bioRxiv Subject Collection: Neuroscience's Journal

Characterization of Eye and Adnexal Tissues in Dogs and Wolves: A Histological and Lectin-Histochemical Approach
This study explores the ocular anatomy and glandular components of domestic dogs compared to their ancestor, the wolf, with the aim of identifying evolutionary changes due to domestication and their implications for ocular pathologies. Utilizing histological and histochemical techniques, including hematoxylin-eosin, PAS, Alcian Blue, and lectins, this research conducts a detailed analysis of the canine and wolf ocular systems, particularly focusing on the eyelids, tarsal glands, and conjunctival tissues. Results indicate significant histological differences between the two species, particularly in the thickness and secretion levels of the conjunctival epithelia and the structure of the tarsal glands. Dogs exhibit a thicker epithelium with greater PAS and Alcian Blue positive secretion, suggesting enhanced ocular protection and lubrication adapted to domestic environments. Conversely, wolves display more concentrated glandular secretions and a predominance of acidic mucopolysaccharides, aligning with their adaptation to natural habitats. This study also highlights the translational value of dogs as models for human ocular diseases, given their anatomical and physiological similarities with humans. Such comparisons are essential as they provide insights that can lead to advancements in medical research and clinical applications, especially in the development of treatments for ocular surface disorders.

Spatial dynamics of mammalian brain development and neuroinflammation by multimodal tri-omics mapping
The ability to spatially map multiple layers of the omics information over different time points allows for exploring the mechanisms driving brain development, differentiation, arealization, and alterations in disease. Herein we developed and applied spatial tri-omic sequencing technologies, DBiT ARP-seq (spatial ATAC-RNA-Protein-seq) and DBiT CTRP-seq (spatial CUT&Tag-RNA-Protein-seq) together with multiplexed immunofluorescence imaging (CODEX) to map spatial dynamic remodeling in brain development and neuroinflammation. A spatiotemporal tri-omic atlas of the mouse brain was obtained at different stages from postnatal day P0 to P21, and compared to the regions of interest in the human developing brains. Specifically, in the cortical area, we discovered temporal persistence and spatial spreading of chromatin accessibility for the layer-defining transcription factors. In corpus callosum, we observed dynamic chromatin priming of myelin genes across the subregions. Together, it suggests a role for layer specific projection neurons to coordinate axonogenesis and myelination. We further mapped the brain of a lysolecithin (LPC) neuroinflammation mouse model and observed common molecular programs in development and neuroinflammation. Microglia, exhibiting both conserved and distinct programs for inflammation and resolution, are transiently activated not only at the core of the LPC lesion, but also at distal locations presumably through neuronal circuitry. Thus, this work unveiled common and differential mechanisms in brain development and neuroinflammation, resulting in a valuable data resource to investigate brain development, function and disease.

Single-nucleus and spatial transcriptomic analysis identified molecular features of neuronal heterogeneity and distinct glial responses in Parkinson's disease.
The heterogeneity of Parkinson's disease (PD) is increasingly recognized as an important aspect of understanding the disorder. Among the factors contributing to this heterogeneity, ethnic differences are primary sources, significantly influencing the likelihood of PD developing and its initial symptoms' nature. While there have been numerous reports related to PD in East Asia, there has been a lack of contribution from single-cell (or nucleus) transcriptome studies, which have been making significant contributions to understanding PD. In this study, a total of 33,293 nuclei obtained from the substantia nigra (SN) of confirmed pathological PD and control patients in South Korea were profiled, revealing eight different cell types through cluster analysis. Monocle-based pseudotime analysis identified two disease-associated trajectories for each astrocyte and microglia and identified genes that differentiate them. Interestingly, we uncovered the inflammatory intervention in the early PD-associated transition in microglia and identified the molecular features of this intermediate state of microglia. In addition, gene regulatory networks (GRNs) based on TENET analysis revealed the detrimental effect of an HSPA5-led module in microglia and MSRB3- and HDAC8- led modules specifying the two different astrocyte trajectories. In SN neurons, we observed population changes, a decrease in dopaminergic and glutamatergic neurons and a proportional increase in GABAergic neurons. By deconvolution in spatial transcriptome obtained the PD sample, we confirmed spatiotemporal heterogeneity of neuronal subpopulations and PD-associated progressive gliosis specific to dopaminergic nuclei, SN and ventral tegmental areas (VTAs). In conclusion, our approach has enabled us to identify the genetic and spatial characterization of neurons and to demonstrate different glial fates in PD. These findings advance our molecular understanding of cell type-specific changes in the progression of Korean PD, providing an important foundation for predicting and validating interventions or drug effects for future treatments.

