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Observational fear (OF) is the ability to vicariously experience and learn from another's fearful situation, enabling adaptive responses crucial for survival. It has been shown that the anterior cingulate cortex (ACC) and basolateral amygdala (BLA) are crucial for OF. A subset of neurons in the ACC is activated when observing aversive events in the demonstrator, which elicits OF. However, the neural circuit mechanisms underlying the expression of OF-related activity in the ACC remain unexplored. Previous studies have shown that the mediodorsal thalamus (MD) is crucial for OF, and MD neurons project to the ACC. Therefore, we hypothesize that the projection from MD to ACC may facilitate the OF-related activity in the ACC. By utilizing in vivo calcium imaging combined with the optogenetic terminal inhibition of MD-ACC pathway, we found that a subset of ACC neurons was activated when observing demonstrator's fearful situation in male mice. Furthermore, the optogenetic inhibition of the MD-ACC projection during the demonstrator's aversive moments significantly suppressed the OF-related activity in the ACC. Our data suggests that the MD-ACC projection plays a role in OF-related activity in ACC neurons. [ABSTRACT FROM AUTHOR]

The hippocampal dentate gyrus (DG) is thought to orthogonalize inputs from the entorhinal cortex (pattern separation) and relay this information to the CA3 region. In turn, attractor dynamics in CA3 perform a pattern completion or error correction operation before sending its output to CA1. In a mouse model of congenital hypoplasia of the DG, a deficiency in the Wntless (Wls) gene, specifically in cells expressing Gfap-Cre, which targets neuronal progenitors, led to an almost total absence of dentate granule cells and modestly impaired performance in spatial tasks. Here, we investigated the physiological consequences of granule cell loss in these mice by conducting in vivo calcium imaging from CA1 principal cells during behavior. The spatial selectivity of these cells was preserved without the DG. On a linear track, place fields in mutant mice were more likely to be near track terminals and to encode the distance from the start point in each running direction. In an open box, CA1 cells in mutant mice exhibited reductions in the percentage of place cells, in spatial information, and in place field stability. The reduction in place field stability across repeated exposures to the same environment resulted in a reduction in the differential representations of two different contexts in mutant mice compared to wild-type mice. These results suggest that DG helps to stabilize CA1 spatial representations, especially in 2-D environments, and that the lack of stability across similar environments may play a key role in the deficits of animals with DG dysfunction in discriminating different environments. [ABSTRACT FROM AUTHOR]

Neonatal hyperbilirubinemia may result in long-lasting motor, auditory and learning impairments. The mechanisms responsible for the localization of unconjugated bilirubin (UCB) to specific brain areas as well as those involved in potentially permanent central nervous system (CNS) dysfunctions are far from being clear. One area of investigation includes exploring how hyperbilirubinemia determines neuronal alterations predisposing to neurodevelopmental disorders. We focused on the hippocampus and pyramidal cell dysregulation of calcium homeostasis and synaptic activity, with a particular focus on early forms of correlated network activity, i.e., giant depolarizing potentials (GDPs), crucially involved in shaping mature synaptic networks. We performed live calcium imaging and patch clamp recordings from acute hippocampal slices isolated from wild-type rats exposed to exogenous high bilirubin concentration. We then explored the impact of endogenous bilirubin accumulation in hippocampal slices isolated from a genetic model of hyperbilirubinemia, i.e., Gunn rats. Our data show in both models an age-dependent dysregulation of calcium dynamics accompanied by severe alterations in GDPs, which were strongly reduced in hippocampal slices of hyperbilirubinemic rats, where the expression of GABAergic neurotransmission markers was also altered. We propose that hyperbilirubinemia damages neurons and affects the refinement of GABAergic synaptic circuitry during a critical period of hippocampal development. [ABSTRACT FROM AUTHOR]

Supplemental Digital Content is Available in the Text. This in vivo calcium imaging study shows that spinal cord stimulation inhibited the activation of superficial neurokinin 1 receptor–positive neurons, unraveling a new mechanism for spinal cord stimulation–induced analgesia. Spinal cord stimulation (SCS) is an effective modality for pain treatment, yet its underlying mechanisms remain elusive. Neurokinin 1 receptor–positive (NK1R+) neurons in spinal lamina I play a pivotal role in pain transmission. To enhance our mechanistic understanding of SCS-induced analgesia, we investigated how different SCS paradigms modulate the activation of NK1R+ neurons, by developing NK1R-Cre;GCaMP6s transgenic mice and using in vivo calcium imaging of superficial NK1R+ neurons under anesthesia (1.5% isoflurane). Neurokinin 1 receptor–positive neurons in the lumbar spinal cord (L4-5) showed a greater activation by electrical test stimulation (TS, 3.0 mA, 1 Hz) at the hindpaw at 2 weeks after tibia-sparing nerve injury (SNI-t) than in naïve mice. Spinal cord stimulation was then delivered through a bipolar plate electrode placed epidurally at L1-2 level. The short-term 50-Hz high-intensity SCS (80% motor threshold [MoT], 10 minutes) induced robust and prolonged inhibition of NK1R+ neuronal responses to TS in both naïve and SNI-t mice. The 30-minute 50-Hz and 900-Hz SCS applied at moderate intensity (50% MoT) also significantly inhibited neuronal responses in SNI-t mice. However, at low intensity (20% MoT), the 30-minute 900-Hz SCS only induced persistent neuronal inhibition in naïve mice, but not in SNI-t mice. In conclusion, both 10-minute high-intensity SCS and 30-minute SCS at moderate intensity inhibit the activation of superficial NK1R+ neurons, potentially attenuating spinal nociceptive transmission. Furthermore, in vivo calcium imaging of NK1R+ neurons provides a new approach for exploring the spinal neuronal mechanisms of pain inhibition by neuromodulation pain therapies. [ABSTRACT FROM AUTHOR]

