bioRxiv Subject Collection: Neuroscience's Journal

Maintenance of neuronal TDP-43 expression requires axonal lysosome transport
TDP-43 mislocalization and pathology occurs across a range of neurodegenerative diseases, but the pathways that modulate TDP-43 in neurons are not well understood. We generated a Halo-TDP-43 knock-in iPSC line and performed a genome-wide CRISPR interference FACS-based screen to identify modifiers of TDP-43 levels in neurons. A meta-analysis of our screen and publicly available screens identified both specific hits and pathways present across multiple screens, the latter likely responsible for generic protein level maintenance. We identified BORC, a complex required for anterograde lysosome transport, as a specific modifier of TDP-43 protein, but not mRNA, levels in neurons. BORC loss led to longer half-life of TDP-43 and other proteins, suggesting lysosome location is required for proper protein turnover. As such, lysosome location and function are crucial for maintaining TDP-43 protein levels in neurons.

Neuronal activity regulating the dauer entry decision in Caenorhabditis elegans
The model nematode Caenorhabditis elegans can choose between two alternative developmental trajectories. Larvae can either become reproductive adults or, under conditions of crowding or low food availability, enter a long-term, stress-resistant diapause known as the dauer stage. Previous studies showed that chemical signals from a secreted larval pheromone promote the dauer trajectory, and that their influence can be antagonised by increased availability of microbial food. The decision is known to be under neuronal control, involving both sensory and interneurons. To make an accurate decision, larvae must collect and compare complex patterns of environmental input over around 25 hours of development. The full composition of this circuit and the algorithm for decision-making are unknown. Here, we used cell-specific chemical silencing to systematically perturb several sensory and interneurons to further elucidate circuit composition. Our results suggest a role for gas-sensing neurons in regulating dauer entry. In addition, we quantitatively characterized the neuronal responses to food and pheromone inputs by measuring calcium traces from ASI and AIA neurons. We found that calcium in ASI increases linearly in response to food, and similarly decreases in response to pheromone, revealing a cellular site of antagonism between these key chemical inputs. Notably, the ASI response persists well beyond removal of the food stimulus, thus encoding a memory of recent food exposure. In contrast, AIA reports instantaneous food availability, and is unaffected by pheromone. We discuss how these findings may inform our understanding of this long-term decision-making process.

Emergent glutamate & dopamine dysfunction in VPS35(D620N) knock-in mice and rapid reversal by LRRK2 inhibition
The D620N variant in Vacuolar Protein Sorting 35 (VPS35) causes autosomal-dominant, late-onset Parkinson's disease. VPS35 is a core subunit of the retromer complex that canonically recycles transmembrane cargo from sorting endosomes. Although retromer cargoes include many synaptic proteins, VPS35's neuronal functions are poorly understood. To investigate the consequences of the Parkinson's mutation, striatal neurotransmission was assessed in 1-, 3- & 6-month-old VPS35 D620N knock-in (VKI) mice. Spontaneous and optogenetically-evoked corticostriatal glutamate transmission was increased in VKI striatal spiny projection neurons by 6 months, when total striatal glutamate release, quantified by iGluSnFR imaging, showed similarities to wild-type. dLight imaging revealed robust increases in VKI striatal dopamine release by 6 months, which were reversed with acute ex vivo leucine-rich repeat kinase 2 (LRRK2) inhibition. We conclude that increased glutamate and dopamine transmission in VKI mice progressively emerges in young-adulthood, and that dopamine dysfunction is likely the result of sustained, rapidly-reversible, LRRK2 hyperactivity.

Mental Effort and Counterfactuals Modulate Language Understanding: ERP Evidence in Older Adults
The relationship between language and physical effort in older adults is a field that is scarcely explored in the literature associated with embodiment. An electrophysiological experiment was conducted to explore the modulation of two linguistic contexts: factual and counterfactual, in relation to physical and mental effort using electrophysiological components. 27 older adults (M = 70.34 years, SD = 4.82, 15 women and 12 men) read sentences on a computer screen and responded to an activation test. The results indicate that the linguistic, factual, and counterfactual contexts, as well as the embodiment parameter of mental effort modulate the understanding of language and participate with variable preponderance in different time windows. Furthermore, counterfactuality seems to facilitate the processing of high mental effort, and both factual and counterfactual language elicit the N400 component. These findings contribute to the growing body of research on embodied cognition by providing novel insights into the nuances of cognitive demands involved in language processing in aging population, paving the way for developing targeted interventions aimed at improving communication and cognitive well-being in older adults.

