Search bioRxiv⌕ Search

Biology subjects

Howard, C.

Publications and source records attributed to Howard, C..

13 recordsLinked to original sources

Hybridisation and herbivory fuel Amazonian tree radiations

Tropical rainforests, and Amazonia in particular, contain more tree species than anywhere else, most of which arose through rapid evolutionary radiations 1-3. Rapid radiations are often catalysed by ecological opportunity 4-6, which in rainforest trees is presented by intense insect herbivore pressure, spurring the evolution of novel plant defence chemistry to escape it 7. However, we do not understand how long-lived trees can adapt quickly enough to keep pace with rapidly-evolving insect herbivores. Here we show that hybridisation in rainforest trees, which was considered rare, allows exchange of gene clusters used in chemical defence against herbivore attack, facilitating rapid adaptation and diversification. Using genome sequencing for 461 individuals from the genus Inga, a characteristic Amazonian tree radiation, we find that regional tree communities form syngameons - networks of closely related, co-occurring species connected by gene flow. Integrating these genomes with herbivore abundance data from the same communities across the tropical Americas, we show that herbivore compositional turnover coincides with local, recurrent interspecific transfer of defence gene clusters that are retained by balancing selection, consistent with fluctuating selective pressure imposed by shifting herbivore communities. Together, our results demonstrate that hybridisation allows long-lived tropical trees to rapidly evolve chemical defences, fuelling adaptation to the relentless insect herbivory that structures the worlds most species-rich forests.

evolutionary biology↗

The role of integrins in T cell-mediated resistance to Cryptosporidium parvum

Cryptosporidium is a protozoan that infects epithelial cells of the small intestine and is a cause of diarrhea and death in immunocompromised individuals and malnourished children. Immunity to this parasite is mediated by an intestinal T cell response, which is generated in gut-associated lymphoid tissues and dependent on type 1 conventional dendritic cells (cDC1s). The initial priming of T cells is accompanied by changes in integrin expression and subsequent trafficking to the site of infection. The role of specific integrins in trafficking to the ileum during cryptosporidiosis is largely unknown. The development of a transgenic Cryptosporidium strain that expresses MHCI and MHCII-restricted model antigens provides the ability to track T cell responses to this parasite. Our studies in this system revealed marked changes in the integrin profile of parasite-specific T cells as they are activated and traffic to the gut, and demonstrate that cDC1s contribute to the expression of the integrins 4, {beta}7, {beta}1, and L. Surprisingly, blockade of the canonical gut-homing integrin 4{beta}7 does not impact the ability of parasite-specific T cells to access the gut. However, blockade of integrin L decreases the parasite-specific T cell frequency at the site of infection and delays control of parasite burden. These datasets highlight an 4{beta}7-independent mechanism of T cell trafficking to the small intestine and indicate that L is an integrin required for T cell-mediated resistance to Cryptosporidium.

immunology↗

A precise atlas of the human subcortex

Clinical interventions and neuroimaging in the subcortex require anatomical definitions that exceed the resolution and anatomical detail of currently available deformable brain atlases. Here, we introduce a high-resolution human brain atlas comprising 95 manually segmented grey and white matter structures as well as 82 white matter tracts compiled from a multitude of resources including ex-vivo MRI, histology, fibre dissections, and neuroanatomy textbooks. The atlas is defined at an isotropic resolution of 100 m and can be precisely deformed to individual subject brain anatomy. By providing precise definitions of both grey and white matter structures within and around the basal ganglia, thalamus, subthalamus, midbrain and cerebellum, the atlas provides a foundational resource for stereotactic surgery and subcortical brain imaging research, as well as for development of next-generation neuromodulation strategies.

neuroscience↗

Methylome and transcriptome mapping reveal miniscule DNA methyltransferase regulons in Salmonella enterica serovar Typhimurium

