposed to a rat to investigate the encoding of predator fear in the dorsomedial division of the ventromedial hypothalamus and the dorsal periaqueductal grey. Distinct correlates of threat intensity and motor responses were found in both structures, suggesting a distributed encoding of sensory and motor features in the medial hypothalamic-brainstem instinctive network.

neuroscience↗

bioRxiv2017

Microglia remodel synapses by presynaptic trogocytosis and spine head filopodia induction

Microglia are highly motile glial cells that are proposed to mediate synaptic pruning during neuronal circuit formation. Disruption of signaling between microglia and neurons leads to an excess of immature synaptic connections, thought to be the result of impaired phagocytosis of synapses by microglia. However, until now the direct phagocytosis of synapses by microglia has not been reported and fundamental questions remain about the precise synaptic structures and phagocytic mechanisms involved. Here we used light sheet fluorescence microscopy to follow microglia-synapse interactions in developing organotypic hippocampal cultures, complemented by three-dimensional ultrastructural characterization using correlative light and electron microscopy (CLEM). Our findings define a set of dynamic microglia-synapse interactions, including the selective partial phagocytosis, or trogocytosis (trogo-: nibble), of presynaptic structures and the induction of postsynaptic spine head filopodia by microglia. These findings allow us to propose a mechanism for the facilitatory role of microglia in synaptic circuits remodeling and maturation.

neuroscience↗