A novel method for harmonization of PET image spatial resolution without phantoms
The most common approach for estimating the spatial resolution of PET images in multi-center studies typically uses Hoffman phantom data as a surrogate. Specifically, the phantom-based matching resolution approach assumes that scanned phantom PET images are well approximated by a ground truth, noise-free digital phantom convolved with a Gaussian kernel of unknown size. The size of the kernel is then estimated by an exhaustive search on the amount of blurring needed to match the smoothed digital phantom to a particular scanned phantom image. Unfortunately, Hoffman phantom images may not always be readily available, and phantom-based approaches may yield sub-optimal results. We propose a new, computational approach that allows estimation of spatial resolution directly from the PET image itself. We generalized the so-called logarithmic intensity plots method to the 3D case to perform a spatial resolution estimation in both axial and in-plane directions of the PET images. The proposed approach was applied to two different cohorts. The first cohort consisted of [18F]florbetapir amyloid PET images and matching phantoms coming from a Phase II clinical trial and includes different scanner models and/or orientation and grid reconstructions. The second cohort included beta-amyloid, FDG and tau PET images from the Alzheimer Disease Neuroimaging Initiative (ADNI) study. We obtained in-plane and axial resolution estimators that vary between 3.5 mm and 8.5 mm for both PET and matching phantom images. In both cases, we obtained small across-subject variability in groups of images sharing the same PET scanner model and reconstruction parameters. For human PET images, we also obtained a strong cross-tracer and longitudinal consistency in the spatial resolution estimators. Our novel approach does not only eliminate the need for surrogate brain phantom data, but also provides a general framework that can be applied to a wide range of tracers and other image modalities, such as SPECT.

Linking microscopy to diffusion MRI with degenerate biophysical models: an application of the Bayesian EstimatioN of CHange (BENCH) framework
Biophysical modelling of diffusion MRI (dMRI) is used to non-invasively estimate microstructural features of tissue, particularly in the brain. However, meaningful description of tissue requires many unknown parameters, resulting in a model that is often ill-posed. The Bayesian EstimatioN of CHange (BENCH) framework was specifically designed to circumvent parameter fitting for ill-conditioned models when one is simply interested in interpreting signal changes related to some variable of interest. To understand the biological underpinning of some observed change in MR signal between different conditions, BENCH predicts which model parameter, or combination of parameters, best explains the observed change, without having to invert the model. BENCH has been previously used to identify which biophysical parameters could explain group-wise dMRI signal differences (e.g. patients vs. controls); here, we adapt BENCH to interpret dMRI signal changes related to continuous variables. We investigate how parameters from the dMRI standard model of white matter, with an additional sphere compartment to represent glial cell bodies, relate to tissue microstructure quantified from histology. We validate BENCH using synthetic dMRI data from numerical simulations. We then apply it to ex-vivo macaque brain data with dMRI and microscopy metrics of glial density, axonal density, and axonal dispersion in the same brain. We found that (i) increases in myelin density are primarily associated with an increased intra-axonal volume fraction and (ii) changes in the orientation dispersion derived from myelin microscopy are linked to variations in the orientation dispersion index. Finally, we found that the dMRI signal is sensitive to changes in glial cell soma in the WM, but that no parameter in the extended standard model was able to explain this observed signal change, suggesting model inadequacy.

Functional classification of tauopathy strains reveals the role of protofilament core residues.
Distinct tau amyloid assemblies underlie diverse tauopathies but defy rapid classification. Cell and animal experiments indicate tau functions as a prion, as different strains propagated in cells cause unique, transmissible neuropathology after inoculation. Strain amplification requires compatibility of the monomer and amyloid template. We used cryo-EM to study one cell-based YFP-tagged strain, resolving its amyloid nature. We then used sequential alanine (Ala) substitution (scan) within tau repeat domain (RD) to measure incorporation to pre-existing tau RD-YFP aggregates. This robustly discriminated strains, defining sequences critical for monomer incorporation. We then created 3R/4R or 4R WT RD (aa 246-408) biosensors. Ala scan of recombinant tau seeds with the Alzheimer's Disease fold matched that of AD homogenate. We scanned 22 brain lysates comprising 4 tauopathies. This clustered cases by neuropathological syndrome, revealed the role of amino acids in protofilament folds, and allowed strain discrimination based on amino acid requirements for prion replication.

Oddball evoked deviant responses reflect complex context dependent expectations in mouse V1
Evoked responses in the mouse primary visual cortex can be modulated by the temporal context in which visual inputs are presented. Oddball stimuli embedded in a sequence of regularly repeated visual elements have been shown to drive relatively large deviant responses, a finding that is generally consistent with the theory that cortical circuits implement a form of predictive coding. These results can be confounded by short-term adaptation effects, however, that make interpretation difficult. Here we use various forms of the oddball paradigm to disentangle temporal and ordinal components of the deviant response, showing that it is a complex phenomenon affected by temporal structure, ordinal expectation, and event frequency. Specifically, we use visually evoked potentials to show that deviant responses occur over a large range of time, lead to long-term plasticity in some cases, cannot be explained by a simple adaptation model, scale with predictability, and are modulated by violations of both first and second-order sequential expectations.