Context-dependence of deterministic and nondeterministic contributions to closed-loop steering control
In natural circumstances, sensory systems operate in a closed loop with motor output, whereby actions shape subsequent sensory experiences. A prime example of this is the sensorimotor processing required to align one's direction of travel, or heading, with one's goal, a behavior we refer to as steering. In steering, motor outputs work to eliminate errors between the direction of heading and the goal, modifying subsequent errors in the process. The closed-loop nature of the behavior makes it challenging to determine how deterministic and nondeterministic processes contribute to behavior. We overcome this by applying a nonparametric, linear kernel-based analysis to behavioral data of monkeys steering through a virtual environment in two experimental contexts. In a given context, the results were consistent with previous work that described the transformation as a second-order linear system. Classically, the parameters of such second-order models are associated with physical properties of the limb such as viscosity and stiffness that are commonly assumed to be approximately constant. By contrast, we found that the fit kernels differed strongly across tasks in these and other parameters, suggesting context-dependent changes in neural and biomechanical processes. We additionally fit residuals to a simple noise model and found that the form of the noise was highly conserved across both contexts and animals. Strikingly, the fitted noise also closely matched that found previously in a human steering task. Altogether, this work presents a kernel-based analysis that characterizes the context-dependence of deterministic and non-deterministic components of a closed-loop sensorimotor task.

Selective Effects of Ongoing Alpha-Band Activity on Magno- and Parvo-Mediated Detection
Spontaneous fluctuations in cortical excitability, as reflected in variation in occipital alpha-band activity (8-12 Hz), have been shown to explain trial-to-trial variability in perception. Specifically, observers typically report seeing a stimulus more often during states of weak alpha power, likely due to a shift in detection criterion. However, prior work has paid little attention to the specific stimulus properties mediating detection. In early vision, different stimulus properties are preferentially processed along the magnocellular (MC) and parvocellular (PC) pathways, which vary in their preference for spatial and temporal frequency content and chromatic information. The goal of this study was to understand how spontaneous alpha power affects the detection of stimuli which are preferentially processed by either the MC or PC pathway. To achieve this, we used the "Steady/Pulsed Paradigm" which presented a brief, near-threshold stimulus in two conditions intended to bias processing to one or the other pathway. Our results showed an interaction effect of pre-stimulus alpha power on detection between the two conditions. While weak alpha power was predictive of seeing the stimulus in the steady condition (MC-biased), no significant effect was found in the pulsed condition (PC-biased). This interaction was driven by a selective alpha-related criterion shift in the steady condition, with no effect of pre-stimulus alpha on sensitivity (d') in either condition. Our results imply that alpha oscillations may differentially regulate excitability in the MC and PC pathways.

Single field evolution rule governs the dynamics of representational drift in mouse hippocampal dorsal CA1 region.
How the brain reconciles dynamism with stability to balance learning and reliable memory storage has not yet been fully understood. To address the critical question, we longitudinally recorded place cells in the hippocampal dorsal CA1 region over 7 to 56 days, utilizing multiple goal-oriented navigation paradigms across various environments. We found that over 80% of place cells displayed multiple fields, undergoing complex evolution events including field disappearance, formation, and retention. Place fields from the same neuron showed limited coordination (~5%), with a preference for synchronized changes. We further uncovered the single field evolution rule: the longer a field remains active, the more likely it is to continue being active; conversely, the longer a field remains inactive, the less likely it is to recover the future fate of a place field depends on its past activity. Mathematical modeling revealed that this rule sufficiently demonstrates the growing stability of the dCA1 spatial representation at the population level.