The generation of retinal models from human induced pluripotent stem cells holds significant potential for advancing our understanding of retinal development, neurodegeneration, and the in vitro modeling of neurodegenerative disorders. The retina, as an accessible part of the central nervous system, offers a unique window into these processes, making it invaluable for both study and early diagnosis. This study investigates the impact of the Frontotemporal Dementia-linked IVS 10 + 16 MAPT mutation on retinal development and function using 2D and 3D retinal models derived from human induced pluripotent stem cells. Our findings reveal that the MAPT mutation leads to delayed retinal cell differentiation and maturation, with tau-mutant disease models exhibiting sustained higher expression of retinal progenitor cell markers and a reduced presence of post-mitotic neurons. Both 2D and 3D tau-mutant retinal models demonstrated an imbalance in tau isoforms, favoring 4R tau, along with increased tau phosphorylation, altered neurite morphology, and impaired cytoskeletal maturation. These changes are associated with impaired synaptic development, reduced neuronal connectivity, and enhanced cellular stress responses, including the increased formation of stress granules, markers of apoptosis and autophagy, and the presence of intracellular toxic tau aggregates. This study highlights the value of retinal models derived from human induced pluripotent stem cells in exploring the mechanisms underlying retinal pathology associated with tau mutations. These models offer essential insights into the development of therapeutic strategies for neurodegenerative diseases characterized by tau aggregation. [ABSTRACT FROM AUTHOR]

Human and non-human primate studies clearly implicate the dorsolateral prefrontal cortex (dlPFC) as critical for advanced cognitive functions. 1,2 It is thought that intracortical synaptic architectures within the dlPFC are the integral neurobiological substrate that gives rise to these processes. 3,4,5,6,7 In the prevailing model, each cortical column makes up one fundamental processing unit composed of dense intrinsic connectivity, conceptualized as the "canonical" cortical microcircuit. 3,8 Each cortical microcircuit receives sensory and cognitive information from upstream sources, which are represented by sustained activity within the microcircuit, referred to as persistent or recurrent activity. 4,9 Via recurrent connections within the microcircuit, activity propagates for a variable length of time, thereby allowing temporary storage and computations to occur locally before ultimately passing a transformed representation to a downstream output. 4,5,10 Competing theories regarding how microcircuit activity is coordinated have proven difficult to reconcile in vivo , where intercortical and intracortical computations cannot be fully dissociated. 5,9,11,12 Here, using high-density calcium imaging of macaque dlPFC, we isolated intracortical computations by interrogating microcircuit networks ex vivo. Using peri-sulcal stimulation to evoke recurrent activity in deep layers, we found that activity propagates through stochastically assembled intracortical networks wherein orderly, predictable, low-dimensional collective dynamics arise from ensembles with highly labile cellular memberships. Microcircuit excitability covaried with individual cognitive performance, thus anchoring heuristic models of abstract cortical functions within quantifiable constraints imposed by the underlying synaptic architecture. Our findings argue against engram or localist architectures, together demonstrating that generation of high-fidelity population-level signals from distributed, labile networks is an intrinsic feature of dlPFC microcircuitry. [Display omitted] • dlPFC intrinsically maintains recurrent activity in the absence of extrinsic inputs • Identical inputs are routed through labile, stochastically assembled ensembles • Stable population-level representations arise from unstable ensembles • Microcircuit excitability covaried with individual cognitive flexibility Using ex vivo Ca2+ imaging in macaque dorsolateral prefrontal cortex, Nolan et al. reveal that recurrent activity propagates through unstable, stochastically assembled ensembles to produce stable population-level events. Network excitability covaried with set-shifting performance, suggesting that these features may underlie dlPFC's role in cognition. [ABSTRACT FROM AUTHOR]

Neurodevelopmental disorders (NDDs) affect 4.7% of the global population and are associated with delays in brain development and a spectrum of impairments that can lead to lifelong disability and even mortality. Identification of biomarkers for accurate diagnosis and medications for effective treatment are lacking, in part due to the historical use of preclinical model systems that do not translate well to the clinic for neurological disorders, such as rodents and heterologous cell lines. Human-induced pluripotent stem cells (hiPSCs) are a promising in vitro system for modeling NDDs, providing opportunities to understand mechanisms driving NDDs in human neurons. Functional assays, including patch clamping, multielectrode array, and imaging-based assays, are popular tools employed with hiPSC disease models for disease investigation. Recent progress in machine learning (ML) algorithms also presents unprecedented opportunities to advance the NDD research process. In this review, we compare two-dimensional and three-dimensional hiPSC formats for disease modeling, discuss the applications of functional assays, and offer insights on incorporating ML into hiPSC-based NDD research and drug screening. [ABSTRACT FROM AUTHOR]

The primary method of treatment for patients suffering from drug-resistant focal-onset epilepsy is resective surgery, which adversely impacts neurocognitive function. Radio frequency (RF) ablation and laser ablation are the methods with the most promise, achieving seizure-free rates similar to resection but with less negative impact on neurocognitive function. However, there remains a number of concerns and open technical questions about these two methods of thermal ablation, with the primary ones: (1) heating; (2) hemorrhage and bleeding; and (3) poor directionality. Irreversible electroporation (IRE) is a proven method of focal ablation, which circumvents all three of the primary concerns regarding focal RF and laser ablation. Here, we demonstrate the in vivo application of a flexible implant with organic electrodes for focal ablation of epilepsy foci using high-frequency IRE (H-FIRE) in mice. Our results show that local, targeted ablation is possible in the close neighborhood of the electrode, paving the way for the clinical application in the treatment of focal epilepsy. [ABSTRACT FROM AUTHOR]