Quantitative 2D J-resolved metabolite-cycled semiLASER spectroscopy of metabolites and macromolecules in the human brain at 9.4 T
Purpose: While two-dimensional (2D) in vivo spectroscopy yields rich information and has been successfully used in clinical trials, it requires a localization scheme that minimizes the impact of chemical shift displacement on J-coupling evolution, a robust frequency drift correction and dedicated processing and quantification methods. Considering these needs this study demonstrates a novel data acquisition and an analysis pipeline to quantify 16 metabolites in mmol/kg in the human brain using a 2D J-resolved metabolite-cycled (MC) semiLASER localization sequence at 9.4 T in the human brain. Methods: Metabolite spectra were acquired in vivo using the newly developed J-resolved MC semiLASER localization sequence with maximum echo sampling (MES) at 9.4 T. In order to account for the underlying macromolecular (MM) spectra in the acquired metabolite spectra, J-resolved MM spectra were acquired using a double inversion recovery (DIR) J-resolved MC semiLASER. Spectral fitting was performed with ProFit 2.0 using a simulated basis set from VesPA tailored to 2D J-resolved semiLASER with MES. Finally, metabolite concentrations were calculated using internal water referencing. Results: Tissue concentrations for 16 metabolites in mmol/kg are reported after correcting for number of protons, tissue content, and relaxation effects of both water and metabolites at 9.4T. Quantification results of spectra considering 8 and 2 averages per TE did not show any significant differences. Conclusion: 2D spectra of metabolites acquired at 9.4T and 2D MMs acquired at any field strength are presented for the first time. Basis set simulation and quantification of metabolites for metabolite spectra acquired using maximum-echo-sampled 2D J-resolved semiLASER was performed for the first time. The sensitivity in the detection of J-coupled metabolites such as glutamine, glucose or lactate. At ultra-high field, the acquisition duration of 2D MRS can be also substantially reduced since only a very low number of averages per TE are needed.

Biologically grounded brain-on-chip model identifies selective topographic reorganization within hyperexcitable neural networks
Connectomics has revolutionized our understanding of brain function by emphasizing the importance of neural networks and their topographical organization. Corticostriatal circuits, which play a critical role in cognition and emotion, follow a precise topographic architecture essential for integrating and processing cortical information within the basal ganglia. Disruptions to this connectivity are often implicated in neurodevelopmental and psychiatric disorders such as obsessive compulsive disorders, schizophrenia, epilepsy, and autism spectrum disorders. However, studying network disruptions in vivo presents significant challenges due to their intricate architecture and early developmental onset. To address this, we employed a brain-on-chip microfluidic platform to recreate a biologically relevant model of topographically organized corticostriatal networks. By mimicking the directional control of neuronal projections using Tesla valve-inspired microchannels, we demonstrate that genetic perturbations affecting neuronal excitability during development lead to selective alterations of local versus long-range network topology, resulting in the formation of new convergent nodes. This model offers critical insights into how early perturbations contribute to circuit-specific pathologies, providing a valuable tool for understanding neurodevelopmental disorders and advancing therapeutic strategies.

Dopamine activity encodes the changing valence of the same stimulus in conditioned taste aversion paradigms
Mesolimbic dopamine encoding of non-contingent rewards and reward-predictive cues has been well established. Considerable debate remains over how mesolimbic dopamine responds to aversion and in the context of aversive conditioning. Inconsistencies may arise from the use of aversive stimuli that are transduced along different neural paths relative to reward or the conflation of responses to avoidance and aversion. Here, we made intraoral infusions of sucrose and measured how dopamine and behavioral responses varied to the changing valence of sucrose. Pairing intraoral sucrose with malaise via injection of lithium chloride (LiCl) caused the development of a conditioned taste aversion (CTA), which rendered the typically rewarding taste of sucrose aversive upon subsequent re-exposure. Following CTA formation, intraoral sucrose suppressed the activity of ventral tegmental area dopamine neurons (VTADA) and nucleus accumbens (NAc) dopamine release. This pattern of dopamine signaling after CTA is similar to intraoral infusions of innately aversive quinine and contrasts with that to sucrose when it was novel or not paired with LiCl. Dopamine responses were negatively correlated with behavioral reactivity to intraoral sucrose and predicted home cage sucrose preference. Further, dopamine responses scaled with the strength of the CTA, which was increased by repeated LiCl pairings and weakened through extinction. Thus, the findings demonstrate differential dopamine encoding of the same taste stimulus according to its valence, which is aligned to distinct behavioral responses.

