bioRxiv Subject Collection: Neuroscience's Journal
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Friday, January 24th, 2025
| Time |
Event |
| 12:19a |
Erythropoietin decreases apoptosis and promotes Schwann cell repair and phagocytosis following nerve crush injury in mice
After peripheral nerve trauma, insufficient clearance of phagocytic debris significantly hinders nerve regeneration. Without sufficient myelin debris clearance, Schwann cells (SCs) undergo increased apoptosis, impairing functional recovery. There is no treatment for peripheral nerve crush injury (PNCI). Erythropoietin (EPO) is an FDA-approved drug for anemia, which may help in the treatment of PNCI by transdifferentiating resident SCs into repair SCs (rSCs) and enhancing phagocytosis to facilitate the removal of cellular debris. For the first time, we conducted bulk RNA sequencing on mice with calibrated sciatic nerve crush injuries (SNCIs) on days 3, 5, and 7 post-SNCI to uncover transcriptomic changes with and without EPO treatment. We found EPO altered several biological pathways and associated genes, particularly those involved in cell apoptosis, differentiation, proliferation, phagocytosis, myelination, and neurogenesis. We validated the effects of EPO on SNCI on early (days 3/5) and intermediate (day 7) post-SNCI, and found EPO treatment reduced apoptosis (TUNEL), and enhanced SC repair (c-Jun and p75-NTR), proliferation (Ki67), and the phagocytosis of myelin debris by rSCs at crush injury sites. This improvement corresponded with an enhanced sciatic functional index (SFI). We also confirmed these findings in-vitro. EPO significantly enhanced SC repair during early de-differentiation, marked by high c-Jun and p75-NTR protein levels, and later re-differentiation with high EGR2 and low c-Jun and p75-NTR levels. These changes occurred under lipopolysaccharide (LPS) stress at 24 and 72h, respectively, compared to LPS treatment alone. Under LPS stress, EPO also significantly increased rSCs proliferation and phagocytosis of myelin or dead SCs. In conclusion, our findings support EPO may enhance the function of rSCs in debris clearance as a basis for its possible use in treating nerve trauma. | | 12:19a |
Homeostatic bidirectional plasticity in Upbound and Downbound micromodules of the olivocerebellar system
Olivocerebellar learning is highly adaptable, unfolding over minutes to weeks depending on the task. However, the stabilizing mechanisms of the synaptic dynamics necessary for ongoing learning remain unclear. We constructed a model to examine plasticity dynamics under stochastic input and investigate the impact of inferior olive (IO) reverberations on Purkinje cell (PCs) activity and synaptic plasticity. We explored Upbound and Downbound cerebellar micromodules, which are organized loops of IO neurons, cerebellar nuclei neurons and microzones of PCs characterized by their unique molecular profiles and different levels of baseline firing. Our findings show synaptic weight convergence followed by stability of synaptic weights. In line with their relatively low and high intrinsic firing, we observed that Upbound and Downbound PCs have a propensity for potentiation and depression, respectively, with both PC types reaching stability at differential levels of overall strength of their parallel-fiber (PF) inputs. The oscillations and coupling of IO neurons participating in the Upbound and Downbound modules determine at which frequency band PFs can be stabilized optimally. Our results indicate that specific frequency components drive IO resonance and synchronicity, which, in turn, regulate temporal patterning across Upbound and Downbound zones, orchestrating their plasticity dynamics. | | 12:19a |
Deficient Executive Control in Transformer Attention
Although transformers in the large language models (LLMs) effectively implement a self-attention mechanism that has revolutionized natural language processing, they lack an explicit implementation of executive control of attention found in humans which is essential for resolving conflicts and selecting relevant information in the presence of competing stimuli, and is critical for adaptive behavior. To investigate this limitation in LLMs, we employed the classic color Stroop task that is widely regarded as the gold standard for testing executive control of attention. Our results revealed a typical conflict effect of better performance in terms of accuracy in the congruent condition (e.g., naming the ink color of the word RED) compared to incongruent condition (e.g., naming the ink color of the word RED), which is similar to human performance, in short sequences. However, as sequence length increased, the performance degraded toward chance levels on the incongruent trials despite maintaining excellent performance on congruent trials and near-perfect word reading ability. These findings demonstrate that while transformer attention mechanisms can achieve human-comparable performance in smaller contexts, they are fundamentally limited in their capacity for conflict resolution across extended contexts. This study suggests that incorporating executive control mechanisms akin to those in biological attention could be crucial for achieving more general reasoning and reliable performance toward artificial general intelligence. | | 12:19a |
Resolving inconsistent effects of tDCS on learning using a homeostatic structural plasticity model