Simple Summary: Drosophila suzukii is a major pest that damages berries and stone fruits by laying its eggs in ripening fruit still on the plant, in contrast to its sister species Drosophila melanogaster, which lays its eggs on overripe, fermenting fruit on the ground. Both species rely on their sense of smell to find such fruit, but how their brains process these odours and subsequently decide on different hosts is not yet fully understood. In this study, we hypothesised that the differences in behaviour may begin in the antennal lobes, the first brain regions of the olfactory system. Using advanced brain imaging techniques, we investigated how the two species respond to odours of ripe fruit, fermented fruit, leaves, and bacteria. We found structural differences in the antennal lobes and differences in the way the odours are represented in these areas of the brain, while behavioural experiments looking for direct differences in the attractiveness of the tested odours revealed no significant variation between the species. The differences in odour processing could form the basis for alternative species-specific pest control strategies that could reduce dependence on insecticides. Drosophila suzukii severely damages the production of berry and stone fruits in large parts of the world. Unlike D. melanogaster, which reproduces on overripe and fermenting fruits on the ground, D. suzukii prefers to lay its eggs in ripening fruits still on the plants. Flies locate fruit hosts by their odorant volatiles, which are detected and encoded by a highly specialised olfactory system before being translated into behaviour. The exact information-processing pathway is not yet fully understood, especially the evaluation of odour attractiveness. It is also unclear what differentiates the brains of D. suzukii and D. melanogaster to cause the crucial difference in host selection. We hypothesised that the basis for different behaviours is already formed at the level of the antennal lobe of D. suzukii and D. melanogaster by different neuronal responses to volatiles associated with ripe and fermenting fruit. We thus investigated by 3D in vivo two-photon calcium imaging how both species encoded odours from ripe fruits, leaves, fermented fruits, bacteria, and their mixtures in the antennal lobe. We then assessed their behavioural responses to mixtures of ripe and fermenting odours. The neural responses reflect species-dependent shifts in the odour code. In addition to this, morphological differences were also observed. However, this was not directly reflected in different behavioural responses to the odours tested. [ABSTRACT FROM AUTHOR]

Neural information transmission between deep brain nuclei and the cortex is essential for brain function. Currently, high-resolution simultaneous detection of neural information between the deep brain nuclei and the large-scale cortex still poses challenges. We have developed the microelectrode arrays based on the Micro-Electro Mechanical System technology, and modified the electrode surface with nanomaterials to improve the electrode performance. This study combined microelectrode arrays and extended-field-of-view microscopy to achieve simultaneous recording of claustrum (CLA) electrophysiology and wide-field cortical calcium imaging at single-cell resolution. This work investigated the synchronous changes of neural information in CLA and cortex of mice during the whole process from wakefulness to anesthesia and then to wakefulness, and summarized the characteristics of the CLA electrophysiology and cortical calcium signaling under different inhalation anesthesia concentrations. We found the synergy between microscopic spike and local field potential of CLA neurons under deep anesthesia, and the law that high inhalation anesthesia concentration enhanced the synchronization between neurons in CLA and cortex. The combination of microelectrode arrays and extended-field-of-view microscopy also gives a new method for synchronous detection of multimodal and multi-brain region neural information. [ABSTRACT FROM AUTHOR]

False clownfish (Amphiprion ocellaris) employ a hatching strategy regulated by environmental cues, wherein parents provide water flow to encourage embryos to hatch after sunset on the hatching day. Despite previous studies demonstrating the necessity of complete darkness and water agitation for hatching, the regulatory mechanisms underlying these environmental cues remain elusive. This study aimed to investigate how darkness and water agitation affect the secretion of hatching enzymes and the hatching movements of embryos in false clownfish. Assessment of chorion digestion and live imaging of Ca2+ in hatching glands using GCaMP6s, a Ca2+ indicator, revealed that darkness stimulation triggers the secretion of hatching enzymes by increasing Ca2+ levels in hatching gland cells. On the other hand, water agitation primarily stimulated hatching movements in embryos, which led to the rupture of their egg envelopes. These results suggest that changes in light environments following sunset induce embryos to secrete hatching enzymes and that water agitation provided by parents stimulates hatching movements. These responses to environmental cues, light and water agitation, contribute to the rapid and synchronous hatching in false clownfish. [ABSTRACT FROM AUTHOR]

