Biology subjects

Brown, R. E.

Publications and source records attributed to Brown, R. E..

18 recordsLinked to original sources

bioRxiv2026

Sustained Activation of a Defensive Behavior-Related Circuit Elicits a distinct Homeostatic Sleep Response

Animals and humans often forgo sleep in response to other physiological needs. Prolonged wakefulness leads to a compensatory homeostatic sleep response involving enhanced amounts of sleep and electroencephalographic delta (0.5-4 Hz) power, yet how the type of preceding wakefulness and brain activity affects this response is poorly understood. Here we compared the pattern of brain activity and the homeostatic sleep response following sustained wakefulness induced by either gentle handling (GH), a widely used form of sleep deprivation in rodents; or optogenetic stimulation of neurons involved in avoidance/defensive responses, basal forebrain (BF) vesicular glutamatergic transporter type 2 (vGluT2) neurons. Local infusion of ionotropic glutamate receptor antagonists partially attenuated the wake-inducing effect of vGluT2-stimulation, but not that of GH, suggesting mediation by other non-glutamatergic BF subtypes. These antagonists also tended to produce more NREM sleep and a shorter NREM latency in the recovery period following vGluT2-stimulation. Both manipulations increased high theta (7-9 Hz) during wakefulness, an effect partially blocked by local infusion of the glutamate antagonist cocktail. Enhanced NREM sleep time and NREM delta power in the recovery period were more pronounced following GH than following vGluT2 stimulation. Whole-brain cFos immunohistochemistry revealed stronger neuronal activity in brain regions involved in stress and avoidance/defensive behaviors with vGluT2 stimulation, compared to GH. Our results demonstrate that activation of a brain system involved in defensive behavior (BF vGluT2) elicits a distinct and diminished homeostatic sleep response compared to GH, potentially an evolutionary sleep adaptation which would maintain vigilance in the face of an ongoing threat.

neuroscience↗

bioRxiv2026

Early Loss of Deep Restorative Sleep and Auditory Stimulus Evoked 40-Hz activity of Hippocampal Parvalbumin Neurons in the APP/PS1 Mouse Model of Alzheimer's Disease

Sleep abnormalities and dysfunction of gamma band (30-80 Hz) activity generated by parvalbumin (PV) interneurons are early characteristics of Alzheimers disease (AD) which correlate with the severity of amyloid-{beta} deposition (A{beta}) and cognitive impairment. However, the timing of these alterations in vivo with respect to disease progression is unclear. Here, in longitudinal recordings from APP/PS1/PV-cre (AD mice) from 3-6 months, we found reduced sleep slow-wave power (0.5-4 Hz) in hippocampus and medial prefrontal cortex in AD mice as young as 3 months old, compared to non-AD (PV-cre) mice, well before overt pathology. This finding was primarily due to reductions in the NREM delta range (1.5-4 Hz), a hallmark of restorative functions of sleep. In contrast, beta (15-30 Hz) power linked to insomnia was significantly higher across all sleep-wake states. Loss of deep NREM sleep was not compensated by an increase in NREM sleep time, instead NREM sleep during the dark (active) phase was slightly but significantly lower in AD mice. 40-Hz auditory steady-state responses and associated evoked calcium responses of hippocampal PV neurons recorded using fiber photometry were also impaired by 3 months old. However, Y-maze performance in 3- and 6-month-old AD mice was not significantly different from non-AD mice. These results reveal reduced deep sleep and PV-associated 40-Hz activity as very early changes amenable to early intervention occurring prior to cognitive deficits. Furthermore, they establish APP/PS1 mice as a good model to causally test the relationship between sleep, PV neuronal activity and amyloid-mediated pathology.

neuroscience↗

bioRxiv2026

Synaptic GABA dysfunction of thalamocortical neurons impairs sleep spindle morphology and recovery from fearful memories.