DNA methylation is a regulator of bacterial gene expression and adaptation, influencing traits such as virulence and antimicrobial resistance. The dynamic nature of DNA methylation enables rapid responses to changing environments and is a source of heterogeneity in bacterial populations. However, condition-dependent DNA methylation and consequences for transcriptional output remain poorly understood. We applied Oxford Nanopore sequencing to profile DNA methylation during exponential growth and late stationary phase of Salmonella enterica serovar Typhimurium and integrated these data with transcriptomic analyses. We found that each DNA methyltransferase (MTases) exhibits a distinct activity pattern across growth stages, which could not be explained by transcriptional levels of the corresponding enzymes. As predicted, DNA methylation patterns determined by regulatory MTases were dynamic across growth conditions whereas methylation patterns of MTases belonging to R-M systems were comparatively stable. We identified growth stage-specific methylation patterns for all studied MTases and correlations between methylation states and gene expression patterns. Together, these findings chart DNA methylation networks in the epigenetic regulation of bacterial physiology. Author summaryDNA methylation in bacteria is best known for its role protecting DNA from endonucleases, such as restriction-modification, and coordinating chromosome replication and mutation repair, yet DNA methylation also regulates gene expression and cell physiology. Previous studies primarily examined bacterial DNA methylation at single time points or in limited genomic regions, providing only a partial view of its biological significance. In this study, we used Oxford Nanopore sequencing to compare DNA methylation patterns in Salmonella enterica during exponential growth and late stationary phase then integrated these data with corresponding gene expression profiles. We identified numerous methylation target motifs, all of which demonstrated constitutively methylated or unmethylated regions. This systems-level analysis clarifies the role of DNA methylation in bacterial adaptation across growth stages and demonstrates the utility of Oxford Nanopore sequencing for genome-wide methylation profiling.

microbiology↗

Diverse infections transcriptionally reprogram the intestinal epithelium and epithelial-immune cell interactions

The distal small intestine plays vital roles in host physiology by regulating nutrient and fluid homeostasis. Despite being impacted in Crohns disease and a major target for a range of infections, we know relatively little about the complexity of cellular responses and cell-cell communication in the ileum during infection. Single cell and spatial transcriptomics have emerged as powerful technologies to study tissue heterogeneity in the gut, but these tools have focused on the large intestine, in part due to the accessibility of this tissue for biopsies and its importance in cancer. Here we present GutPath, an atlas of over 500,000 single cells with RNA and protein expression profiles for 91 cell states in the ileum across diverse infectious archetypes. We show that GutPath accurately captures established immune responses to infection while revealing pathogen-specific responses in enterocytes. To highlight the discovery potential of this atlas, we identify a novel enterocyte cell state present during Yersinia pseudotuberculosis infection that is spatially linked to bacterial load and tissue pathology. GutPath establishes a much-needed resource for the immunology community that will accelerate the study of the transcriptional diversity of cellular landscapes in the small intestine.

immunology↗

On the path to reference genomes for all biodiversity: lessons learned and laboratory protocols created in the Sanger Tree of Life core laboratory over the first 2000 species

Since its inception in 2019, the Tree of Life programme at the Wellcome Sanger Institute has released high-quality, chromosomally-resolved reference genome assemblies for over 2000 species. Tree of Life has at its core multiple teams, each of which are responsible for key components of the genome engine. One of these teams is the Tree of Life core laboratory, which is responsible for processing tissues across a wide range of species into high quality, high molecular weight DNA and intact RNA, and preparing tissues for Hi-C. Here, we detail the different workflows we have developed to successfully process a wide variety of species, covering plants, fungi, chordates, protists, arthropods, meiofauna and other metazoa. We summarise our success rates and describe how to best apply and combine the suite of current protocols, which are all publicly available at protocols.io.