Abstract Calcium plays an important role in regulating various neuronal activities in human brains. Investigating the dynamics of the calcium level in neurons is essential not just for understanding the pathophysiology of neuropsychiatric disorders but also as a quantitative gauge to evaluate the influence of drugs on neuron activities. Accessing human brain tissue to study neuron activities has historically been challenging due to ethical concerns. However, a significant breakthrough in the field has emerged with the advent of utilizing patient-derived human induced pluripotent stem cells (iPSCs) to culture neurons and develop brain organoids. This innovative approach provides a promising modeling system to overcome these critical obstacles. Many robust calcium imaging analysis tools have been developed for calcium activity analysis. However, most of the tools are designed for calcium signal detection only. There are limited choices for in-depth downstream applications, particularly in discerning differences between patient and normal calcium dynamics and their responses to drug treatment obtained from human iPSC-based models. Moreover, end-user researchers usually face a considerable challenge in mastering the entire analysis procedure and obtaining critical outputs due to the steep learning curve associated with these available tools. Therefore, we developed CalciumZero, a user-friendly toolbox to satisfy the unmet needs in calcium activity studies in human iPSC-based 3D-organoid/neurosphere models. CalciumZero includes a graphical user interface (GUI), which provides end-user iconic visualization and smooth adjustments on parameter tuning. It streamlines the entire analysis process, offering full automation with just one click after parameter optimization. In addition, it includes supplementary features to statistically evaluate the impact on disease etiology and the detection of drug candidate effects on calcium activities. These evaluations will enhance the analysis of imaging data obtained from patient iPSC-derived brain organoid/neurosphere models, providing a more comprehensive understanding of the results.

Abstract Calcium imaging allows recording from hundreds of neurons in vivo with the ability to resolve single cell activity. Evaluating and analyzing neuronal responses, while also considering all dimensions of the data set to make specific conclusions, is extremely difficult. Often, descriptive statistics are used to analyze these forms of data. These analyses, however, remove variance by averaging the responses of single neurons across recording sessions, or across combinations of neurons, to create single quantitative metrics, losing the temporal dynamics of neuronal activity, and their responses relative to each other. Dimensionally Reduction (DR) methods serve as a good foundation for these analyses because they reduce the dimensions of the data into components, while still maintaining the variance. Nonnegative Matrix Factorization (NMF) is an especially promising DR analysis method for analyzing activity recorded in calcium imaging because of its mathematical constraints, which include positivity and linearity. We adapt NMF for our analyses and compare its performance to alternative dimensionality reduction methods on both artificial and in vivo data. We find that NMF is well-suited for analyzing calcium imaging recordings, accurately capturing the underlying dynamics of the data, and outperforming alternative methods in common use.

Abstract In recent years, research has progressively increased the importance of considering sex differences in stress and fear memory studies. Many studies have traditionally focused on male subjects, potentially overlooking critical differences with females. Emerging evidence suggests that males and females can exhibit distinct behavioral and neurophysiological responses to stress and fear conditioning. These differences may be attributable to variations in hormone levels, brain structure, and neural circuitry, particularly in regions such as the prefrontal cortex (PFC). In the present study, we explored sex differences in prelimbic cortex (PL) calcium activity in animals submitted to immobilization stress (IMO), fear conditioning (FC), and fear extinction (FE). While no significant sex differences were found in behavioral responses, we did observe differences in several PL calcium activity parameters. To determine whether these results were related to behaviors beyond stress and fear memory, we conducted correlation studies between the movement of the animals and PL activity during IMO and freezing behavior during FC and FE. Our findings revealed a clear correlation between PL calcium activity with movement during stress exposure and freezing behavior, with no sex differences observed in these correlations. These results suggest a significant role for the PL in movement and locomotion, in addition to its involvement in fear-related processes. The inclusion of both female and male subjects is crucial for studies like this to fully understand the role of the PFC and other brain areas in stress and fear responses. Recognizing sex differences enhances our comprehension of brain function and can lead to more personalized and effective approaches in the study and treatment of stress and fear-related conditions.

G protein-coupled receptor 139 (GPR139), a highly conserved orphan receptor, is predominantly expressed in the habenula of vertebrate species. Habenula is an ancient epithalamic structure, which is critical to comprehending adaptive behaviors in vertebrates. We have previously demonstrated the role of GPR139 agonists in fear-associated decision-making processes in zebrafish. However, how GPR139 signaling in the habenula modulates such adaptive behavioral responses remains unsolved. Fish centrally administered with a synthetic antagonist for human GPR139 (NCRW0005-F05) exhibited significant suppression of odorant cue (alarm substance, AS)-induced fear learning in the conditioned place avoidance paradigm. On the other hand, co-treatment with a GPR139 antagonist and a synthetic agonist for human GPR139 (JNJ-63533054) interrupted the fear conditioning process by significantly reducing locomotion during post-conditioning. Calcium imaging of acute brain slices showed a significant increase in peak amplitude of calcium transients in the habenula upon bath application of either a GPR139 antagonist or agonist. Furthermore, KCl-evoked calcium transients were reduced by the GPR139 antagonist and co-treatment of the GPR139 antagonist–agonist. These results suggest that the GPR139 antagonist did not block the inhibitory action of the GPR139 agonist in the decision-making process during the fear-retrieval phase; however, solitarily, it functions in governing the fear consolidation process via activation of the ventral habenula neurons in zebrafish. [ABSTRACT FROM AUTHOR]

Miniature fluorescence microscopes (miniscopes) are one of the most powerful and versatile tools for recording large scale neural activity in freely moving rodents with single cell resolution. Recent advances in the design of genetically encoded calcium indicators (GECIs) allow to target distinct neuronal populations with non-overlapping emission spectral profiles. However, conventional miniscopes are limited to a single excitation, single focal plane imaging, which does not allow to compensate for chromatic aberration and image from two spectrally distinct calcium indicators. In this paper we present an open-source dual channel miniscope capable of simultaneous imaging of genetically or functionally distinct neuronal populations. Chromatic aberrations are corrected using an electrowetting lens (EWL), which allows fast focal plane change between frames. To demonstrate the capabilities of the dual channel miniscope, we labeled layer specific excitatory neurons or inhibitory interneurons in the medial prefrontal cortex (mPFC) with a red fluorescence protein, and simultaneously imaged neural activity of distinct neuronal populations of freely moving mice via a green GECI. [ABSTRACT FROM AUTHOR]