Encoding of antennal position and velocity by the Johnstons organ in hawkmoths
Insect antennae function as versatile, multimodal sensory probes in diverse behavioural contexts. In addition to their primary role as olfactory organs, they serve essential mechanosensory functions across insects, including auditory perception, vestibular feedback, airflow detection, gravity sensing, and tactile sensation. These diverse functions are facilitated by the mechanosensory Johnstons organ (JO), located at the joint between the second antennal segment, known as the pedicel, and the flagellum. The pedicel-flagellum joint lacks muscles which means that the Johnstons organs can perceive only passive deflections of the flagellum. Earlier work which characterized the sensitivity and short response time of the sensory units of JO in hawkmoths, showed that their sensitivity to a broad frequency range is range-fractionated. This vastly expands the functional repertoire of the JO. However, it is not clear what components of antennal kinematics are encoded by the JO. Here, we conducted experiments to test the hypothesis that JO neurons encode the position and velocity of angular movements of the flagellum. We recorded intracellularly from the axons of primary sensory neurons of JO while stimulating it with ramp-and-hold stimuli in which antennal position or antennal angular velocity was maintained at various constant values. Our study shows that JO neurons encode angular velocity and position of the antenna in their response. We also characterized the neural adaptation of the responses to angular velocities and positions. A majority of neurons were sensitive to a movement in the ventrad direction, in the direction of gravity. The adaptation and the directional response properties give rise to a nonlinear hysteresis-like response. Together, these findings highlight the neurophysiological basis underlying the functional versatility of the JO.

Imaging of developing human brains with ex vivo PSOCT and dMRI
The human brain undergoes substantial developmental changes in the first five years of life. Particularly in the white matter, myelination of axons occurs near birth and continues at a rapid pace during the first 2 to 3 years. Diffusion MRI (dMRI) has revolutionized our understanding of developmental trajectories in white matter. However, the mm-resolution of in vivo techniques bears significant limitation in revealing the microstructure of the developing brain. Polarization sensitive optical coherence tomography (PSOCT) is a three-dimensional (3D) optical imaging technique that uses polarized light interferometry to target myelinated fiber tracts with micrometer resolution. Previous studies have shown that PSOCT contributes significantly to the elucidation of myelin content and quantification of fiber orientation in adult human brains. In this study, we utilized the PSOCT technique to study developing brains during the first 5 years of life in combination with ex vivo dMRI. The results showed that the optical properties of PSOCT quantitatively reveal the myelination process in young children. The imaging contrast of the optic axis orientation is a sensitive measure of fiber orientations in largely unmyelinated brains as young as 3-months-old. The micrometer resolution of PSOCT provides substantially enriched information about complex fiber networks and complements submillimeter dMRI. This new optical tool offers great potential to reveal the white matter structures in normal neurodevelopment and developmental disorders in unprecedented detail.

Dopamine reveals adaptive learning of actions representation
Efficient decision-making requires two key processes: learning values from actions and identifying a set of relevant actions to learn from in a given context. While dopamine (DA) is a well-known substrate for signaling reward prediction errors (RPEs) from selected actions to adjust behavior, the process of establishing and switching between action representations is still poorly understood. To address this gap, we used fiber photometry and computational modelling in a three-armed bandit task where mice learned to seek rewards delivered through three successive rule sets, displaying distinct strategies in each rule. We show that DA dynamically reflected RPEs computed from different task features, revealing context-specific internal representations. Our findings demonstrate that mice not only learned and updated action values but also action representations, adapting the features from which they learn across rules for flexible adjustment of their decision strategy.

Synaptogyrin-3 Prevents Cocaine Addiction and Dopamine Deficits
Synaptogyrin-3, a functionally obscure synaptic vesicle protein, interacts with vesicular monoamine and dopamine transporters, bringing together dopamine release and reuptake sites. Synaptogyrin-3 was reduced by chronic cocaine exposure in both humans and rats, and was inversely correlated with motivation to take cocaine in rats. Synaptogyrin-3 overexpression in dopamine neurons reduced cocaine self-administration, decreased anxiety-like behavior, and enhanced cognitive flexibility. Overexpression also enhanced nucleus accumbens dopamine signaling and prevented cocaine-induced deficits, suggesting a putative therapeutic role for synaptogyrin-3 in cocaine use disorder.