molecular biology↗

Local and distributed information coding in the ventral stream

Neuroscience is replete with evidence that cognitive representations are distributed across many cortical regions. Yet, the scale and content of such distributed processing is unclear. Do findings of widespread information coding suggest a large-scale "forest" of regions interacting to represent information or instead imply a multitude of small-scale trees, processing information as localized modules. To investigate this distinction, we used visual and semantic representational analysis of fMRI data from 60 participants viewing everyday objects in multiple task contexts, and we examined the relationships between regions in terms of information coding. We demonstrate that coding of visual content in the occipital lobe is overwhelmingly modular, such that different occipital structures show limited coordination and tend to encode information redundantly. By contrast, the coding of semantic content in the inferior temporal lobe involves a high degree of coordination between regions, which optimize their coding to collectively represent a large semantic space with minimal redundancy between regions. No other brain area - neither the parietal nor prefrontal cortices - shows the preference for large-scale coding seen in the inferior temporal lobe. Taken together, these results outline a framework of how the ventral stream transitions from small-scale to large-scale coding as information progresses from visual to semantic representations. Significance statementHow does the brain convert incoming signals into usable information? Many studies have investigated this question by attempting to clarify which brain regions encode what information (e.g., V4 encodes color information). We instead aimed to shed light on the degree of coordination among information coding regions. We find that the visual-to-semantic transition as information flows anteriorly in the occipitotemporal cortex is accompanied by a shift from modular to distributed coding. That is, occipital regions encode perceptual information relatively independently with redundancy, while inferior temporal lobe regions cooperate to most efficiently represent a large space of semantic information. By leveraging ideas from information theory, our work introduces coding scale as a new dimension for understanding the architecture of information coding.

neuroscience↗

An updated DNA barcoding tool for Aloe vera and related CITES-regulated species

DNA barcoding has revolutionised the identification of illegally traded material of endangered species as it overcomes the lack of resolution encountered with morphological identification. Nonetheless, in recently evolved and highly diverse clades, such as the relatives of Aloe vera, the lack of interspecific sequence variation in standardised markers compromises the barcoding efficacy. We present a new DNA barcoding tool using 189 nuclear markers, optimised for aloes (Asphodelaceae, Alooideae). We built a comprehensive sequence reference dataset from taxonomically verified sources for >300 species and validated its reliability for identification using phylogenomic inference. Seven anonymised samples from verified botanical collections and ten plants seized at London Heathrow Airport were correctly identified to species level, including a critically endangered species from Madagascar. Commercially purchased samples were confirmed to be the species as advertised. An accurate, reliable DNA barcoding method for aloe identification introduces new assurance to regulatory processes for endangered plants in trade.

genomics↗

The Impact of Parabacteroides distasonis Colonization on Hosts' Microbiome, Metabolome, Immune Responses, and Diabetes Onset

Type 1 Diabetes (T1D) is a chronic disease caused by autoimmune destruction of insulin-producing pancreatic {beta}-cells. The insulin B-chain 9-23 (insB:9-23) peptide is established as a critical epitope in triggering T1D. In our previous study, we showed that Parabacteroides distasonis, a human gut commensal, contains an insB:9-23 mimic in its hprt protein (residues, 4-18). This mimic (hprt4-18) activates insB:9-23 specific T-cells, and colonization of P. distasonis in female NOD mice enhanced diabetes onset. Additionally, the presence of hprt:4-18 sequence in the gut microbiome is associated with seropositivity in infants. However, the impact of the colonization on the gut microbiome and intestinal immune cell compositions, gut permeability, cytokine, and serum metabolome profiles were unknown. Here, we addressed this gap using specific pathogen-free (SPF) and germ-free (GF) NOD mouse models. P. distasonis colonization had a minimal impact on gut microbiome composition and merely altered 28 ASVs upon colonization. In intraepithelial lymphocytes (IELs) of P. distasonis colonized SPF NOD mice, we observed a 1.72-fold reduction in T-helper cells and a 2.3-fold reduction in T-effector cells, along with a 1.85-fold reduction in B-cell populations. Further, P. distasonis did not alter serum metabolome and cytokine levels except for a decrease in IL-15. We observed no difference in the gene expression related to gut permeability. Similar to SPF mice, P. distasonis colonization in GF NOD mice induced severe insulitis without affecting gut permeability. On the other hand, P. distasonis lysate could induce insB:9-23 specific T cells. Altogether, these findings demonstrate that P. distasonis does not stimulate a nonspecific inflammatory immune response in the intestines, nor does it cause significant alterations in the gut microbiome, gut permeability, serum metabolome, or cytokine response. However, it does induce insulitis in GF NOD mice and activates insB:9-23 specific T-cells. These findings support our original hypothesis that P. distasonis colonization stimulates a specific immune response and enhances T1D onset in NOD mice via molecular mimicry.