The neural mechanisms which control the characteristic waxing and waning shape of sleep spindles and their dysregulation in neuropsychiatric illnesses are unresolved. Recent, sparse research shows post-traumatic stress disorder (PTSD) presents with abnormal spindle morphology and dysfunctional synaptic GABAA receptors in midline thalamic regions. We modeled this GABAA receptor dysfunction by localized CRISPR-Cas9-knockdown in mice. In contrast to a control group with intact GABAA receptors, mice with synaptic GABAA receptor knock-down in thalamocortical (TC) neurons lost the characteristic waxing and waning shape of sleep spindles and exhibited a failure to diminish conditioned contextual fear responses. Our results suggest that abnormally shaped sleep spindles may be a marker of synaptic GABA dysfunction in TC neurons and an indicator of disrupted fear extinction.

neuroscience↗

bioRxiv2026

Ecology of mosquitoes in Scottish wetlands and confirmation of Culex molestus and hybrids in Scotland

BackgroundThe recent spread of mosquito-borne pathogens and their vectors within Europe highlights the impact of climate change on vector-borne disease (VBD) distributions. Mosquito surveillance has been implemented in many European countries to monitor expansion of vector populations and VBDs, but ability to predict disease risk is constrained by geographic data gaps, particularly in northern areas. In the United Kingdom, wetland mosquito surveillance has been conducted extensively in England, with a knowledge deficit for Scotland. Here, we addressed this gap through a nationwide survey of mosquitoes at Scottish wetlands, with aims of 1) confirming the geographic distribution and environmental drivers of mosquito occurrence and abundance, and 2) identifying the presence of vector species of epidemiological concern, including the Culex pipiens molestus biotype, an important vector of emerging VBDs in mainland Europe. MethodsMonthly mosquito sampling was conducted between June and October 2023 at 22 sites across Scotland comprising six wetland types: coastal saltmarsh, wet grassland, wet woodland, reedbeds, ponds and blanket bog. Adult and larval populations were sampled using Biogents BG-Pro traps and larval dipping respectively. Microclimatic and hydrological variables were recorded at collection sites and used in generalised linear mixed models to identify predictors of mosquito presence and abundance. Results1951 adults (17 species/ groups) and 860 larvae (six species/ groups) were collected from wetlands over 183 and 164 sampling events respectively. Mosquitoes were widely distributed across the Scottish mainland including up to the northern coast, being found at all but one site. Several potential vector species including Culex pipiens s.l. and Anopheles claviger were common. Amongst the adult Culex pipiens s.l. specimens, approximately 8% were Culex pipiens biotype molestus or hybrid forms. Total mosquito abundance and that of key vector species were positively associated with temperature and rainfall. ConclusionsWe report the widespread distribution of mosquitoes in wetlands throughout Scotland, including potential vector species previously unconfirmed in Scotland. Predicted associations between mosquito abundance, rainfall and temperature indicate that climate change could favour mosquito populations in Scotland. Our results provide the first comprehensive description of mosquito ecology in Scotland, as required to update assessment of VBD risk under climate change.

ecology↗

bioRxiv2025

Interval Timing is altered in male Nrxn1+/- mice: A Model of Autism Spectrum Disorder

Autism spectrum disorder (ASD) is characterised by impaired social interactions and communication and increased repetitive and stereotypical behaviour. Neuroimaging shows functional abnormalities in brain areas involved in temporal processing of autistic individuals, and autistic individuals show deficits in interval timing. Neurexin (NRXN) mutations have been identified in a wide variety of neuropsychiatric disorders, including ASD, and Nrxn1+/- mice possess a mutation that disrupts the , {beta}, and {gamma} isoforms of Nrxn1, a gene involved in synapse structure. We investigated the interval timing abilities of the Nrxn1+/- mouse model of ASD in the peak interval procedure using a 15-second target interval and compared their performance with that of Nrxn1+/+ and Nrxn1{Delta}S5/- rescue mice. Two-month-old male Nrxn1+/+ (C57BL/6J), Nrxn1+/-, and Nrxn1{Delta}S5/-, mice were trained to obtain sucrose liquid rewards 15s after the onset of a discriminative stimulus (discrete fixed-interval training), and their timing responses were tested in non-reinforced probe trials. Our analysis of responses in individual trials revealed that Nrxn1+/- mice had overall earlier timing responses. This difference was manifested as earlier termination of responding in terms of the response curves. These findings are consistent with leftward shifts observed with experimental animal models of ASD. In conclusion, we believe that these results are indicative of a biased long-term memory in the Nrxn1+/- mouse model of ASD and may capture the timing deficit observed in autistic individuals. Lay SummaryNeurexins help nerve cells connect and communicate with each other, and changes in these genes are often seen in people with autism. Mice with a change in their neurexin 1 gene, called Nrxn1+/- mice, show autism-like behaviours. In a test that involves judging time, these mice respond early, similar to some people with autism. This study helps develop our understanding of how interval timing is affected in ASD.