The hippocampus forms unique neural representations for distinct experiences, supporting the formation of different memories. 1,2,3,4,5,6 Hippocampal representations gradually change over time as animals repeatedly visit the same familiar environment ("representational drift"). 7,8,9,10,11,12 Such drift has also been observed in other brain areas, such as the parietal, 13,14 visual, 15,16,17 auditory, 18,19 and olfactory 20 cortices. While the underlying mechanisms of representational drift remain unclear, a leading hypothesis suggests that it results from ongoing learning processes. 20,21,22 According to this hypothesis, because the brain uses the same neural substrates to support multiple distinct representations, learning of novel stimuli or environments leads to changes in the neuronal representation of a familiar one. If this is true, we would expect drift in a given environment to increase following new experiences in other, unrelated environments (i.e., off-context experiences). To test this hypothesis, we longitudinally recorded large populations of hippocampal neurons in mice while they repeatedly visited a familiar linear track over weeks. We introduced off-context experiences by placing mice in a novel environment for 1 h after each visit to the familiar track. Contrary to our expectations, these novel episodes decreased place cells' representational drift. Our findings are consistent with a model in which representations of distinct memories occupy different areas within the neuronal activity space, and the drift of each of them within that space is constrained by the area occupied by the others. [Display omitted] • Ca²⁺ imaging was used to quantify representational drift in a familiar environment • Mice were exposed to novel enriched environments to boost hippocampal learning • Off-context novel experiences constrain hippocampal drift rather than accelerate it • Off-context novel experiences specifically affect drift in spatial tuning A leading hypothesis suggests that representational drift in a given context stems from ongoing learning processes, such as those induced by novelty in other contexts. However, Elyasaf et al. find evidence to the contrary: novel experiences constrain hippocampal representational drift rather than accelerating it. [ABSTRACT FROM AUTHOR]

Coordinated activity of neuronal ensembles is a basis for information processing in the brain. Recent development of miniscope imaging technology enabled recordings of neuronal circuits activity in vivo in freely behaving animals. Acute stress is believed to affect various hippocampal functions, especially memory. In the current study, we utilized miniscope imaging to investigate the hippocampal neuronal circuits properties in a mouse as function of time and immediately in response to an acute stress, induced by passive restraint, 3 h and 10 days after. Comprehensive quantitative analysis of network activity changes at the neuronal ensembles level revealed highly stable neuronal activity parameters, which exhibited a rapid and robust shift in response to acute stress stimulation. This shift was accompanied by the restructuring of the pairwise-correlated neuronal pairs. Remarkably, we discovered that ensembles activity characteristics returned to the initial state following recovery period, demonstrating hippocampal homeostatic stability at the neuronal circuits level. Obtained results provide an evidence about hippocampal neuronal ensembles activity in response to acute stress over time. [ABSTRACT FROM AUTHOR]

There is mounting evidence that the cerebellum impacts hippocampal functioning, but the impact of the cerebellum on hippocampal interneurons remains obscure. Using miniscopes in freely behaving male and female mice, we found optogenetic stimulation of Purkinje cells alters the calcium activity of a large percentage of CA1 interneurons. This includes both increases and decreases in activity. Remarkably, this bidirectional impact occurs in a coordinated fashion, in line with interneurons' functional properties. Specifically, CA1 interneurons activated by cerebellar stimulation are commonly locomotion-active, while those inhibited by cerebellar stimulation are commonly rest-active interneurons. We additionally found that subsets of CA1 interneurons show altered activity during object investigations. Importantly, these interneurons also show coordinated modulation by cerebellar stimulation: CA1 interneurons that are activated by cerebellar stimulation are more likely to be activated, rather than inhibited, during object investigations, while interneurons that show decreased activity during cerebellar stimulation show the opposite profile. We examined two different stimulation locations (IV/V vermis or simplex) and two different stimulation approaches (7 Hz or a single 1 s light pulse)--in all cases, the cerebellum induces similar coordinated CA1 interneuron changes congruent with an explorative state. Overall, our data show that CA1 interneurons are impacted by cerebellar manipulation in a bidirectional and coordinated fashion and are therefore likely to play an important role in cerebello-hippocampal communication. [ABSTRACT FROM AUTHOR]

3,4‐methylenedioxymethamphetamine (MDMA), a commonly abused recreational drug, induces prosocial effects such as increased sociability and empathy. The nucleus accumbens (NAc) has been suggested to play a crucial role in these MDMA‐mediated prosocial effects. However, the relationship between social behavior and NAc neural activity, and the effects of MDMA on this relationship, remain unknown. In this study, we measured NAc neural activity using fiber photometry and classified the behaviors of mice at times of transient increases in NAc neural activity during the social approach test (SAT). We found that NAc neural activity transiently increased at the onset of turning toward and sniffing novel mice during the SAT, although the frequency of turning was relatively low. We then examined the effects of MDMA on behavior and NAc neural activity and found that MDMA decreased the duration of sniffing per bout but did not alter NAc neural activity at the onset of turning toward or sniffing novel mice. These results suggest that MDMA does not affect the transient increase in NAc neural activity at the onset of social behaviors. [ABSTRACT FROM AUTHOR]