Abnormal multisensory facilitation patterns relate to disorganized thinking severity and cognitive decline in schizophrenia
Past research has demonstrated that patients with schizophrenia (SP) have visual processing and multisensory integration deficits. Additional studies report that sensory abnormalities are related to positive symptoms. To further understand how multisensory abnormalities relate to positive symptoms, we administered a multisensory integration task requiring the evaluation of perceived distance from auditory, visual, and multisensory stimuli with varying synchrony as well as clinical and neurocognitive assessments. Overall, patients had greater facilitation than healthy controls and the near synchronous condition had the most facilitation in comparison to other conditions. To further examine how multisensory facilitation relates to symptom severity, we performed a Ward cluster analysis that grouped participants by their multisensory facilitation profile. In contrast to what was expected, none of the Ward clusters were populated by a single group. Patients in cluster 3 had a significantly greater disorganization factor score than those in cluster 1. Our in-depth comparison between Ward clusters and neuropsychological tests reveal patients with greater multisensory facilitation experience the most cognitive deficits. Overall, our results demonstrate that multisensory integration is related to behavioral and cognitive deficits in complex ways. Further research is needed to understand the relationship between multisensory integration and schizophrenia symptomology.

Evaluating phasic transcutaneous vagus nerve stimulation (taVNS) with pupil dilation: the importance of stimulation intensity and sensory perception
The efficacy of transcutaneous auricular vagus nerve stimulation (taVNS) as a non-invasive method to modulate physiological markers of noradrenergic activity of the Locus Coeruleus (LC), such as pupil dilation, is increasingly more discussed. However, taVNS studies show high heterogeneity of stimulation effects. Therefore, a taVNS setup was established here to test different frequencies (10 Hz and 25 Hz) and intensities (3 mA and 5 mA) during phasic stimulation (3 s) with time-synchronous recording of pupil dilation in younger adults. Specifically, phasic real taVNS and higher intensity led to increased pupil dilation, which is consistent with phasic invasive VNS studies in animals. The results also suggest that the influence of intensity on pupil dilation may be stronger than that of frequency. However, there was an attenuation of taVNS-induced pupil dilation when differences in perception of sensations were considered. Specifically, pupil dilation during phasic stimulation increased with perceived stimulation intensity. The extent to which the effect of taVNS induces pupil dilation and the involvement of sensory perception in the stimulation process are discussed here and require more extensive research. Additionally, it is crucial to strive for comparable stimulation sensations during systematic parameter testing in order to investigate possible effects of phasic taVNS on pupil dilation in more detail.

Proliferative arrest induces neuron differentiation and innate immune responses in control and Creutzfeldt-Jakob Disease agent infected rat septal neurons
Rat post-mitotic septal (SEP) neurons, engineered to conditionally proliferate at 330C, differentiate when arrested at 37.50C and can be maintained for weeks without cytotoxic effects. Nine independent cDNA libraries were made to follow arrest-induced neural differentiation and innate immune responses in normal (Nl) uninfected and CJ agent infected SEP cells. Proliferating Nl versus latently infected (CJ-) cells showed few RNA-seq differences. However arrest induced major changes. Normal cells displayed a plethora of anti-proliferative transcripts. Additionally, known neuron differentiation transcripts, e.g., Agtr2, Neuregulin-1, GDF6, SFRP4 and Prnp were upregulated. These Nl neurons also displayed many activated IFN innate immune genes, e.g., OAS1, RTP4, ISG20, GTB4, CD80 and cytokines, complement, and clusterin (CLU) that binds to misfolded proteins. In contrast, arrested highly infectious CJ+ cells (10 logs/gm) downregulated many replication controls. Furthermore, arrested CJ+ cells suppressed neuronal differentiation transcripts, including Prnp which is essential for CJ agent infection. CJ+ cells also enhanced IFN stimulated pathways, and analysis of the 342 CJ+ unique transcripts revealed additional innate immune and anti-viral-linked transcripts (e.g., Il17, ISG15, and RSAD2 (viperin)). This data show: 1) innate immune transcripts are produced by normal neurons during differentiation; 2) CJ infection can enhance and expand anti-viral responses; 3) latent CJ infection epigenetically imprints many proliferative pathways to thwart complete arrest. CJ+ brain microglia, white blood cells and intestinal myeloid cells with shared transcripts may be stimulated to educe latent CJD infections that can be clinically silent for >30 years.