Dissociable after-effects of prosocial acts: Effort is costly for others but valued for self
Prosocial behavior requires effort, yet people are often reluctant to exert effort for others' benefit. However, the manner in which effort exertion affects subsequent reward evaluation during prosocial acts remains elusive. Here, we leveraged the temporal precision of electroencephalography, along with a paradigm that independently manipulated effort and reward for self and another person, to uncover the neural mechanism underlying the reward after-effect of effort expenditure during prosocial acts. We found a dissociable reward after-effect between self-benefitting and other-benefitting effort. When the beneficiary was oneself, the reward positivity (RewP) became more positive as effort increased, indicating an effort-enhancement effect. In contrast, when the beneficiary was others, the RewP became less positive as effort increased, demonstrating an effort-discounting effect. Moreover, this dissociation occurred only when reward system was activated and was independent of performance evaluation. Our finding provides novel insights into how prior effort expenditure shape reward evaluation during prosocial behavior.

Distinct expression profile reveals glia involvement in the trigeminal system attributing to post-traumatic headache
Post-traumatic headache (PTH) is a common comorbid symptom affecting at least one-third of patients with mild traumatic brain injury (mTBI). While neuroinflammation is known to contribute to the development of PTH, the cellular mechanisms in the trigeminal system crucial for understanding the pathogenesis of PTH remain unclear. A non-invasive repetitive mTBI (4 times with a 24-hour interval) was induced in male mice and effect of mTBI was tested on either bregma or pre-bregma position on the head. Periorbital allodynia and spontaneous pain behavior were assessed using von Frey test and grimace score, respectively. Quantitative PCR was used to assess extent of mTBI pathology. RNA sequencing was performed to obtain transcriptomic profile of the trigeminal ganglion (TG), trigeminal nucleus caudalis (Sp5C) and periaqueductal gray (PAG) at 7 days post-TBI. Subsequently, quantitative PCR, in situ hybridization and immunohistochemistry were used to examine mRNA and protein expression of glia specific markers and pain associated molecules. The repetitive impacts at the bregma, but not pre-bregma site led to periorbital hypersensitivity, which was correlated with enhanced inflammatory gene expression in multiple brain regions. RNA sequencing revealed mTBI induced distinct transcriptomic profiles in the peripheral TG and central Sp5C and PAG. Using gene set enrichment analysis, positive enrichment of non-neuronal cells in the TG and neuroinflammation in the Sp5C were identified to be essential in the pathogenesis of PTH. In situ assays also revealed that gliosis of satellite glial cells in the TG and astrocytes in the Sp5C were prominent days after injury. Furthermore, immunohistochemical study revealed a close interaction between activated microglia and reactive astrocytes correlating with increased calretinin interneurons in the Sp5C. Transcriptomics analysis indicated that non-neuronal cells in peripheral TG and successive in situ assays revealed that glia in the central Sp5C are crucial in modulating headache-like symptoms. Thus, selective targeting of glia cells can be a therapeutic strategy for PTH attributed to repetitive mTBI.

CalTrig: A GUI-Based Machine Learning Approach for Decoding Neuronal Calcium Transients in Freely Moving Rodents
Advances in in vivo Ca2+ imaging using miniatured microscopes have enabled researchers to study single-neuron activity in freely moving animals. Tools such as MiniAN and CalmAn have been developed to convert Ca2+ visual signals to numerical information, collectively referred to as CalV2N. However, substantial challenges remain in analyzing the large datasets generated by CalV2N, particularly in integrating data streams, evaluating CalV2N output quality, and reliably and efficiently identifying Ca2+ transients. In this study, we introduce CalTrig, an open-source graphical user interface (GUI) tool designed to address these challenges at the post-CalV2N stage of data processing. CalTrig integrates multiple data streams, including Ca2+ imaging, neuronal footprints, Ca2+ traces, and behavioral tracking, and offers capabilities for evaluating the quality of CalV2N outputs. It enables synchronized visualization and efficient Ca2+ transient identification. We evaluated four machine learning models (i.e., GRU, LSTM, Transformer, and Local Transformer) for Ca2+ transient detection. Our results indicate that the GRU model offers the highest predictability and computational efficiency, achieving stable performance across training sessions, different animals and even among different brain regions. The integration of manual, parameter-based, and machine learning-based detection methods in CalTrig provides flexibility and accuracy for various research applications. The user-friendly interface and low computing demands of CalTrig make it accessible to neuroscientists without programming expertise. We further conclude that CalTrig enables deeper exploration of brain function, supports hypothesis generation about neuronal mechanisms, and opens new avenues for understanding neurological disorders and developing treatments.