Transcranial direct current stimulation (tDCS) is increasingly used to modulate motor learning. Current polarity and intensity, electrode montage, and application before or during learning had mixed effects. Both Hebbian and homeostatic plasticity were proposed to account for the observed effects, but the explanatory power of these models is limited. In a previous modeling study, we showed that homeostatic structural plasticity (HSP) can explain long-lasting after-effects of tDCS and transcranial magnetic stimulation (TMS). The interference between motor learning and tDCS, which are both based on HSP in our model, is a candidate mechanism to resolve complex and seemingly contradictory experimental observations. We implemented motor learning and tDCS in a spiking neural network subject to HSP. The anatomical connectivity of the engram induced by motor learning was used to quantify the impact of tDCS on motor learning. We demonstrated with modeling that transcranial direct current stimulation applied before learning had weak modulatory effects. It led to a small reduction in connectivity if it was applied uniformly. When applied during learning, targeted anodal stimulation significantly strengthened the engram, while targeted cathodal or uniform stimulation weakened it. Applied after learning, targeted cathodal, but not anodal, tDCS boosted engram connectivity. Strong tDCS would distort the engram structure if not applied in a targeted manner. Our model explained both Hebbian and homeostatic phenomena observed in human tDCS experiments by assuming memory strength positively correlates with engram connectivity. This includes applications with different polarity, intensity, electrode montage, and timing relative to motor learning. The HSP model provides a promising framework for unraveling the dynamic interaction between learning and transcranial DC stimulation. | | 12:19a |
Co-Conservation of Synaptic Gene Expression and Circuitry in Collicular Neurons
The superior colliculus (SC), a midbrain sensorimotor hub, is anatomically and functionally similar across vertebrates, but how its cell types have evolved is unclear. Using single-nucleus transcriptomics, we compared the SC molecular and cellular organization in mice, tree shrews, and humans. Despite over 96 million years of evolutionary divergence, we identified ~30 consensus neuronal subtypes, including Cbln2+ neurons that form the SC-pulvinar circuit in mice and tree shrews. Synapse-related genes were among the most conserved, unlike neocortex, suggesting co-conservation of synaptic genes and circuitry. In contrast, cilia-related genes diverged significantly across species, highlighting the potential importance of the neuronal primary cilium in SC evolution. Additionally, we identified a novel inhibitory SC neuron in tree shrews and humans but not mice. Our findings reveal that the SC has evolved by conserving neuron subtypes, synaptic genes, and circuitry, while diversifying ciliary gene expression and an inhibitory neuron subtype. | | 12:19a |
Chronic chemogenetic slow-wave sleep enhancement in mice.
While epidemiological associations and brief studies of sleep effects in human disease have been conducted, rigorous long-term studies of sleep manipulations and in animal models are needed to establish causation and to understand mechanisms. We have previously developed a mouse model of acute slow-wave-sleep (SWS) enhancement using chemogenetic activation of parafacial zone GABAergic neurons (PZGABA) in the parvicellular reticular formation of the pontine brainstem. However, it was unknown if SWS could be enhanced chronically in this model. In the present study, mice expressing the chemogenetic receptor hM3Dq in PZGABA were administered daily with one of three chemogenetic ligands, clozapine N-oxide (CNO), deschloroclozapine (DCZ) and compound 21 (C21), and sleep-wake phenotypes were analyzed using electroencephalogram (EEG) and electromyogram (EMG). We found that SWS time is increased for three hours, and at the same magnitude for at least six months. This phenotype is associated with an increase of slow wave activity (SWA) of similar magnitude throughout the 6-month dosing period. Interestingly, at the end of the 6-month dosing period, SWA remains increased for at least a week. This study validates a mouse model of chronic SWS enhancement that will allow mechanistic investigations into how SWS promotes physiological function and prevents diseases. The approach of a rotating schedule of three chemogenetic ligands may be broadly applicable in chemogenetic studies that require chronic administration. | | 12:19a |
Functional recovery of adult brain tissue arrested in time during cryopreservation by vitrification
Cryopreserving adult brain tissue is challenging due to damage from ice formation, and traditional freezing methods fail to maintain neural architecture and function. Vitrification offers a promising alternative but has not been surveyed in the brain. Here, we demonstrate near-physiological recovery of adult mouse hippocampal slices after vitrification. Key features of this cognitive hub are preserved, including structural integrity, metabolic responsiveness, neuronal excitability, and synaptic transmission and plasticity. Notably, hippocampal long-term potentiation was well preserved, indicating that the cellular machinery of learning and memory remains operational. These findings extend the knowledge of brain recovery after hypothermic shutdown to recovery after complete cessation of molecular mobility through vitrification. This suggests that brain tissue can be arrested in time and reactivated, opening avenues for potential clinical applications. | | 4:40a |
Proximity labeling and orthogonal nanobody pulldown (ID-oPD) approaches to map the spinophilin interactome uncover a putative role for spinophilin in protein homeostasis.