Fundamental limitations of kilohertz-frequency carriers in afferent fiber recruitment with transcutaneous spinal cord stimulation
The use of kilohertz-frequency (KHF) waveforms has rapidly gained momentum in transcutaneous spinal cord stimulation (tSCS) to restore motor function after paralysis. However, the mechanisms by which these fast-alternating currents depolarize efferent and afferent fibers remain unknown. Our study fills this research gap by providing a hypothesis- and evidence-based investigation using peripheral nerve stimulation, lumbar tSCS, and cervical tSCS in 25 unimpaired participants together with computational modeling. Peripheral nerve stimulation experiments and computational modeling showed that KHF waveforms negatively impact the processes required to elicit action potentials, thereby increasing response thresholds and biasing the recruitment towards efferent fibers. While these results translate to tSCS, we also demonstrate that lumbar tSCS results in the preferential recruitment of afferent fibers, while cervical tSCS favors recruitment of efferent fibers. Given the assumed importance of proprioceptive afferents in motor recovery, our work suggests that the use of KHF waveforms should be reconsidered to maximize neurorehabilitation outcomes, particularly for cervical tSCS. We posit that careful analysis of the mechanisms that mediate responses elicited by novel approaches in tSCS is crucial to understanding their potential to restore motor function after paralysis.

Early life psychosocial stress increases binge-like ethanol consumption and alters microglia and brain macrophage population density in early adulthood
As key regulators of brain homeostasis, microglia and brain macrophages are vital to development. However, early life stress (ELS) experiences, like bullying, can have lasting consequences on microglial function. During ELS, microglia and brain macrophages alter their engagement with cells and synapses. These alterations can result in lasting adaptations in circuit function that could lead to an increased risk to develop maladaptive drinking behaviors and alcohol use disorder (AUD) in adulthood. Whether microglia and brain macrophages truly mediate the relationship between ELS and AUD remains elusive. Here, we report: 1) a novel early life psychosocial stress exposure, PSS, increases binge-like ethanol consumption in early adulthood. 2) This increase in ethanol drinking is associated with alterations in microglia and brain macrophage populations in the ventral hippocampus and across other brain regions. 3) Inhibiting microglia and brain macrophages using a CSF1-r inhibitor, GW2580, results in a trend towards a reduction in binge-like ethanol consumption and normalizes microglia and brain macrophage populations. Therefore, our study suggests that acutely inhibiting microglia and brain macrophage function during periods of early life psychosocial stress may help reduce the risk to develop risky drinking behaviors and AUD.

Maintenance of taste receptor cell presynaptic sites requires gustatory nerve fibers
The turnover and re-establishment of peripheral taste synapses is vital to maintain connectivity between the primary taste receptor cells and the gustatory neurons which relay taste information from the tongue to the brain. Despite the importance of neuron-taste cell reconnection, mechanisms governing synapse assembly and the specificity of synaptic connections is largely unknown. Here we use the expression of presynaptic proteins, CALHM1 and Bassoon, to probe whether nerve fiber connectivity is an initiating factor for the recruitment of presynaptic machinery in different populations of taste cells. Under homeostatic conditions, the vast majority (>90%) of presynaptic sites are directly adjacent to nerve fibers. In the days immediately following gustatory nerve transection and complete denervation, Bassoon and CALHM1 puncta are markedly reduced. This suggests that nerve fiber innervation is crucial for the recruitment and maintenance of presynaptic sites. In support of this, we find that expression of Bassoon and Calhm1 mRNA transcripts are significantly reduced after denervation. During nerve fiber regeneration into the taste bud, presynaptic sites begin to replenish, but are not as frequently connected to nerve fibers as intact controls (~50% compared to >90%). This suggests that gustatory neuron proximity, rather than direct contact, likely drives taste receptor cells to express and aggregate presynaptic proteins at the cell membrane. Together, these data support the idea that trophic factors secreted by gustatory nerve fibers prompt taste receptor cells to produce presynaptic specializations at the cell membrane, which in turn may guide neurons to form mature synapses. These findings provide new insights into the mechanisms driving synaptogenesis and synaptic plasticity within the rapidly changing taste bud environment.