Efficient Coding in Motor Planning
A paramount challenge for the brain is to precisely model the world and control behavior within the confines of limited encoding capacities. Efficient coding theory posits a unified framework for understanding how neural systems enhance encoding accuracy by tuning to environmental statistics. While this theory has been thoroughly explored within the perceptual realm, it is less clear how efficient coding applies to the motor system. Here, we probe the core principles of efficient coding theory through center-out reaching tasks. Our results reveal novel sequential effects in motor planning: The current movements are biased in a direction opposite to that of recent movements, and individual differences in this repulsive bias are associated with variability within the motor system. Furthermore, we observed that movement variance escalates with the angular divergence between successive movements. These findings are consistent with the prediction of an efficient coding model for motor planning and contrast with alternative models for sequential motor effects such as Bayesian models or repeated suppression.

Controllability and cause in human collaboration
There has been considerable interest in how we ascertain whether an environment is controllable and the neural mechanisms mediating this process. An especially acute version of this problem occurs when multiple people collaborate towards a joint outcome. Here we demonstrate that in such ambiguous social contexts, people engage in specific patterns of behaviour that we refer to as active disambiguation. This process helps individuals establish what they themselves, as opposed to others, control and what consequence they themselves cause or that another person causes. People identify when active disambiguation is needed and engage in it at that time. A pattern of activity in the supramarginal gyrus that emerges during and after active disambiguation is linked to establishing controllability and tracking the outcomes from which control is inferred. Activity in this brain region also signals a second learning mechanism, by which individuals attribute outcomes to themselves versus others, in proportion to their perceived control.

Separated attractors in neural landscape of motor cortex encoding motor learning
Animals gain motor learning via decrease of variation through repeated training. The secondary motor (M2) cortex shows an indispensable role in the learning process of the rotarod-learning task. Yet, it remains unclear how population decoding in M2 cortex guides the repetitive training to transform into motor enhancement. We recorded neuronal population activity using Ca2+ imaging during this enhancement revealing that neuronal population correlates of the persistent internal learning state evolves in the process of motor learning. With the behavioral micro-states analysis, we identify the growing periodicity, stability, and consistency with two gradually clearer point attractor in the M2 neural state space. The results show the evolution of attractors in M2 participate in decrease of training-acquisition behavior variation and provide a general framework for the mapping between arbitrary non-task motor learning and neural topological structure.

Spatial and Spectral Changes in Cortical Surface Potentials during Pinching versus Thumb and Index Finger Flexion
Electrocorticographic (ECoG) signals provide high-fidelity representations of sensorimotor cortex activation during contralateral hand movements. Understanding the relationship between independent and coordinated finger movements along with their corresponding ECoG signals is crucial for precise brain mapping and neural prosthetic development. We analyzed subdural ECoG signals from three adult epilepsy patients with subdural electrode arrays implanted for seizure foci identification. Patients performed a cue-based task consisting of thumb flexion, index finger flexion or a pinching movement of both fingers together. Broadband power changes were estimated using principal component analysis of the power spectrum. All patients showed significant increases in broadband power during each movement compared to rest. We created topological maps for each movement type on brain renderings and quantified spatial overlap between movement types using a resampling metric. Pinching exhibited the highest spatial overlap with index flexion, followed by superimposed index and thumb flexion, with the least overlap observed for thumb flexion alone. This analysis provides practical insights into the complex overlap of finger representations in the motor cortex during various movement types, and may help guide more nuanced approaches to brain-computer interfaces and neural prosthetics.

A comparison of microglial morphological complexity analysis in adult mouse brain samples using 2-dimensional and 3-dimensional image analysis tools
Characterizing cell morphology has been an essential aspect of neuroscience for over a century to provide essential insights into cellular function and dysfunction. Microglia, the resident innate immune cells of the central nervous system, undergo drastic changes in morphology in response to various stimuli, with many classifications proposed in recent years. Increased availability of advanced analysis software to study microglial morphology represents a step forward in the field. However, whether the use of advanced analysis tools provides equivalent or varied outcomes remains undetermined. This study re-analyzed raw data, previously processed using a standard 2D microglial morphology analysis method, using 3D analysis methods. The published article observed significant changes in microglial morphology in the mouse ventral hippocampus after administration of a ketogenic diet and exposure to repeated social defeat stress in young adult male mice. Overall, we observed different statistical outcomes in the 3D dataset compared to the previously published 2D results, with both maintained and new findings. This may indicate that the 3D analysis method is better able to capture minute changes in morphology. However, overall conclusions on microglial morphology changes remain consistent between methods. Lastly, we highlight the difference between a more mainstream statistical design and a nested design, which takes into account individual cell and animal variability. Overall, we highlight and discuss the difference between 2D and 3D microglia morphology analysis and explore the contribution of individual cell and animal variability to statistical outcomes.