neuroscience↗

bioRxiv2025

Motor deficits in the McGill-R-Thy1-APP transgenic rat model of Alzheimers Disease

The McGill-R-Thy1-APP rat is a transgenic model of Alzheimers Disease (AD) which expresses APP with two mutations found in cases of familial AD, resulting in the development of amyloid pathology and cognitive deficits. Motor deficits are common symptoms of AD, emerging early in the disease, and are correlated with AD neuropathology and cognitive symptoms. This study evaluated heterozygous and homozygous McGill-R-Thy1-APP rats and their wildtype littermates for spontaneous alternation and locomotion in the T and Y mazes, and motor behaviour on an accelerating rotarod. Because rats often jumped off the rotarod, the maximum latency to fall across trials was examined. We found no genotype or sex effects in spontaneous alternation in either maze, nor a significant correlation of spontaneous alternation behaviour between the mazes. Female rats travelled greater distances than male rats in both mazes. While there was no genotype effect in the T maze on distance travelled, in the Y maze the hemizygous rats travelled shorter distances than the wildtype rats, while the homozygous rats travelled greater distances. There was a significant correlation between the distances travelled in each maze. Both heterozygous and homozygous rats performed worse than their wildtype littermates on the rotarod, while heavier rats performed worse than lighter rats, and female rats performed worse than male rats once their differences in weights were accounted for. These findings support the continued use of these rats as a model of AD and highlight the need to consider the possible confounding effect motor impairments have on other behavioural tests.

animal behavior and cognition↗

bioRxiv2024

Individualized temporal patterns dominate cortical upstate and sleep depth in driving human sleep spindle timing

Sleep spindles are critical for memory consolidation and strongly linked to neurological disease and aging. Despite their significance, the relative influences of factors like sleep depth, cortical up/down states, and spindle temporal patterns on individual spindle production remain poorly understood. Moreover, spindle temporal patterns are typically ignored in favor of an average spindle rate. Here, we analyze spindle dynamics in 1008 participants from the Multi-Ethnic Study of Atherosclerosis using a point process framework. Results reveal fingerprint-like temporal patterns, characterized by a refractory period followed by a period of increased spindle activity, which are highly individualized yet consistent night-to-night. We observe increased timing variability with age and distinct gender/age differences. Strikingly, and in contrast to the prevailing notion, individualized spindle patterns are the dominant determinant of spindle timing, accounting for over 70% of the statistical deviance explained by all of the factors we assessed, surpassing the contribution of slow oscillation (SO) phase ([~]14%) and sleep depth ([~]16%). Furthermore, we show spindle/SO coupling dynamics with sleep depth are preserved across age, with a global negative shift towards the SO rising slope. These findings offer novel mechanistic insights into spindle dynamics with direct experimental implications and applications to individualized electroencephalography biomarker identification.

neuroscience↗

bioRxiv2023

Sleep-Deep-Net learns sleep wake scoring from the end-user and completes each record in their style.

Sleep-wake scoring is a time-consuming, tedious but essential component of clinical and pre-clinical sleep research. Sleep scoring is even more laborious and challenging in rodents due to the smaller EEG amplitude differences between states and the rapid state transitions which necessitate scoring in shorter epochs. Although many automated rodent sleep scoring methods exist, they do not perform as well when scoring new data sets, especially those which involve changes in the EEG/EMG profile. Thus, manual scoring by expert scorers remains the gold-standard. Here we take a different approach to this problem by using a neural network to accelerate the scoring of expert scorers. Sleep-Deep-Net (SDN) creates a bespoke deep convolution neural network model for individual electroencephalographic or local-field-potential records via transfer learning of GoogleNet, by learning from a small subset of manual scores of each EEG/LFP record as provided by the end-user. SDN then automates scoring of the remainder of the EEG/LFP record. A novel REM scoring correction procedure further enhanced accuracy. SDN reliably scores EEG and LFP data and retains sleep-wake architecture in wild-type mice, in sleep induced by the hypnotic zolpidem, in a mouse model of Alzheimers disease and in a genetic knock-down study, when compared to manual scoring. SDN reduced manual scoring time to 1/12. Since SDN uses transfer learning on each independent recording, it is not biased by previously scored existing data sets. Thus, we find SDN performs well when used on signals altered by a drug, disease model or genetic modification. STATEMENT OF SIGNIFICANCESleep medicine is often critically advanced by translational research based on in vivo electrophysiologic mouse data. A necessary but time-consuming step in this field is scoring epochs of recordings into wakefulness, non-rapid-eye-movement sleep and non-rapid-eye-movement sleep. Despite efforts to automate this, manual scoring remains the gold-standard since automatic methods poorly handle data that is not similar enough to data used during development. Here, we describe a novel automated sleep scoring method that involves retraining a deep-convolution-neural-net capable of computer vision to score sleep-wake patterns after learning from a small set of manual scores within a record. This avoids biasing the model to expect data to be the same as its training set from previous records.