Differential Patterns of Gut and Oral Microbiomes in Hispanic Individuals with Cognitive Impairment
Alzheimer's disease and related dementias (ADRD) have been associated with alterations in both oral and gut microbiomes. While extensive research has focused on the role of gut dysbiosis in ADRD, the contribution of the oral microbiome remains relatively understudied. Furthermore, the potential synergistic interactions between oral and gut microbiomes in ADRD pathology are largely unexplored. This study aims to evaluate distinct patterns and potential synergistic effects of oral and gut microbiomes in a cohort of predominantly Hispanic individuals with cognitive impairment (CI) and without cognitive impairment (NC). We conducted 16S rRNA gene sequencing on stool and saliva samples from 32 participants (17 CI, 15 NC; 62.5% female, mean age = 70.4 {+/-} 6.2 years) recruited in San Antonio, Texas, USA. Correlation analysis through MaAslin2 assessed the relationship between participants' clinical measurements (e.g., fasting glucose and blood cholesterol) and their gut and saliva microbial contents. Differential abundance analysis evaluated taxa with significant differences between CI and NC groups, and alpha and beta diversity metrics assessed within-sample and group compositional differences. Our analyses revealed no significant differences between NC and CI groups in fasting glucose or blood cholesterol levels. However, a clear association was observed between gut microbiome composition and levels of fasting glucose and blood cholesterol. While alpha and beta diversity metrics showed no significant differences between CI and NC groups, differential abundance analysis revealed an increased presence of oral genera such as Dialister, Fretibacterium, and Mycoplasma in CI participants. Conversely, CI individuals exhibited a decreased abundance of gut genera, including Shuttleworthia, Holdemania, and Subdoligranulum, which are known for their anti-inflammatory properties. No evidence was found for synergistic contributions between oral and gut microbiomes in the context of ADRD. Our findings suggest that similar to the gut microbiome, the oral microbiome undergoes significant modifications as individuals transition from NC to CI. Notably, the identified oral microbes have been previously associated with periodontal diseases and gingivitis. These results underscore the necessity for further investigations with larger sample sizes to validate our findings and elucidate the complex interplay between oral and gut microbiomes in ADRD pathogenesis.

Characterizing microglial signaling dynamics during inflammation using single-cell mass cytometry
Microglia play a critical role in maintaining central nervous system (CNS) homeostasis and display remarkable plasticity in their response to inflammatory stimuli. However, the specific signaling profiles that microglia adopt during such challenges remain incompletely understood. Traditional transcriptomic approaches provide valuable insights, but fail to capture dynamic post-translational changes. In this study, we utilized time-resolved single-cell mass cytometry (CyTOF) to measure distinct signaling pathways activated in microglia upon exposure to bacterial and viral mimetics - lipopolysaccharide (LPS) and polyinosinic-polycytidylic acid (Poly(I:C)), respectively. Furthermore, we evaluated the immunomodulatory role of astrocytes on microglial signaling in mixed cultures. Microglia or mixed cultures derived from neonatal mice were treated with LPS or Poly(I:C) for 48 hrs. Cultures were stained with a panel of 33 metal-conjugated antibodies targeting signaling and identity markers. High-dimensional clustering analysis was used to identify emergent signaling modules. We found that LPS treatment led to more robust early activation of pp38, pERK, pRSK, and pCREB compared to Poly(I:C). Despite these differences, both LPS and Poly(I:C) upregulated the classical activation markers CD40 and CD86 at later time-points. Strikingly, the presence of astrocytes significantly blunted microglial responses to both stimuli, particularly dampening CD40 upregulation. Our studies demonstrate that single-cell mass cytometry effectively captures the dynamic signaling landscape of microglia under pro-inflammatory conditions. This approach may pave the way for targeted therapeutic investigations of various neuroinflammatory disorders. Moreover, our findings underscore the necessity of considering cellular context, such as astrocyte presence, in interpreting microglial behavior during inflammation.

From circuits to lifespan: translating mouse and human timelines with neuroimaging based tractography
Age is a major predictor of developmental processes and disease risk, but humans and model systems (e.g., mice) differ substantially in the pace of development and aging. The timeline of human developmental circuits is well known. It is unclear how such timelines compare to those in mice. We lack age alignments across the lifespan of mice and humans. Here, we build upon our Translating Time resource, which is a tool that equates corresponding ages during development. We collected 477 time points (n=1,132 observations) from age-related changes in body, bone, dental, and brain processes to equate corresponding ages across humans and mice. We acquired high- resolution diffusion MR scans of mouse brains (n=12) at sequential stages of postnatal development (postnatal day 3, 4, 12, 21, 60) to trace the timeline of brain circuit maturation (e.g., olfactory association pathway, corpus callosum). We found heterogeneity in white matter pathway growth. The corpus callosum largely ceases to grow days after birth while the olfactory association pathway grows through P60. We found that a P3 mouse equates to a human at roughly GW24, and a P60 mouse equates to a human in teenage years. Therefore, white matter pathway maturation is extended in mice as it is in humans, but there are species-specific adaptations. For example, olfactory-related wiring is protracted in mice, which is linked to their reliance on olfaction. Our findings underscore the importance of translational tools to map common and species-specific biological processes from model systems to humans.

