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Ferguson, R.

Publications and source records attributed to Ferguson, R..

13 recordsLinked to original sources

Self-initiated restrictive feeding: a new mouse model of restrictive eating disorders

Restrictive eating disorders are serious psychiatric illnesses carrying the highest morality of any mental health condition, yet their neurobiology remains poorly understood. Progress is constrained by a shortage of animal models with convincing construct validity. In particular, anorexia nervosa (AN), a self-initiated restriction of caloric intake, is notoriously difficult to replicate in mice, which would otherwise offer an ideal pathway to understanding the neural mechanisms of this strongly gene-linked disorder, similar to advances in other neuropsychiatric conditions. Here, we introduce a model of self-initiated restrictive feeding (SIRF), using open-source behavioural devices. We show that SIRF specifically affects female mice, persists in the absence of the initial triggering conditions, and correlates with social rank - features that parallel AN. Meal pattern analysis shows that SIRF arises alongside decreased meal size and duration, with meal frequency unaltered in the long-term. Chemogenetic inhibition of the insular cortex, a brain region consistently implicated in AN and interoceptive processing, reduced SIRF. In sum, this proof-of-principle study suggests that SIRF provides a tractable model of self-initiated restriction as a platform for dissecting the neural circuits that initiate and sustain restrictive eating.

neuroscience↗

Genomes of two arid-zone marsupials uncover contrasting responses to climatic change

Small marsupials in the family Dasyuridae are a key component of Australias arid and semi-arid fauna, whose high species richness is proposed to reflect an opportunity-driven adaptive radiation. Despite growing interest in this group from both ecological and evolutionary perspectives, genomic data for most species is non-existent, or limited to a few marker loci. Here, we generated a chromosome-level reference genome and a de novo mitochondrial genome for the desert-dwelling Wongai ningaui (Ningaui ridei). The nuclear genome assembly is highly contiguous, with a scaffold N50 of 594.5 MB and high BUSCO gene recovery (93.8%). Additionally, we produced a draft assembly for the related, semi-arid slender-tailed dunnart (Sminthopsis murina). We then used these assemblies to explore the demographic histories of these species. We find evidence for contrasting patterns of population growth during the late Pleistocene and early Holocene, corresponding with differences in local climate, potentially consistent with differences in optimal habitat. The new genomic resources and demographic findings presented here provide a foundation for future studies on adaptive specialisation in this group of Australian marsupials. Significance StatementDasyurid marsupials are the primary carnivorous and insectivorous mammals in Australia. This family includes species such as the endangered Tasmanian devil (Sarcophilus harrisii) and quolls (Genus Dasyurus), as well as an emerging model species, the fat-tailed dunnart (Sminthopsis crassicaudata). Despite the species richness within dasyurids, most species remain under-studied. This is particularly true of arid and semi-arid zone species, who are often small in size, live in remote habitats and are cryptic by nature. By creating genome assemblies for two dasyurid species, this study provides resources to support phylogenetic, comparative and conservation research in arid zone marsupials. Importantly, the studys finding that arid and semi-arid species show distinct trajectories of demographic change in response to historical climate may have implications for the resilience of locally-adapted dasyurid species to ongoing climate change.

genomics↗

3,500 years of sheeppox virus evolution inferred from archaeological and codicological genomes

Sheeppox virus (SPPV) is a major livestock pathogen causing economic hardship through reduced production and death of vulnerable sheep, with written descriptions of sheeppox-like disease recorded since antiquity. We report 21 novel ancient SPPV genomes spanning the Eurasian steppe Bronze Age ([~]1,700 BCE) to the Early Modern period in Western Europe, including multiple genomes obtained from medieval parchment. We estimate that major capripoxvirus lineages diverged [~]11,500-3,700 years ago, overlapping known translocations and bio-cultural developments in sheep. Our dataset supports SPPV diverging first within the lineage leading to goatpox virus and lumpy skin disease virus, and that known gene inactivation events within SPPV and goatpox virus occur in our earliest SPPV genomes. These findings reveal that the food security of Eurasian communities have been threatened by sheeppox for over 3,700 years, and provide new insights to the genomic evolution and potential host adaptation of sheeppox virus. TeaserAncient viral genomes from manuscripts and teeth illuminate the evolutionary history of sheeppox virus.

genetics↗

Evidence of sheeppox in 17th-19th century France: a multi-disciplinary investigation of a sheep mass mortality assemblage

Epizootic outbreaks posed major threats to food security, economic stability, and animal welfare in past communities. While these events and their impacts are well documented in historical records, very few archaeological examples are known, and none for which the causal agent has been reliably identified. We therefore investigated a 17 th-19th century ovine mass-mortality assemblage from Louvres, France, using an interdisciplinary approach integrating paleogenetics, zooarchaeology, history, and veterinary science. The deposit contained nine complete, articulated sheep skeletons, mostly older individuals of both sexes, which were unbutchered but displayed flaying marks. Aside from minor age-related lesions and one ectopic bone nodule, the animals exhibited good skeletal health, with no obvious cause of death. Ancient DNA analysis was therefore conducted and proved highly successful, conclusively identifying two ovine pathogens: sheeppox virus (SPPV) and the parasite Taenia hydatigena. When considered alongside archaeological, veterinary and historical evidence, these findings indicate that the animals died during a sheeppox outbreak. This study provides the first detection of SPPV in archaeological bone and the first paleogenome of this virus. It also offers new insights into livestock health, mortality, and disease management in early modern France, demonstrating the value of integrated approaches for reconstructing past epizootics.