neuroscience↗

bioRxiv2023

Neuronal PAS domain 1 identifies a major subpopulation of wakefulness-promoting GABAergic neurons in basal forebrain

Here we describe a novel group of basal forebrain (BF) neurons expressing neuronal PAS domain 1 (Npas1), a developmental transcription factor linked to neuropsychiatric disorders. Immunohistochemical staining in Npas1-cre-2A-TdTomato mice revealed BF Npas1+ neurons are distinct from well-studied parvalbumin or cholinergic neurons. Npas1 staining in GAD67-GFP knock-in mice confirmed that the vast majority of Npas1+ neurons are GABAergic, with minimal colocalization with glutamatergic neurons in vGlut1-cre-tdTomato or vGlut2-cre-tdTomato mice. The density of Npas1+ neurons was high, 5-6 times that of neighboring cholinergic, parvalbumin or glutamatergic neurons. Anterograde tracing identified prominent projections of BF Npas1+ neurons to brain regions involved in sleep-wake control, motivated behaviors and olfaction such as the lateral hypothalamus, lateral habenula, nucleus accumbens shell, ventral tegmental area and olfactory bulb. Chemogenetic activation of BF Npas1+ neurons in the light (inactive) period increased the amount of wakefulness and the latency to sleep for 2-3 hr, due to an increase in long wake bouts and short NREM sleep bouts. Non-REM slow-wave (0-1.5 Hz) and sigma (9-15 Hz) power, as well as sleep spindle density, amplitude and duration, were reduced, reminiscent of findings in several neuropsychiatric disorders. Together with previous findings implicating BF Npas1+ neurons in stress responsiveness, the anatomical projections of BF Npas1+ neurons and the effect of activating them suggest a possible role for BF Npas1+ neurons in motivationally-driven wakefulness and stress-induced insomnia. Identification of this major subpopulation of BF GABAergic neurons will facilitate studies of their role in sleep disorders, dementia and other neuropsychiatric conditions involving BF. SIGNIFICANCE STATEMENTWe characterize a group of basal forebrain (BF) neurons in the mouse expressing neuronal PAS domain 1 (Npas1), a developmental transcription factor linked to neuropsychiatric disorders. BF Npas1+ neurons are a major subset of GABAergic neurons distinct and more numerous than cholinergic, parvalbumin or glutamate neurons. BF Npas1+ neurons target brain areas involved in arousal, motivation and olfaction. Activation of BF Npas1+ neurons in the light (inactive) period increased wakefulness and the latency to sleep due to increased long wake bouts. Non-REM sleep slow waves and spindles were reduced reminiscent of findings in several neuropsychiatric disorders. Identification of this major subpopulation of BF GABAergic wake-promoting neurons will allow studies of their role in insomnia, dementia and other conditions involving BF.

neuroscience↗

bioRxiv2021

THO and TRAMP complexes prevent transcription-replication conflicts, DNA breaks, and CAG repeat contractions