Cognitive sequences in obsessive-compulsive disorder are supported by frontal cortex ramping activity and mediated by symptom severity
Completing sequences is a part of everyday life. Many such sequences can be considered abstract - that is, defined by a rule that governs the order but not the identity of individual steps (e.g., getting dressed for work). Over-engagement in ritualistic and repetitive behaviors seen in obsessive-compulsive disorder (OCD) suggests that abstract sequences may be disrupted in this disorder. Previous work has shown the necessity of the rostrolateral prefrontal cortex (RLPFC) for abstract sequence processing and that neural activity increases (ramps) in this region across sequences (Desrochers, Chatham, & Badre, 2015; Desrochers, Collins, & Badre, 2019). Neurobiological models of the cortico-striatal-thalamo-cortical (CSTC) loops describe prefrontal circuitry connected to RLPFC and that is believed to be dysfunctional in OCD. As a potential extension of these models, we hypothesized that neural dynamics of RLPFC could be disrupted in OCD during abstract sequence engagement. We found that neural dynamics in RLPFC did not differ between OCD and healthy controls (HCs), but that increased ramping in pregenual anterior cingulate cortex (pACC), and superior frontal sulcus (SFS) dissociates these two groups in an abstract sequence paradigm. Further, we found that anxiety and depression symptoms mediated the relationship between observed neural activity and behavioral differences observed in the task. This study highlights the importance of investigating ramping as a relevant neural dynamic during sequences and suggests expansion of current neurobiological models to include regions that support sequential behavior in OCD. Further, our results may point to novel regions to consider for neuromodulatory treatments of OCD in the future.

Monkey lateral prefrontal cortex subregions differentiate between perceptual exposure to visual stimuli
In everyday life, humans must parse visual stimuli with highly variable amounts of perceptual experience, ranging from incredibly familiar to entirely new. Even when choosing a novel to buy at a bookstore, one is exposed to covers they have seen numerous times intermixed with recently released titles. Visual exposure to stimuli is known to have distinct neural correlates in the lateral prefrontal cortex (LPFC) of nonhuman primates. However, it is currently unknown if this function may be localized to specific subregions within the LPFC. Specifically, we aimed to determine whether the posterior fundus of area 46 (p46f), an area that responds to deviations from a learned sequence, also responds to less frequently presented stimuli outside of the sequential context. We compare responses in p46f to the adjacent subregion, posterior ventral area 46 (p46v), which we propose may be more likely to show exposure-dependent responses due to its proximity to known novelty responsive regions. To test whether p46f or p46v represent perceptual exposure, we performed awake functional magnetic resonance imaging (fMRI) on three male monkeys as they observed visual stimuli that varied in their number of daily presentations. Here we show that p46v, but not p46f, shows preferential activation to stimuli with low perceptual exposure, which further localizes exposure-dependent effects in monkey LPFC. These results align with previous research that has found novelty responses in ventral LPFC and are consistent with the designation of p46f as having a sequence-specific function. Further, they expand on our knowledge of the specific role of LPFC subregions and localize perceptual exposure processing within this broader brain region.

Lifespan Oscillatory Dynamics in Lexical Production: A Population-based MEG Resting-State Analysis
Lexical production remains relatively preserved across the lifespan, but cognitive control demands increase with age to support efficient semantic access. It suggests a domain-general and a language-specific component. Current neurocognitive models suggest the Default Mode Network (DMN) may drive the interplay between these components, impacting the trajectory of production performance with a pivotal shift around midlife. However, the corresponding time-varying architecture still needs clarification. Here, we leveraged MEG resting-state data from healthy adults aged 18-88 from a CamCAN population-based sample. We found that DMN temporal dynamics shift from anterior-ventral to posterior-dorsal states until midlife to mitigate word-finding challenges. Similarly, sensorimotor integration along this posterior path enhances cross-talk with lower-level circuitry as the dynamic information flow with more anterior, higher-order cognitive states gets compromised. It suggests a bottom-up, exploitation-based form of cognitive control in the aging brain, highlighting the interplay between abstraction, control, and perceptive-motor systems in preserving lexical production.