The brain's ability to integrate sensory and motor information allows us to maintain a sense of orientation in space, a process in which head‐direction cells play a key role. While these neurons have been studied extensively in mammals, their presence and function in non‐mammalian species remain less understood. Here, I summarize the research work for my PhD thesis, where we explore the interpeduncular nucleus (IPN) in zebrafish, a lesser known brain region, using whole‐brain electron microscopy and calcium imaging techniques. We identified a novel population of unipolar neurons, with their activity exhibiting a dynamic, rotational pattern during head movements, even in the absence of sensory cues. This population mirrors the functionality of head‐direction cells observed in mammals, suggesting a conserved mechanism for spatial orientation across vertebrates. Our findings reveal the potential of the zebrafish IPN as a vertebrate model for studying ring attractor networks, a theoretical framework previously used to explain head‐direction cell activity. These results pave the way for future research on how motor and sensory signals converge in the vertebrate brain to maintain spatial orientation. [ABSTRACT FROM AUTHOR]

Long-term depression (LTD) of synaptic transmission at parallel fiber (PF)-Purkinje cell (PC) synapses plays an important role in cerebellum-related motor coordination and learning. LTD is induced by the conjunction of PF stimulation and climbing fiber (CF) stimulation or somatic PC depolarization, while long-term potentiation (LTP) is induced by PF stimulation alone. Therefore, it is considered that different types of stimulation induce different types of synaptic plasticity. However, we found that a small number of conjunctive stimulations (PF + somatic depolarization of PC) induced LTP, but did not induce LTD of a small size. This LTP was not associated with changes in paired-pulse ratio, suggesting postsynaptic origin. Additionally this LTP was dependent on nitric oxide. This LTP was also induced by a smaller number of physiological conjunctive PF and CF stimuli. These results suggested that a larger number or longer period of conjunctive stimulation is required to induce LTD by overcoming LTP. Ca2+ transients at the PC dendritic region was measured by calcium imaging during LTD-inducing conjunctive stimulation. Peak amplitude of Ca2+ transients increased gradually during repetitive conjunctive stimulation. Instantaneous peak amplitude was not different between the early phase and late phase, but the average amplitude was larger in the later phase than in the early phase. These results show that LTD overcomes LTP, and increased Ca2+ integration or a number of stimulations is required for LTD induction. [ABSTRACT FROM AUTHOR]

In vitro neuronal culture is an important research platform in cellular and network neuroscience. However, neurons cultured on a homogeneous scaffold form dense, randomly connected networks and display excessively synchronized activity; this phenomenon has limited their applications in network‐level studies, such as studies of neuronal ensembles, or coordinated activity by a group of neurons. Herein, polydimethylsiloxane‐based microfluidic devices are developed to create small neuronal networks exhibiting a hierarchically modular structure resembling the connectivity observed in the mammalian cortex. The strength of intermodular coupling is manipulated by varying the width and height of the microchannels that connect the modules. Neuronal activity recording via calcium imaging shows that the spontaneous activity in networks with smaller microchannels (2.2–5.5 µm2) has lower synchrony and exhibits a threefold variety of neuronal ensembles. Optogenetic stimulation demonstrates that a reduction in intermodular coupling enriches evoked neuronal activity patterns and that repeated stimulation induces plasticity in neuronal ensembles in these networks. These findings suggest that cell engineering technologies based on microfluidic devices enable in vitro reconstruction of the intricate dynamics of neuronal ensembles, thus providing a robust platform for studying neuronal ensembles in a well‐defined physicochemical environment. [ABSTRACT FROM AUTHOR]

Traumatic brain injury (TBI) affects neural function at the local injury site and also at distant, connected brain areas. However, the real‐time neural dynamics in response to injury and subsequent effects on sensory processing and behaviour are not fully resolved, especially across a range of spatial scales. We used in vivo calcium imaging in awake, head‐restrained male and female mice to measure large‐scale and cellular resolution neuronal activation, respectively, in response to a mild/moderate TBI induced by focal controlled cortical impact (CCI) injury of the motor cortex (M1). Widefield imaging revealed an immediate CCI‐induced activation at the injury site, followed by a massive slow wave of calcium signal activation that travelled across the majority of the dorsal cortex within approximately 30 s. Correspondingly, two‐photon calcium imaging in the primary somatosensory cortex (S1) found strong activation of neuropil and neuronal populations during the CCI‐induced travelling wave. A depression of calcium signals followed the wave, during which we observed the atypical activity of a sparse population of S1 neurons. Longitudinal imaging in the hours and days after CCI revealed increases in the area of whisker‐evoked sensory maps at early time points, in parallel to decreases in cortical functional connectivity and behavioural measures. Neural and behavioural changes mostly recovered over hours to days in our M1‐TBI model, with a more lasting decrease in the number of active S1 neurons. Our results in unanaesthetized mice describe novel spatial and temporal neural adaptations that occur at cortical sites remote to a focal brain injury. [ABSTRACT FROM AUTHOR]