Expansion of structure-forming CAG/CTG repetitive sequences is the cause of several neurodegenerative disorders and deletion of repeats is a potential therapeutic strategy. Transcription-associated mechanisms are known to cause CAG repeat instability. In this study, we discovered that Thp2, an RNA export factor and member of the THO complex, and Trf4, a key component of the TRAMP complex involved in nuclear RNA degradation, are necessary to prevent CAG fragility and repeat contractions in a S. cerevisiae model system. Depletion of both Thp2 and Trf4 proteins causes a highly synergistic increase in CAG repeat fragility, indicating a complementary role of the THO and TRAMP complexes in preventing genome instability. Loss of either Thp2 or Trf4 causes an increase in RNA polymerase stalling at the CAG repeats and genome-wide transcription-replication conflicts (TRCs), implicating impairment of transcription elongation as a cause of CAG fragility and instability in their absence. Analysis of the effect of RNase H1 overexpression on CAG fragility and TRCs suggests that co-transcriptional R-loops are the main cause of CAG fragility in the thp2{Delta} mutants. In contrast, CAG fragility and TRCs in the trf4{Delta} mutant can be compensated for by RPA overexpression, suggesting that excess unprocessed RNA in TRAMP4 mutants leads to reduced RPA availability and high levels of TRCs. Our results show the importance of RNA surveillance pathways in preventing RNAPII stalling, TRCs, and DNA breaks, and show that RNA export and RNA decay factors work collaboratively to maintain genome stability.

genetics↗

bioRxiv2021

MTG16 (CBFA2T3) represses E protein-dependent transcription to regulate colonic secretory cell differentiation, epithelial regeneration, and tumorigenesis

Aberrant epithelial differentiation and regeneration contribute to colon pathologies including inflammatory bowel disease (IBD) and colitis-associated cancer (CAC). MTG16 (CBFA2T3) is a transcriptional corepressor expressed in the colonic epithelium. MTG16 deficiency in mice exacerbates colitis and increases tumor burden in CAC, though the underlying mechanisms remain unclear. Here, we identified MTG16 as a central mediator of epithelial differentiation, promoting goblet and restraining enteroendocrine cell development in homeostasis and enabling regeneration following dextran sulfate sodium (DSS)-induced colitis. Transcriptomic analyses implicated increased E box-binding transcription factor (E protein) activity in MTG16-deficient colon crypts. Using a novel mouse model with a point mutation that disrupts MTG16:E protein complex formation (Mtg16P209T), we established that MTG16 exerts control over colonic epithelial differentiation and regeneration by repressing E protein-mediated transcription. Mimicking murine colitis, MTG16 expression was increased in biopsies from patients with active IBD compared to unaffected controls. Finally, uncoupling MTG16:E protein interactions only partially phenocopied the enhanced tumorigenicity of Mtg16-/- colon in the azoxymethane(AOM)/DSS-induced model of CAC, indicating that MTG16 protects from tumorigenesis through additional mechanisms. Collectively, our results demonstrate that MTG16, via its repression of E protein targets, is a key regulator of cell fate decisions during colon homeostasis, colitis, and cancer. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC="FIGDIR/small/467178v3_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@21948borg.highwire.dtl.DTLVardef@9f60eeorg.highwire.dtl.DTLVardef@194b9fdorg.highwire.dtl.DTLVardef@13eb1a7_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2021

Basal forebrain parvalbumin neurons modulate vigilant attention.

Attention is impaired in many neuropsychiatric disorders1 and by sleep disruption, leading to decreased workplace productivity and increased risk of accidents2-4. Thus, understanding the underlying neural substrates is important for developing treatments. The basal forebrain (BF) is a brain region which degenerates in dementia5-7 and is implicated in the negative effects of sleep disruption on vigilance and cognition8,9. Previous studies demonstrated that the BF controls cortical fast oscillations that underlie attention10-12 and revealed the important role of cholinergic neurons13-15. However, the role of other neurochemically defined BF subtypes is unknown. Recent work has shown that one population of BF GABAergic neurons containing the calcium-binding protein parvalbumin (PV) control cortical fast oscillations and arousals from sleep16-19 but their role in awake behavior is unclear. Thus, here we test the hypothesis that BF-PV neurons modulate vigilant attention in mice. A lever release version of the rodent psychomotor vigilance test (rPVT) was used to assess vigilant attention as measured by reaction time. Brief and continuous low power optogenetic excitation of BF-PV neurons (1s,473nm@5mW) that preceded the cue light signal by 0.5s improved vigilant attention as indicated by quicker reaction times. In contrast, both sleep deprivation (8h) and optogenetic inhibition of BF-PV neurons (1s,530nm@10mW) slowed reaction times. Importantly, BF-PV excitation rescued the reaction time deficits in sleep deprived mice. These findings reveal for the first time a role for BF-PV neurons in attention. HIGHLIGHTSO_LIOptogenetic methods tested the neural circuitry of vigilant attention in mice C_LIO_LIExcitation of basal forebrain parvalbumin neurons quickened reaction times C_LIO_LISleep deprivation or inhibition of parvalbumin neurons slowed reaction times C_LIO_LIExcitation of parvalbumin neurons rescued deficits produced by sleep deprivation C_LI

neuroscience↗

bioRxiv2020

CRISPR knockdown of GABAA alpha3 subunits on thalamic reticular neurons enhances deep sleep