immunology↗

Lesions that Cause Psychosis Map to a Common Brain Circuit in the Hippocampus

ImportanceIdentifying anatomy causally involved in psychosis could inform therapeutic neuromodulation targets for schizophrenia. ObjectiveTo assess whether lesions that cause secondary psychosis have functional connections to a common brain circuit. DesignThis case-control study mapped functional connections of published cases of lesions causing secondary psychosis compared with control lesions unassociated with psychosis. SettingThis study was conducted in a computational laboratory. ParticipantsPublished cases of lesion-induced psychosis were analyzed. Included subjects had documented brain lesions associated with new-onset psychotic symptoms without prior history of psychosis. Control cases included 1156 patients with lesions not associated with psychosis. Generalizability across lesional datasets was assessed using an independent cohort of 181 patients with brain lesions who subsequently underwent neurobehavioral testing. ExposuresLesions causing secondary psychosis. Main Outcomes and MeasuresPsychosis or no psychosis. Results153 lesions from published cases were determined to be causal of psychosis (65 [42%] male; mean [SD] age, 50.0 [20.8] years), 42 of which were described as "schizophrenia" or "schizophrenia-like". Lesions that caused secondary psychosis mapped to a common brain circuit defined by functional connectivity to the posterior subiculum of the hippocampus (84% functional overlap, pFWE<5 x 10-5). At a lower statistical threshold (75%> overlap, pFWE<5 x 10-4), this circuit included the ventral tegmental area, retrosplenial cortex, lobule IX and dentate nucleus of the cerebellum, and the mediodorsal and midline nuclei of the thalamus. This circuit was consistent when derived from "schizophrenia-like" cases (spatial r=0.98). We repeated these analyses after excluding lesions intersecting the hippocampus (n=47) and found a consistent functional connectivity profile (spatial r=0.98) with the posterior subiculum remaining the center of connectivity (>75% overlap, pFWE<5x10-5), demonstrating a circuit-level effect. In an independent observational cohort of patients with penetrating head trauma (n=181), lesions associated with symptoms of psychosis exhibited significantly similar connectivity profiles to the lesion-derived psychosis circuit (suspiciousness, p=0.025; unusual thought content, p=0.046). Voxels in the rostromedial prefrontal cortex (rmPFC) are highly correlated with this psychosis circuit (spatial r=0.82), suggesting the rmPFC as a promising TMS target for psychosis. Conclusions and RelevanceLesions that cause secondary psychosis affect a common brain circuit in the hippocampus. These results can help inform therapeutic neuromodulation targeting. Key PointsO_ST_ABSQuestionC_ST_ABSDo lesions that cause psychosis affect a common brain circuit? FindingsThis case-control study found that lesions causing psychosis specifically affected a common functional circuit aligning with the posterior subiculum of the hippocampus. This functional circuit was consistent across different psychotic symptoms, suggesting a shared neural substrate for psychosis. A similar circuit was derived when excluding lesions directly intersecting the hippocampus, indicating a circuit-level effect. MeaningIdentifying a common brain circuit causally involved in psychotic symptoms suggests that the hippocampus may be pivotal in the pathophysiology and treatment targeting of psychotic disorders.

neuroscience↗

Efferent Retargeting in Above-Knee Amputees is Positively Related to Phantom Limb Pain