Feedback at the photoreceptor synapse is the first neuronal circuit computation in vision, which influences downstream activity patterns within the visual system. Yet, the identity of the feedback signal and the mechanism of synaptic transmission are still not well understood. Here, we combined perturbations of cell-type-specific genes of mouse horizontal cells with two-photon imaging of the result of light-induced feedback in cones and showed that the electrogenic bicarbonate transporter Slc4a5, but not the electroneutral bicarbonate transporter Slc4a3, both expressed specifically in horizontal cells, is necessary for horizontal cell-to-cone feedback. Pharmacological blockage of bicarbonate transporters and buffering pH also abolished the feedback but blocking sodium-proton exchangers and GABA receptors did not. Our work suggests an unconventional mechanism of feedback at the first visual synapse: changes in horizontal cell voltage modulate bicarbonate transport to the cell, via Slc4a5, which leads to the modulation of feedback to cones. [Display omitted] • Feedback from horizontal cells to cones was measured by imaging calcium in cones • Slc4a5 is an electrogenic Na+-HCO 3 − cotransporter specific to horizontal cells • Knocking out or down Slc4a5 abolished horizontal cell-to-cone feedback • Blocking HCO 3 − transporters and buffering pH also abolished feedback Molecular mechanisms of horizontal cell-to-cone feedback in the retina have been under debate. Morikawa et al. show that Slc4a5, an electrogenic sodium-bicarbonate cotransporter specific to horizontal cells, is crucial for feedback. They suggest that Slc4a5, through its voltage-dependent transport of bicarbonate, regulates pH in the synaptic cleft that mediates feedback. [ABSTRACT FROM AUTHOR]

Calcium imaging allows recording from hundreds of neurons in vivo with the ability to resolve single cell activity. Evaluating and analyzing neuronal responses, while also considering all dimensions of the data set to make specific conclusions, is extremely difficult. Often, descriptive statistics are used to analyze these forms of data. These analyses, however, remove variance by averaging the responses of single neurons across recording sessions, or across combinations of neurons, to create single quantitative metrics, losing the temporal dynamics of neuronal activity, and their responses relative to each other. Dimensionally Reduction (DR) methods serve as a good foundation for these analyses because they reduce the dimensions of the data into components, while still maintaining the variance. Nonnegative Matrix Factorization (NMF) is an especially promising DR analysis method for analyzing activity recorded in calcium imaging because of its mathematical constraints, which include positivity and linearity. We adapt NMF for our analyses and compare its performance to alternative dimensionality reduction methods on both artificial and in vivo data. We find that NMF is well-suited for analyzing calcium imaging recordings, accurately capturing the underlying dynamics of the data, and outperforming alternative methods in common use. [ABSTRACT FROM AUTHOR]

Background: Hypofunction of the glutamate system in the brain is one of the pathophysiological hypotheses for schizophrenia. Accumulating animal and clinical studies show that sarcosine (N-methylglycine), a glycine transporter-1 inhibitor, is effective in ameliorating the negative and cognitive symptoms of schizophrenia. The aims of the present study were to observe the effects of sarcosine on neuronal activity in the dorsal CA1 (dCA1) hippocampal neurons within an NMDA receptor hypofunction model induced by MK801. Methods: We applied in vivo calcium imaging to observe the dynamics of fluorescence from the dCA1 hippocampal neurons when the mice were exploring in an open field. Using this tool, we directly measured and compared neuronal properties between sarcosine-treated and untreated mice. At the same time, the physiological function of the neurons was also quantified by measuring their place fields. Results: Our data demonstrated that MK-801 (0.2 mg/kg) diminished the fluorescence intensity of dCA1 neurons that had been genetically modified with a calcium indicator. MK-801 also significantly increased the correlation coefficient between the fluorescence dynamics of pairs of cells, a feature that may be linked to the symptom of disorganization in human patients with schizophrenia. The spatial correlations of place fields in the mice were impaired by MK-801 as well. Injected sarcosine (500 mg or 1000 mg/kg) significantly alleviated the abovementioned abnormalities. Conclusions: Our data provide evidence to support the use of sarcosine to alleviate symptoms of schizophrenia, especially hippocampus-related functions. [ABSTRACT FROM AUTHOR]

INTRODUCTION: It is unclear how early neuronal deficits occur in tauopathies, if these are associated with changes in neuronal network activity, and if they can be alleviated with therapies. METHODS: To address this, we performed in vivo two‐photon Ca2+ imaging in tauopathy mice at 6 versus 12 months, compared to controls, and treated the younger animals with a tau antibody. RESULTS: Neuronal function was impaired at 6 months but did not deteriorate further at 12 months, presumably because cortical tau burden was comparable at these ages. At 6 months, neurons were mostly hypoactive, with enhanced neuronal synchrony, and had dysregulated responses to stimulus. Ex vivo, electrophysiology revealed altered synaptic transmission and enhanced excitability of motor cortical neurons, which likely explains the altered network activity. Acute tau antibody treatment reduced pathological tau and gliosis and partially restored neuronal function. DISCUSSION: Tauopathies are associated with early neuronal deficits that can be attenuated with tau antibody therapy. Highlights: Neuronal hypofunction in awake and behaving mice in early stages of tauopathy.Altered network activity disrupted local circuitry engagement in tauopathy mice.Enhanced neuronal excitability and altered synaptic transmission in tauopathy mice.Tau antibody acutely reduced soluble phospho‐tau and improved neuronal function. [ABSTRACT FROM AUTHOR]

Acidic stress is a formidable environmental factor that exerts adverse effects on plant growth and development, ultimately leading to a potential reduction in agricultural productivity. A low pH triggers Ca2+ influx across the plasma membrane (PM), eliciting distinct responses under various acidic pH levels. However, the underlying mechanisms by which Arabidopsis plant cells generate stimulus-specific Ca2+ signals in response to acidic stress remain largely unexplored. The experimentally induced stimulus may elicit spikes in cytosolic free Ca2+ concentration ([Ca2+]i) spikes or complex [Ca2+]i oscillations that persist for 20 min over a long-term of 24 h or even several days within the plant cytosol and chloroplast. This study investigated the increase in [Ca2+]i under a gradient of low pH stress ranging from pH 3.0 to 6.0. Notably, the peak of [Ca2+]i elevation was lower at pH 4.0 than at pH 3.0 during the initial 8 h, while other pH levels did not significantly increase [Ca2+]i compared to low acidic stress conditions. Lanthanum chloride (LaCl3) can effectively suppress the influx of [Ca2+]i from the apoplastic to the cytoplasm in plants under acid stress, with no discernible difference in intracellular calcium levels observed in Arabidopsis. Following 8 h of acid treatment in the darkness, the intracellular baseline Ca2+ levels in Arabidopsis were significantly elevated when exposed to low pH stress. A moderately low pH, specifically 4.0, may function as a spatial-temporal input into the circadian clock system. These findings suggest that acid stimulation can exert a continuous influence on intracellular calcium levels, as well as plant growth and development. [ABSTRACT FROM AUTHOR]