Identification of mechanisms which increase deep sleep could lead to novel treatments which promote the restorative effects of sleep. Here, knockdown of the 3 GABAA-receptor subunit from parvalbumin neurons in the thalamic reticular nucleus using CRISPR-Cas9 gene editing increased the thalamocortical delta oscillations implicated in many health-promoting effects of sleep. Inhibitory synaptic currents were strongly reduced in vitro. Effects were most pronounced in the mouse sleep (light) period. Further analysis identified a novel deep-sleep state in mice prior to NREM-REM transitions which was preferentially affected by deletion of 3 subunits. Our results identify a functional role for GABAA receptors on Thalamic Reticular Nucleus neurons and suggest antagonism of 3 subunits as a strategy to enhance deep sleep. One Sentence SummarySelective genetic knockdown of the major subunit of GABAA receptors present in the thalamic reticular nucleus enhanced deep sleep in mice.

neuroscience↗

bioRxiv2020

Cell Programmed Nutrient Partitioning in the Tumor Microenvironment

The tumor microenvironment (TME) includes transformed cancer and infiltrating immune cells1,2. Cancer cells can consume large quantities of glucose through Warburg metabolism3,4 that can be visualized with positron emission tomography (PET). While infiltrating immune cells also rely on glucose, disruptions to metabolism can contribute to tumor immunological evasion5-9. How immune cell metabolism is programmed or restrained by competition with cancer cells for nutrients, remains uncertain. Here we used PET tracers to measure the accessibility of glucose and glutamine to cell subsets in the TME. Surprisingly, myeloid cells including macrophages were the greatest consumers of intra-tumoral glucose, followed by T cells and cancer cells. Cancer cells, in contrast, had the highest glutamine uptake. This distinct nutrient partitioning was programmed through selective mTORC1 signaling and glucose or glutamine-related gene expression. Inhibition of glutamine uptake enhanced glucose uptake across tumor resident cell types and shifted macrophage phenotype, demonstrating glucose is not limiting in the TME. Thus, cancer cells are not the only cells in tumors which exhibit high glucose uptake in vivo and instead preferentially utilize glutamine over other cell types. We observe that intrinsic cellular programs can play a major role in the use of some nutrients. Together, these data argue cell selective partitioning of glucose and glutamine can be exploited to develop therapies and imaging strategies to alter the metabolic programs of specific cell populations in the TME.

cancer biology↗

bioRxiv2020

The Phase Shift Correlation function uncovers periodic Ulcerative Colitis microbiome relationships without temporal sampling.

Correlation analysis is a fundamental technique to determine potential relationships within biological processes. However, many biological processes have been shown to function in a periodic manner. When modeling correlations, the fluctuations that are associated with periodicity cause significant issues. We have implemented a Phase Shift Correlation (PSC) algorithm, with a corresponding value PSCrho, to address the periodicity and phase variance associated with features that vary with the same frequency -- but are phase shifted. The phase shift could well indicate causality with one features quantitative change leading to the change in the other feature. We applied the PSC algorithm to Ulcerative Colitis (UC) microbiome data and compared the resulting feature relationships with the equivalent Spearman correlation function results. PSC located many instances of higher phase shifted correlations, where the corresponding Pearson correlation was low.

bioinformatics↗

bioRxiv2020

Characterization of basal forebrain glutamate neurons suggests a role in control of arousal and avoidance behavior