Many individuals who undergo limb amputation experience persistent phantom limb pain (PLP). The underlying mechanism of PLP is unknown, but the phenomenon has been associated with reorganization in sensorimotor cortex following amputation. The traditional view is that cortical reorganization degrades the missing limbs representation. However, recent work suggests that an amputated limbs cortical representation remains intact and that reorganization reflects a retargeting of efferent projections to residual muscles proximal to the amputation site. Evidence of retargeting has only been shown in individuals with upper limb amputations, and the relationship of retargeting to PLP is controversial. This study assessed retargeting and its relationship to PLP in 10 individuals with lower limb amputations. We recorded electromyographic (EMG) activity in a residual thigh muscle (vastus lateralis, VL) in patients with above-knee amputations during cyclical movements of the foot. VL activity on the amputated side was compared to that recorded on patients intact side while they moved their phantom and intact feet, respectively. VL activity in the patient group was also compared to VL activity from a sample of 9 control participants with no amputation. We show that phantom foot movement is associated with greater VL activity in the amputated leg than that seen in the intact leg as well as that exhibited by controls. The magnitude of residual VL activity was also positively related to ratings of PLP. These results provide the first support for retargeting in lower limb amputees and suggest that retargeting is related to the experience of phantom pain. New and NoteworthyPrevious work has only examined retargeting in upper limb amputees. This study provides evidence for retargeting in lower limb amputees and suggests that retargeting is related to phantom limb pain.

neuroscience↗

It’s the fiber, not the fat: Significant effects of dietary challenge on the gut microbiome

BackgroundDietary effects on the gut microbiome has been shown to play a key role in the pathophysiology of behavioral dysregulation, inflammatory disorders, metabolic syndrome, and obesity. Often overlooked is that experimental diets vary significantly in the proportion and source of dietary fiber. Commonly, treatment comparisons are made between animals that are fed refined diets that lack soluble fiber and animals fed vivarium-provided chow diet that contain a rich source of soluble fiber. Despite the well-established role of soluble fiber on metabolism, immunity, and behavior via the gut microbiome, the extent to which measured outcomes may be driven by differences in dietary fiber is unclear. Further, the significant impact of sex and age in response to dietary challenge is likely important and should also be considered.\n\nResultsWe compared the impact of transitioning young and aged male and female mice from a chow diet to a refined low soluble fiber diet on body weight and gut microbiota. Then, to determine the contribution of dietary fat, we examined the impact of transitioning a subset of animals from refined low fat to refined high fat diet. Serial tracking of body weights revealed that consumption of low fat or high fat refined diet increased body weight in young and aged adult male mice. Young adult females showed resistance to body weight gain, while high fat diet-fed aged females had significant body weight gain. Transition from a chow diet to low soluble fiber refined diet accounted for most of the variance in community structure and composition across all groups. This dietary transition was characterized by a loss of taxa within the phylum Bacteroidetes and a concurrent bloom of Clostridia and Proteobacteria in a sex- and age-specific manner. Most notably, no changes to gut microbiota community structure and composition were observed between mice consuming either low- or high-fat diet, suggesting that transition to the refined diet that lacks soluble fiber is the primary driver of gut microbiota alterations, with limited additional impact of dietary fat on gut microbiota.\n\nConclusionCollectively, our results show that the choice of control diet has a significant impact on outcomes and interpretation related to body weight and gut microbiota. These data also have broad implications for rodent studies that draw comparisons between refined high fat diets and chow diets to examine dietary fat effects on metabolic, immune, behavioral, and neurobiological outcomes.

systems biology↗

Probability-based detection of phosphoproteomic uncertainty reveals rare signaling events driven by oncogenic kinase gene fusion

We describe a novel Bayesian method for estimating protein concentration and phosphorylation site occupancy ratios from mass spectrometry experiments. Our variance model assigns standard deviations to all quantitative ratios, even when only a single peptide is observed, increasing the number of quantifiable observations in a sample compared to conventional methods. We further demonstrate the application of this method using a dataset investigating the impact of the PRKAR1A-RET gene fusion in immortalized thyroid cells.

systems biology↗