Spinophilin is a dendritic spine enriched scaffolding and protein phosphatase 1 targeting protein. To detail spinophilin interacting proteins, we created an Ultra-ID and ALFA-tagged spinophilin encoding construct that permits proximity labeling and orthogonal nanobody pulldown (ID-oPD) of spinophilin-associated protein complexes in heterologous cells. We identified 614 specific, and 312 specific and selective, spinophilin interacting proteins in HEK293 cells and validated a subset of these using orthogonal approaches. Many of these proteins are involved in mRNA processing and translation. In the brain, we determined that spinophilin mRNA is highly neuropil localized and that spinophilin may normally function to limit its own expression but promote the expression of other PSD-associated proteins. Overall, our use of an ID-oPD approach uncovers a novel putative role for spinophilin in mRNA translation and synaptic protein expression specifically within dendritic spines. | | 4:40a |
An Open-Source Joystick Platform for Investigating Forelimb Motor Control, Auditory-Motor Integration, and Value-Based Decision-Making in Head-Fixed Mice
Investigation of neural processes underlying motor control requires behavioral readouts that capture the richness of actions, including both categorical (choice-based) information and motor execution (kinematics). We present an open-source platform for behavioral training of head-fixed mice that combines a stationary or retractable forelimb-based joystick, sound-presentation system, capacitive lick sensor, and water reward dispenser. The setup allows for the creation of multiple behavioral paradigms, two of which are highlighted here: a two-alternative forced-choice auditory-motor discrimination paradigm, and a two-armed bandit value-based decision-making task. In the auditory-motor paradigm, mice learn to report high or low frequency tones by pushing or pulling the joystick. In the value-based paradigm, mice learn to push or pull the joystick based on the history of rewarded trials. In addition to reporting categorical choices, this setup provides a rich dataset of motor parameters that reflect components of the underlying learning and decision processes in both of these tasks. These kinematic parameters (including joystick speed and displacement, Frechet similarity of trajectories, tortuosity, angular standard deviation, and movement vigor) provide key additional insights into the motor execution of choices that are not as readily assessed in other paradigms. The systems flexibility of task design, joystick readout, and ease of construction represent an advance compared to currently available manipulandum tasks in mice. We provide detailed schematics for constructing the setup and protocols for behavioral training using both paradigms, with the hope that this open-source resource is readily adopted by neuroscientists interested in mechanisms of sensorimotor integration, motor control, and choice behavior. | | 4:40a |
Data-driven denoising in spinal cord fMRI with principal component analysis
Numerous approaches have been used to denoise spinal cord functional magnetic resonance imaging (fMRI) data. Principal component analysis (PCA)-based techniques, which derive regressors from a noise region of interest (ROI), have been used in both brain (e.g., CompCor) and spinal cord fMRI. However, spinal cord fMRI denoising methods have yet to be systematically evaluated. Here, we formalize and evaluate a PCA-based technique for deriving nuisance regressors for spinal cord fMRI analysis (SpinalCompCor). In this method, regressors are derived with PCA from a noise ROI, an area defined outside of the spinal cord and cerebrospinal fluid. A parallel analysis is used to systematically determine how many components to retain as regressors for modeling; this designated a median of 11 regressors across three fMRI datasets: motor task (n=26), breathing task (n=27), and resting state (n=10). First-level fMRI modeling demonstrated that principal component regressors did fit noise (e.g., physiological noise from blood vessels), particularly in the resting state fMRI dataset. However, group-level motor task activation maps themselves did not show a clear benefit from including SpinalCompCor regressors over our original denoising model. The potential for collinearity of principal component regressors with the task may be a concern, and this should be considered in future implementations for which task-correlated noise is anticipated. | | 4:40a |
Evaluation of distal facial nerve branches contribution to facial nerve paralysis in rodents