High-Complexity Barcoded Rabies Virus for Scalable Circuit Mapping Using Single-Cell and Single-Nucleus Sequencing
Single cell genomics has revolutionized our understanding of neuronal cell types. However, scalable technologies for probing single-cell connectivity are lacking, and we are just beginning to understand how molecularly defined cell types are organized into functional circuits. Here, we describe a protocol to generate high-complexity barcoded rabies virus (RV) for scalable circuit mapping from tens of thousands of individual starter cells in parallel. In addition, we introduce a strategy for targeting RV-encoded barcode transcripts to the nucleus so that they can be read out using single-nucleus RNA sequencing (snRNA-seq). We apply this tool in organotypic slice cultures of the developing human cerebral cortex, which reveals the emergence of cell type-specific circuit motifs in midgestation. By leveraging the power and throughput of single cell genomics for mapping synaptic connectivity, we chart a path forward for scalable circuit mapping of molecularly-defined cell types in healthy and disease states.

Brain-wide connectivity and novelty response of the dorsal endopiriform nucleus in mice
The dorsal endopiriform nucleus (EPd) is an enigmatic cortical subplate structure located inside the piriform cortex that shares a similar developmental origin with the claustrum. Although the EPd has been previously implicated in epilepsy and olfactory processing, its anatomical organization, connectivity patterns, and function remain largely unclear due to a lack of specific molecular markers. Our previous mapping study serendipitously identified that the Oxt receptor (Oxtr) is densely expressed in the EPd. Subsequent immunohistochemical and spatial transcriptomic analyses confirmed that Oxtr expression is enriched in the EPd, revealing distinct molecular organization compared to the neighboring claustrum. Whole brain input-output mapping of EPd Oxtr-positive neurons unveils extensive bidirectional connections to the ventral half of the brain, orchestrating functional circuits regulating olfaction, internal state, and emotion. Furthermore, our in-vivo miniscope recordings show that EPd Oxtr neurons exhibit high baseline activity during exploratory behavior, with a sharp decrease in activity in response to novel stimuli. This suggests that the EPd regulates interoceptive states and likely plays a role in adapting to novel exteroceptive cues.

Experience shapes the transformation of olfactory representations along the cortico-hippocampal pathway
Perception relies on the neural representation of sensory stimuli. Primary sensory cortical representations have been extensively studied, but how sensory information propagates to memory-related multisensory areas has not been well described. We studied this question in the olfactory cortico-hippocampal pathway in mice. We recorded single units in the anterior olfactory nucleus (AON), the anterior piriform cortex (aPCx), lateral entorhinal cortex (LEC), the hippocampal CA1 subfield, and the subiculum (SUB) while animals performed a non-associative learning paradigm involving novel and familiar stimuli. Novel stimuli evoked larger responses than familiar stimuli in the AON, whereas in hippocampal areas, novelty was reflected by the number of responsive neurons. In parallel, odorant selectivity increased along the pathway. While both stimulus identity and experience were thus reflected in all regions, their neural representations gradually separated. Our findings provide a potential mechanism for how sensory representations are transformed to support stimulus identification and implicit memories.

Reduced Synaptic Heterogeneity in a Tetanus ToxinModel of Epilepsy: Insights from ComputationalModeling
A neural mass model was used to assess connectivity strength across diverse populations by fitting the model to background EEG data obtained from a Tetanus Toxin rat model of epilepsy. Our findings reveal a notable decline in the variability of estimated parameters when using EEG data recorded from rats in the Tetanus Toxin group compared with the control group. A detailed comparison of standard deviations in estimated parameters between day 1 and day 20 recordings, coinciding with a heightened number of seizures, underscores the impact of Tetanus Toxin on diminishing synaptic strength variability across recordings. This study supports electrophysiological studies suggesting that epileptogenesis induces a reduction in biophysical heterogeneity, potentially leading to an increase in network synchrony associated with epilepsy. Furthermore, our computational model establishes a foundation for future explorations of the implications of this diminished variability.