Background/Objectives: Adiponectin, the most abundant peptide hormone secreted by adipocytes, is a well-known homeostatic factor regulating lipid metabolism and insulin sensitivity. It has been shown that the adiponectin receptor agonist AdipoRon selectively enhances cellular responses to fatty acids in human taste cells, and adiponectin selectively increases taste behavioral responses to intralipid in mice. However, the molecular mechanism underlying the physiological effects of adiponectin on fat taste in mice remains unclear. Conclusions: Here we define AdipoR1 as the mediator responsible for the enhancement role of adiponectin/AdipoRon on fatty acid-induced responses in mouse taste bud cells. Methods and Results: Calcium imaging data demonstrate that AdipoRon enhances linoleic acid-induced calcium responses in a dose-dependent fashion in mouse taste cells isolated from circumvallate and fungiform papillae. Similar to human taste cells, the enhancement role of AdipoRon on fatty acid-induced responses was impaired by co-administration of an AMPK inhibitor (Compound C) or a CD36 inhibitor (SSO). Utilizing Adipor1-deficient animals, we determined that the enhancement role of AdipoRon/adiponectin is dependent on AdipoR1, since AdipoRon/adiponectin failed to increase fatty acid-induced calcium responses in taste bud cells isolated from these mice. Brief-access taste tests were performed to determine whether AdipoRon's enhancement role was correlated with any differences in taste behavioral responses to fat. Although AdipoRon enhances the cellular responses of taste bud cells to fatty acids, it does not appear to alter fat taste behavior in mice. However, fat-naïve Adipor1−/− animals were indifferent to increasing concentrations of intralipid, suggesting that adiponectin signaling may have profound effects on the ability of mice to detect fatty acids in the absence of previous exposure to fatty acids and fat-containing diets. [ABSTRACT FROM AUTHOR]

Thermogenetics is a promising neuromodulation technique based on the use of heat-sensitive ion channels. However, on the way to its clinical application, a number of questions have to be addressed. First, to avoid immune response in future human applications, human ion channels should be studied as thermogenetic actuators. Second, heating levels necessary to activate these channels in vivo in brain tissue should be studied and cytotoxicity of these temperatures addressed. Third, the possibility and safety of chronic neuromodulation has to be demonstrated. In this study, we present a comprehensive framework for thermogenetic neuromodulation in vivo using the thermosensitive human ion channel hTRPV1. By targeting hTRPV1 expression to excitatory neurons of the mouse brain and activating them within a non-harmful temperature range with a fiber-coupled infrared laser, we not only induced neuronal firing and stimulated locomotion in mice, but also demonstrated that thermogenetics can be employed for repeated neuromodulation without causing evident brain tissue injury. Our results lay the foundation for the use of thermogenetic neuromodulation in brain research and therapy of neuropathologies. [ABSTRACT FROM AUTHOR]

A comprehensive understanding of physio-pathological processes necessitates non-invasive intravital three-dimensional (3D) imaging over varying spatial and temporal scales. However, huge data throughput, optical heterogeneity, surface irregularity, and phototoxicity pose great challenges, leading to an inevitable trade-off between volume size, resolution, speed, sample health, and system complexity. Here, we introduce a compact real-time, ultra-large-scale, high-resolution 3D mesoscope (RUSH3D), achieving uniform resolutions of 2.6 × 2.6 × 6 μm3 across a volume of 8,000 × 6,000 × 400 μm3 at 20 Hz with low phototoxicity. Through the integration of multiple computational imaging techniques, RUSH3D facilitates a 13-fold improvement in data throughput and an orders-of-magnitude reduction in system size and cost. With these advantages, we observed premovement neural activity and cross-day visual representational drift across the mouse cortex, the formation and progression of multiple germinal centers in mouse inguinal lymph nodes, and heterogeneous immune responses following traumatic brain injury—all at single-cell resolution, opening up a horizon for intravital mesoscale study of large-scale intercellular interactions at the organ level. [Display omitted] • RUSH3D allows long-term, fast 3D imaging at centimeter-scale and single-cell resolution • Compact, high-fidelity mesoscale imaging is achieved in native intravital environments • Intercellular interactions across multiple mouse germinal centers are observed in vivo • Cortex-wide multisensory, locomotion neural activities and immune responses are explored in mice RUSH3D, with the integration of multiple computational imaging methods based on a scanning light-field framework, facilitates long-term, high-speed, centimeter-wide mesoscale 3D imaging at single-cell resolution in a compact system for broad practical applications in understanding large-scale intercellular dynamics at the mammalian organ level. We have demonstrated cortex-wide large-population neural recording with cross-day registration during multisensory input, the formation and progression of multiple germinal centers in mouse inguinal lymph nodes, and cortex-wide heterogeneous immune responses after traumatic brain injury. [ABSTRACT FROM AUTHOR]