Introduction/Aims: Facial nerve paralysis is a complex and devastating condition. Translational research of facial paralysis recovery remains largely limited to animal studies, for which there are many potential models employed. When studying facial nerve regeneration in rodents, it is important to understand the converging contributions of the motor supply into the whisker pad. A consensus surgical approach and animal model has yet to be defined. Of particular interest for movement of the nose and whiskers are the buccal and marginal mandibular nerves. This study aims to evaluate how these distal nerve branches contribute to facial nerve paralysis and identify key morphological changes at the neuromuscular junctions (NMJs) in the whisker pad of rodents. Methods: Adult rats underwent isolated transection of the buccal branch of the facial nerve, both the buccal and marginal mandibular branches of the facial nerve, or control sham surgery. Results: Histological, electrophysiological, and behavior assessments confirmed that the transection of the buccal branch alone did not cease whisker movement in rats, but when combined with a transection of the marginal mandibular branch, it resulted in full paralysis of the whisker and nose movement. Discussion: These results are indicative of the distinct roles of these nerves branches in facial paralysis repair following a transection injury. Further, our results suggest additional targets for facial nerve repair treatments. | | 4:40a |
Maturation of Hippocampus-Medial Prefrontal Cortex Projections Defines a Pathway-Specific Sensitive Period for Cognitive Flexibility
The septotemporal axis of the hippocampus separates it into domains with unique molecular, cellular, downstream connectivity and behavioral profiles, and yet very little is known about the ontogenesis of these highly specialized subcircuits. Here, we used viral tracing, optogenetic-assisted patch clamping, chemogenetics and behavior in mice to examine changes in domain-defined hippocampus efferent projections from postnatal day (P)10 to P60. We found distinct anatomical and synaptic developmental signatures in ventral and intermediate CA1 downstream connectivity, with unique contributions to the prelimbic and infralimbic subregions of the medial prefrontal cortex (mPFC). Inhibition of the ventral CA1 (vCA1)-mPFC pathway during juvenility led to a deficit in adult cognitive flexibility exclusively in females, establishing a sex- and pathway-specific sensitive period preceding the stabilization of vCA1-mPFC synaptic transmission. Our data elucidate domain- and target-defined postnatal maturation of hippocampus efferents, identifying a sex-specific sensitive period with crucial implications for early life influences on adult cognition. | | 4:40a |
Hypothalamic Vasopressin Neurons Enable Maternal Thermoregulatory Behaviors
Newborns of many mammalian species are partial poikilotherms and require adult thermoregulatory care for survival. In mice, pup survival in cold and cool ambient temperature depends on the ability of adult caregivers to huddle pups and bring them into a high-quality nest. It is therefore essential that adult mice adjust parental care as a function of changes in ambient temperature. Here, we investigated how mouse maternal care adapts to a range of temperatures, from cold to warm. We show that changes in ambient temperature affect several individual and co-parenting maternal behaviors in both dams and virgin female mice, and modulate activity of vasopressin neurons. Furthermore, we establish that the effects of ambient temperature on both maternal care and the activity of vasopressin neurons depend in part on thermosensation, specifically on the TRPM8 sensor. Using trans-synaptic anterograde tracing and whole-brain activity mapping, we find that vasopressin neurons from the paraventricular hypothalamic nucleus connect synaptically with temperature-responsive brain structures implicated in maternal care. We then show that optogenetic activation of vasopressin projections to the central amygdala, a structure activated by cold ambient temperature, recapitulates the effects of cold on co-parenting behaviors. Our data provide a biological mechanism for maternal thermoregulatory behavior in mice with translational relevance to the reported association between ecosystem temperature fluctuations and variations in human child neglect cases. | | 4:40a |
Exploring GPCR-mediated optogenetic modulation of seizure network in a pig model of Temporal Lobe Epilepsy
Rationale: Optogenetics offers unmatched cellular specificity and control over cellular activity. Various opsins have been tested in animal models of epilepsy, each contributing to our understanding of seizure circuit dynamics. However, inhibitory optogenetic tools based on microbial rhodopsins have low light sensitivity and, thus, are less suitable for applications involving larger brains. We evaluated eOPN3, a red-shifted, highly sensitive inhibitory G- protein coupled receptor opsin in a porcine seizure model using integrated electro-optical sensing and modulation. The results demonstrated the feasibility of eOPN3 circuit modulation in a large animal epilepsy model. Methods: MRI-guided stereotactic surgery was used to deliver 20-60 L of AAV_eOPN3 (AAV5/AAV9-CaMKII-eOPN3-mScarlet) into the hippocampus (HPC) of three Goottingen minipigs. Each hemisphere received either an active or a control viral vector (AAV5/9-CaMKII-mScarlet) with gadolinium to visualize the injection sites and diffusion volume via post-operative MRI. Two to three months post-injection, bilateral deep brain stimulation electrodes integrated with optic fibers were stereotactically implanted into the anterior nucleus of the thalamus (ANT) and HPC to assess: 1) opsin expression using fiber photometry, 2) optogenetic modulation of stimulation evoked response potentials (SERPs), 3) induction and propagation of seizure-like activity via intrahippocampal kainic acid (KA) injection, and 4) optogenetic modulation of KA-induced seizure activity. After the electrophysiology recording, brains were harvested for histological analysis to evaluate injection target precision, eOPN3 expression, and estimate eOPN3-modulated volume. Results: eOPN3 expression was confirmed during surgery via fiber photometry. ANT electrical stimulation elicited robust SERPs in the HPCs, which were attenuated by HPC light illumination. HPC stimulation similarly induced SERPs in the ipsilateral ANT and the contralateral HPC. The HPC stimulation-induced SERPs were significantly reduced by illuminating the site of the recording areas, the ipsilateral ANT and the contralateral HPC, demonstrating the optogenetic inhibition of the synaptic release from the HPC. KA injection into the HPC induced 20-30 Hz seizure-like activity. The ANT and HPC light illumination suppressed the localized KA-induced seizure activity in the early stage. However, after the generalization of KA-induced seizures, the ANT-HPC illumination lost efficacy for the control of seizures. Histological analysis confirmed eOPN3 expression in the HPC, ANT and other Papez circuit nodes. Conclusion: Our pilot study highlights that eOPN3-mediated inhibition alters SERP and the latency and spread of KA-induced seizure-like activity. We developed a platform incorporating pre- and postoperative MRI for precise viral vector delivery, real-time fiber photometry for quantifying opsin expression, and integrated electro-optical sensing and stimulation to assess optogenetic efficacy in a large animal model. The large animal model provides a solid foundation for future translational research to develop electro-optical devices and cellular therapies for human epilepsy. | | 8:32a |
The brain dynamics of congenitally blind people seeing faces with sound
Sensory substitution devices (SSDs) convert images to sounds to equip blind individuals with nominally visual functions, like face or letter sensitivity. Prior studies showed that image-to-sound SSDs engage cortices ordinarily specialised for visual functions. However, the brain dynamics of SSD-supported perception remains unknown. Either visual cortices are the first locus of discrimination of SSD percepts, or their activation is a by-product of perceptual processes unfurling elsewhere, such as in auditory cortices. Resolving this uncertainty is critical for understanding whether the blind truly "see" via SSDs. Using electrical neuroimaging of EEG data from congenitally blind adults, we show for the first time that inferotemporal visual cortices are the earliest site of face sensitivity when conveyed via SSDs. The blind do indeed "see" with SSDs. By providing the temporal dynamics of SSD perception, our findings provide unique evidence for the theory that cortices are characterised by task-contingent functional organisation. | | 8:32a |
Frontal midline theta power accounts for inter-individual differences in motor learning ability
Recent neurophysiological studies have demonstrated that frontal midline theta (FMT) activity plays a significant role in motor learning. One of the key challenges in motor learning is to understand the interindividual variability in learning proficiency rates, yet the underlying neural mechanisms remain unclear. To address this open question, this study recorded electroencephalogram activity during a visuomotor tracking task to investigate whether modulation of FMT power and the consistency of theta phase during motor preparation could explain individual differences in learning proficiency. We found a significant positive correlation between increased FMT power during motor preparation and learning proficiency rates. Specifically, individuals with greater FMT power exhibited faster learning rates. In contrast to this, no significant correlation was observed between the consistency of theta phase during motor preparation and learning proficiency. Together, these findings highlight that the FMT power, rather than phase synchrony, is closely associated with motor learning efficiency. This study provides a novel perspective for understanding the causes of individual differences in motor learning and further corroborates the previous evidence showing FMT power contributes to motor learning processes. |
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