Age dependency of neurometabolite T1 relaxation times
Purpose To measure T1 relaxation times of metabolites at 3T in a healthy aging population and investigate age dependence. Methods A cohort of 101 healthy adults were recruited with approximately 10 male and 10 female participants in each decade band: 18-29, 30-39, 40-49, 50-59, and 60+ years old. Inversion-recovery PRESS data (TE/TR: 30/2000 ms) were acquired at 8 inversion times (TIs) (300, 400, 511, 637, 780, 947, 1148 and 1400 ms) from voxels in white-matter-rich centrum semiovale (CSO) and gray-matter-rich posterior cingulate cortex (PCC). Modeling of TI-series spectra was performed in Osprey 2.5.0. Quantified metabolite amplitudes for total N-acetylaspartate (tNAA2.0), total creatine at 3.0 ppm (tCr3.0) and 3.9 ppm (tCr3.9), total choline (tCho), myo-inositol (mI), and the sum of glutamine and glutamate (Glx) were modeled to calculate T1 relaxation times of metabolites. Results T1 relaxation times of tNAA2.0 in CSO and tNAA2.0, tCr3.0, mI and Glx in PCC decreased with age. These correlations remained significant when controlling for cortical atrophy. T1 relaxation times were significantly different between PCC and CSO for all metabolites except tCr3.0. We also propose linear models for predicting metabolite T1s at 3T to be used in future aging studies. Conclusion Metabolite T1 relaxation times change significantly with age, an effect that will be important to consider for accurate quantitative MRS, particularly in studies of aging.

Remote activation of place codes by gaze in a highly visual animal
Vision enables many animals to perform spatial reasoning from remote locations. By viewing distant landmarks, animals recall spatial memories and plan future trajectories. Although these spatial functions depend on hippocampal place cells, the relationship between place cells and active visual behavior is unknown. Here, we studied a highly visual animal, the chickadee, in a behavior that required alternating between remote visual search and spatial navigation. We leveraged the unique head-directed nature of avian vision to track gaze in freely moving animals. We discovered a profound link between place coding and gaze. Place cells activated not only when the chickadee was in a specific location, but also when it simply gazed at that location from a distance. Gaze coding was precisely timed by fast, ballistic head movements called "head saccades". On each saccadic cycle, the hippocampus switched between encoding a prediction of what the bird was about to see and a reaction to what it actually saw. The temporal structure of these responses was coordinated by subclasses of interneurons that fired at different phases of the saccade. We suggest that place and gaze coding are components of a unified process by which the hippocampus represents the location that an animal is currently attending to. This process allows the hippocampus to implement both local and remote spatial functions.

Autophagic enhancer rescues Tau accumulation in a stem cell model of frontotemporal dementia
Tau degradation is disrupted in neurodegenerative tauopathies, such as frontotemporal dementia (FTD), which may contribute to Tau aggregation. The prevailing hypothesis has been that Tau degradation is stymied due to an imbalance in proteostasis that occurs with age. Here, we used Airyscan super resolution imaging to illustrate that a pathogenic FTD mutation in the MAPT gene, which encodes Tau, is sufficient to alter multiple steps of the autophagy lysosomal pathway and impair Tau degradation. We discovered lysosomes clogged with both Tau and phosphorylated Tau, stalled lysosome motility, disrupted molecular motors, enhanced autophagic flux, and slowed cargo degradation in mutant Tau neurons. Treatment of mutant Tau neurons with a small molecule autophagy enhancer drug increases autophagic flux and cargo degradation, reduces phospho-Tau levels, and reduces Tau accumulation in lysosomes without restoring defects in lysosomal motility. This study reveals novel effects of mutant Tau and provides a window through which therapeutic treatments targeting autophagy may promote Tau homeostasis.

Circulatory proteins shape microglia state and boost phagocytosis
Microglia, the brain immune cells, are highly responsive to their local environment. Given that circulatory proteins can enter the brain, we asked whether microglia are responsive to such proteins. Here, we identify a stable population of microglia specialized to take up circulatory proteins in a region-specific manner under physiological conditions; human hematopoietic stem cell-derived microglia replacing endogenous microglia in chimeric mice show similar regional specialization. Plasma-positive microglia are characterized by prominent expression of genes related to innate immunity and antigen presentation and exhibit high metabolic and phagocytic activity. This activity is dependent, in part, on microglial uptake and accumulation of circulatory Apolipoprotein AI (ApoA-I). Our findings thus identify a new model of communication between brain and periphery through specialized microglia.

Loss of SARM1 protects against retinal ganglion cell degeneration in Autosomal Dominant Optic Atrophy
Autosomal Dominant Optic Atrophy (ADOA), the most prevalent inherited optic neuropathy, leads to retinal ganglion cell (RGC) degeneration and vision loss. ADOA is primarily caused by mutations in the OPA1 gene, which encodes a conserved GTPase important for mitochondrial inner membrane dynamics. To date, the disease mechanism remains unclear, and no therapies are available. Here, we present a novel mouse model carrying the pathogenic Opa1R290Q/+ allele that recapitulates key features of human ADOA, including mitochondrial defects, age-related RGC loss, optic nerve degeneration, and reduced RGC functions. We identify SARM1, a neurodegeneration switch, as a key driver of RGC degeneration in these mice. Sarm1 knockout nearly completely suppresses all the degeneration phenotypes. Additionally, we show that SARM1 is located within the mitochondrial intermembrane space (IMS). These findings indicate that SARM1 is activated downstream of mitochondrial dysfunction in ADOA, highlighting it as a promising therapeutic target.