Deliberative Behaviors and Prefrontal-Hippocampal Coupling are Disrupted in a Rat Model of Fetal Alcohol Spectrum Disorders
Fetal alcohol spectrum disorders (FASDs) are characterized by a range of physical, cognitive, and behavioral impairments. Determining how temporally specific alcohol exposure (AE) affects neural circuits is crucial to understanding the FASD phenotype. Third trimester AE can be modeled in rats by administering alcohol during the first two postnatal weeks, which damages the medial prefrontal cortex (mPFC), thalamic nucleus reuniens, and hippocampus (HPC), structures whose functional interactions are required for working memory and executive function. Therefore, we hypothesized that AE during this period would impair working memory, disrupt choice behaviors, and alter mPFC-HPC oscillatory synchrony. To test this hypothesis, we recorded local field potentials from the mPFC and dorsal HPC as AE and sham intubated (SI) rats performed a spatial working memory task in adulthood and implemented algorithms to detect vicarious trial and errors (VTEs), behaviors associated with deliberative decision-making. We found that, compared to the SI group, the AE group performed fewer VTEs and demonstrated a disturbed relationship between VTEs and choice outcomes, while spatial working memory was unimpaired. This behavioral disruption was accompanied by alterations to mPFC and HPC oscillatory activity in the theta and beta bands, respectively, and a reduced prevalence of mPFC-HPC synchronous events. When trained on multiple behavioral variables, a machine learning algorithm could accurately predict whether rats were in the AE or SI group, thus characterizing a potential phenotype following third trimester AE. Together, these findings indicate that third trimester AE disrupts mPFC-HPC oscillatory interactions and choice behaviors.

Using DeepLabCut-Live to probe state dependent neural circuits of behavior with closed-loop optogenetic stimulation
Background: Closed-loop behavior paradigms enable us to dissect the state-dependent neural circuits underlying behavior in real-time. However, studying context-dependent locomotor perturbations has been challenging due to limitations in molecular tools and techniques for real-time manipulation of spinal cord circuits. New Method: We developed a novel closed-loop optogenetic stimulation paradigm that utilizes DeepLabCut-Live pose estimation to manipulate primary sensory afferent activity at specific phases of the locomotor cycle in mice. A compact DeepLabCut model was trained to track hindlimb kinematics in real-time and integrated into the Bonsai visual programming framework. This allowed an LED to be triggered to photo-stimulate sensory neurons expressing channelrhodopsin at user-defined pose-based criteria, such as during the stance or swing phase. Results: Optogenetic activation of nociceptive TRPV1+ sensory neurons during treadmill locomotion reliably evoked paw withdrawal responses. Photoactivation during stance generated a brief withdrawal, while stimulation during swing elicited a prolonged response likely engaging stumbling corrective reflexes. Comparison with Existing Methods: This new method allows for high spatiotemporal precision in manipulating spinal circuits based on the phase of the locomotor cycle. Unlike previous approaches, this closed-loop system can control for the state-dependent nature of sensorimotor responses during locomotion. Conclusions: Integrating DeepLabCut-Live with optogenetics provides a powerful new approach to dissect the context-dependent role of sensory feedback and spinal interneurons in modulating locomotion. This technique opens new avenues for uncovering the neural substrates of state-dependent behaviors and has broad applicability for studies of real-time closed-loop manipulation based on pose estimation.

Nomination of a novel plasma protein biomarker panel capable of classifying Alzheimer's disease dementia with high accuracy in an African American cohort
Introduction: African Americans (AA) are widely understudied in plasma biomarker studies for Alzheimer's disease (AD) and current diagnostic biomarker candidates do not reflect the heterogeneity of AD. Methods: Untargeted proteome measurements were obtained using the SomaScan 7k platform to identify novel plasma biomarkers for AD in a cohort of AA clinically diagnosed as AD dementia (n=183) or cognitively unimpaired (CU, n=145). Machine learning approaches were implemented to identify the set of plasma proteins that achieve the best classification accuracy. Results: A plasma protein panel achieved an area under the curve (AUC) of 0.91 to classify AD dementia vs CU. The reproducibility of this finding was observed in the ANMerge plasma and AMP-AD Diversity brain datasets (AUC=0.83; AUC=0.94). Discussion: This study demonstrates the potential of biomarker discovery through untargeted plasma proteomics and machine learning approaches. Our findings also highlight the potential importance of the matrisome and cerebrovascular dysfunction in AD pathophysiology.