The basal forebrain (BF) is involved in arousal, attention, and reward processing but the role of individual BF subtypes is still being uncovered. Glutamatergic neurons are the least well-understood of the three major BF neurotransmitter phenotypes. Here we analyzed the distribution, size, calcium-binding protein content and projections of the major group of BF glutamate neurons expressing the vesicular glutamate transporter subtype 2 (vGluT2) and tested the functional effect of activating them. Mice expressing Cre recombinase under control of the vGluT2 promoter were crossed with a reporter strain expressing the red fluorescent protein, tdTomato, to generate vGluT2-cre-tdTomato mice. Immunohistochemical staining for choline acetyltransferase and a cross with mice expressing green fluorescent protein selectively in GABAergic neurons confirmed cholinergic, GABAergic and vGluT2+ neurons represent separate BF subpopulations. Subsets of BF vGluT2+ neurons expressed the calcium binding proteins calbindin or calretinin, suggesting that multiple subtypes of BF vGluT2+ neurons exist. Anterograde tracing using adeno-associated viral vectors expressing channelrhodopsin2- enhanced yellow fluorescent fusion proteins revealed major projections of BF vGluT2+ neurons to neighboring BF cholinergic and parvalbumin neurons, as well as to extra-BF areas involved in the control of arousal and regions responding to aversive or rewarding stimuli such as the lateral habenula and ventral tegmental area. Optogenetic activation of BF vGluT2 neurons in a place preference paradigm elicited a striking avoidance of the area where stimulation was given. Together with previous optogenetic findings suggesting an arousal-promoting role, our findings suggest BF vGluT2 neurons play a dual role in promoting wakefulness and avoidance behavior.

neuroscience↗

bioRxiv2019

Basal forebrain parvalbumin neurons mediate arousals from sleep induced by hypercarbia or auditory stimuli

Brief arousals from sleep in patients with sleep apnea and other disorders prevent restful sleep, and contribute to cognitive, metabolic and physiologic dysfunction. Little is currently known about which neural systems mediate these brief arousals, hindering the development of treatments. The basal forebrain (BF) receives inputs from many nuclei of the ascending arousal system. These inputs include the brainstem parabrachial neurons which promote arousal in response to elevated blood carbon dioxide levels, as seen in sleep apnea. Optical inhibition of the terminals of parabrachial neurons in the BF impairs cortical arousals to hypercarbia, but which cell types within the BF mediate cortical arousals in response to hypercarbia or other sensory stimuli is unknown. Here using optogenetic techniques in mice, we show that BF parvalbumin (PV) neurons fulfill several criteria for a system mediating brief arousals from sleep. Optical stimulation of BF PV neurons during the light period, when mice normally sleep, caused rapid transitions to wakefulness and increased wake bout durations. Unlike many other ascending arousal systems, arousals induced by stimulation of BF PV neurons were brief, resulting in only a small (13.6%) increase in the total amount of wakefulness. Bilateral optical inhibition of BF PV neurons increased the latency to arousal produced by hypercarbia or auditory stimuli. Thus, BF PV neurons are an important component of the brain circuitry which generates brief arousals from sleep in response to internal and external sensory stimuli.

neuroscience↗

bioRxiv2019

Pathfinder: open source software for analyzing spatial navigation search strategies

Spatial navigation is a universal behavior that varies depending on goals, experience and available sensory stimuli. Spatial navigational tasks are routinely used to study learning, memory and goal-directed behavior, in both animals and humans. One popular paradigm for testing spatial memory is the Morris water maze, where subjects learn the location of a hidden platform that offers escape from a pool of water. Researchers typically express learning as a function of the latency to escape, though this reveals little about the underlying navigational strategies. Recently, a number of studies have begun to classify water maze search strategies in order to clarify the precise spatial and mnemonic functions of different brain regions, and to identify which aspects of spatial memory are disrupted in disease models. However, despite their usefulness, strategy analyses have not been widely adopted due to the lack of software to automate analyses. To address this need we developed Pathfinder, an open source application for analyzing spatial navigation behaviors. In a representative dataset, we show that Pathfinder effectively characterizes the development of highly-specific spatial search strategies as male and female mice learn a standard spatial water maze. Pathfinder can read data files from commercially- and freely-available software packages, is optimized for classifying search strategies in water maze paradigms, but can also be used to analyze 2D navigation by other species, and in other tasks, as long as timestamped xy coordinates are available. Pathfinder is simple to use, can automatically determine pool and platform geometry, generates heat maps, analyzes navigation with respect to multiple goal locations, and can be updated to accommodate future developments in spatial behavioral analyses. Given these features, Pathfinder may be a useful tool for studying how navigational strategies are regulated by the environment, depend on specific neural circuits, and are altered by pathology.

animal behavior and cognition↗