Neuronal loss of Galnt2 Impairs O-glycosylation and Leads to Neurobehavioral Deficits Mimicking GALNT2-CDG
GALNT2-CDG is a multi-system genetic disorder due to biallelic pathogenic mutations in GALNT2, which encodes a ubiquitously expressed Golgi-localized glycosyltransferase that initiates mucin-type O-glycosylation. Affected individuals exhibit dysmorphic facial features, short stature, decreased HDL-C, and notable impairments in brain function. GALNT2-CDG patients show global developmental delay without speech development, childhood epilepsy, autistic-like features, and white-matter brain abnormalities. The extent of O-glycosylation in brain development and function remains poorly understood. To address this question, we selectively ablated Galnt2 from pan-neuronal cells in the brain and found that conditional knockout mice exhibit deficits across numerous behavioral domains, including locomotion, motor coordination, sociability, learning, and memory, as well as experience spontaneous seizures, recapitulating characteristic neurological manifestations of GALNT2-CDG. Given the catalytic activity of GALNT2 to initiate mucin-type O-glycosylation, we used glycoproteomics to identify disrupted O-glycosylation in synaptosomes purified from cortical tissues. We ascertained a non-redundant, isoform-specific contribution of GALNT2 to the cortical synaptosomal O-glycoproteome, identifying candidate glycoproteins and disrupted O-glycosites that accompany behavioral abnormalities in knockout mice. These findings demonstrate functional impact of O-glycosylation in neurons, implicating roles of O-glycosylation in diverse molecular and cellular pathways related to neuronal function and provide new opportunities to gain insights into the neurological pathophysiology of GALNT2-CDG.

Heartbeat related activity in the anterior thalamus differs between phasic and tonic REM sleep periods
Rapid eye movement (REM) sleep is a fundamental sleep state associated with diverse functions from elemental physiological processes to higher order neurocognitive functions. A growing body of research indicates that REM sleep with eye movements (phasic REM) differs from REM periods without ocular activity (tonic) in terms of spontaneous and evoked neural responses. Studies using auditory stimulation consistently observed enhanced evoked responses in tonic versus phasic REM, indicating that the external processing is largely diminished when the eyes move during REM sleep. Whereas exteroceptive processing during sleep is widely studies, investigation on interoception (the processing of bodily signals) during sleep is scarce, and limited to scalp electroencephalographic recordings. Here we studied interoceptive processing in a group of epileptic patients (N = 11) by measuring their heartbeat-related neural activity in the anterior nucleus of the thalamus (ANT) during phasic and tonic REM sleep and resting wakefulness. Evoked potentials and beta-low gamma spectral power locked to the heartbeat were significantly different in phasic REM compared to tonic REM and wakefulness. Heartbeat-related neural signals exhibited pronounced inter-trial phase synchronization at lower (7-20 Hz) oscillatory activity in all vigilance states, but reduced gamma synchronization at later time points in phasic REM only. Tonic REM and wakefulness did not show significant differences in heartbeat-related activity in the ANT. Our findings indicate that heartbeat-related neural activity is detectable at the level of the ANT, showing distinct signatures of interoceptive processing in phasic REM compared to tonic REM and wakefulness.

Posterior Parietal Cortex Regulates Intrinsic Neural Timescales and Attentional Processing in Frontal Eye Field
Intrinsic neural timescales characterize the dynamics of endogenous fluctuations in neural activity. We measured the intrinsic timescales of frontal eye field (FEF) neurons and examined changes during posterior parietal cortex (PPC) inactivation. FEF neurons exhibit a bimodal distribution of intrinsic timescales, with shorter timescale neurons processing rapid visual information and longer timescale neurons more involved in sustained attentional modulation. PPC inactivation significantly increased intrinsic timescales in both neuron types, with a 15-fold greater increase in shorter timescale neurons. Additionally, PPC inactivation reduced visual and attentional responses, with a stronger effect on attention in longer timescale neurons. This disruption eliminated the correlation between timescales and attentional responses observed in the control condition. Our results provide the first causal evidence that FEF intrinsic timescales depend on long-range projections from PPC, suggesting the presence of at least two network motifs with different timescales that contribute to neuronal dynamics and functional computations within FEF.