genetics↗

Microglial activation and alpha-synuclein oligomers drive the early inflammatory phase of Parkinson's disease

Parkinsons disease (PD) is characterised by insoluble -synuclein (Syn) aggregates in Lewy bodies (LBs) within the substantia nigra, with cortical pathology appearing as the disease progresses. Late-stage LB deposition, cellular stress, and neuronal loss obscure disease-driving events, we therefore performed multi-regional transcriptomic and aggregate profiling in early-midstage PD brains (Braak 3-4), where cortical regions are pathologically unaffected. We report neuroimmune activation as an early PD feature, characterised by the expansion of a high-SNCA-expressing microglial state. This robust immune signature occurs prior to LB formation, but is associated with oligomeric Syn within cortical microglia. In hiPSC-derived microglia, both endogenous Syn oligomerisation, and exogenous oligomer uptake, trigger transcriptional reprogramming, characterised by interferon-driven inflammation, antigen presentation, and mitochondrial suppression, closely mirroring the early PD brain. These findings describe mechanisms by which Syn oligomerisation potently initiates early neuroinflammation, highlighting a critical interplay between proteinopathy and immune activation at the earliest stages of disease.

neuroscience↗

Single-molecule validation and optimized protocols for the use of secondary nanobodies in multiplexed immunoassays

Recently developed secondary nanobodies or single-domain antibodies present a powerful tool for immunodetection. Unlike traditional antibodies, their monovalence enables pre-association with primary antibodies prior to sample staining, presenting a straightforward affinity-based antibody labeling solution. This not only simplifies and streamlines immunoassays, it also supports multiplexed techniques where conflicts in the species of the desired primary antibodies preclude standard indirect immunostaining. Despite these advantages, the use of secondary nanobodies remains sparse, due perhaps to a lack of evaluation on their suitability for assays requiring quantification and an assessment of optimal protocols for their use. Here, we present a set of experiments spanning single-molecule detection to cell imaging that can be used to validate secondary nanobody binding, specificity, and their propensity for mis-targeted binding in multiplex assays. Using these tools, we analyzed the binding properties of commercially available secondary nanobodies and outline optimized protocols for their robust use.

biochemistry↗

Arbuscular mycorrhizal fungal genotype and nuclear organization as driving factors in host plant nutrient acquisition and stable carbon capture

O_LIArbuscular mycorrhizal fungi (AMF) are obligate root symbionts of most plants that improve plant growth by transferring nutrients into plant roots through networks of soil hyphae. These hyphal networks represent a carbon sink in soil; thus, it has been suggested that these fungi can also boost atmospheric carbon storage, highlighting their potential role in managing greenhouse emissions. In this study, we aimed to determine whether certain AMF genotypes and nuclear organizations (homokaryons vs heterokaryons) are associated with higher rates of host plant yield and carbon storage. C_LIO_LIWe compared Sudan-grass (Sorghum x drummondii) AMF inoculation across eight strains of Rhizophagus irregularis: four homokaryotic and four heterokaryotic strains. Sudan-grass was grown in a growth chamber, which included 13C-CO2 pulse labeling to track plant carbon into AMF. C_LIO_LIAMF inoculation increased total and belowground biomass, as well as phosphorous, magnesium, and manganese uptake in the host. Heterokaryons led to greater belowground biomass, as well as less variable increases in shoot phosphorous. Mycorrhizal inputs to soil mineral-associated organic carbon - a highly persistent carbon pool with slow turnover - were overall greater in heterokaryons than in homokaryons but varied significantly among strains. C_LIO_LIThis indicates that the potential for carbon storage by mycorrhizal carbon inputs varies based on fungal genomic identity and nuclear organization. Overall, inoculation improved the yield of Sudan-grass and resulted in significant inter-strain variation in persistent carbon contributions to the soil. This work highlights the importance of considering genotype and nuclear identity in assessments of AMF as bio-stimulants and drivers of carbon storage. C_LI Societal Impact StatementIt is crucial to develop strategies for reducing our continued excessive global increases in fertilizer applications and to offset CO2 emissions. The pervasive underground hyphal networks of arbuscular mycorrhizal fungi (AMF) present an enticing bio-stimulant and carbon sink. We inoculated Sudan-grass plants with eight genotypically distinct strains of a model AMF species to determine if strain identity affects plant growth and carbon storage. We found that plant biomass, nutrient acquisition and stable soil carbon inputs varied among strains, emphasizing the importance of AMF strain identity in the selection of AMF inoculants for optimizing crop yield and carbon storage.

microbiology↗

Geranylgeranylated-SCFFBXO10 Regulates Selective Outer Mitochondrial Membrane Proteostasis and Function

E3-ubiquitin ligases (E3s) are main components of the ubiquitin-proteasome system (UPS), as they determine substrate specificity in response to internal and external cues to regulate protein homeostasis. However, the regulation of membrane protein ubiquitination by E3s within distinct cell membrane compartments or organelles is not well understood. We show that FBXO10, the interchangeable component of the SKP1/CUL1/F-box ubiquitin ligase complex (SCF-E3), undergoes lipid-modification with geranylgeranyl isoprenoid at Cysteine953 (C953), facilitating its dynamic trafficking to the outer mitochondrial membrane (OMM). FBXO10 polypeptide does not contain a canonical mitochondrial targeting sequence (MTS); instead, its geranylgeranylation at C953 and the interaction with two cytosolic factors, PDE6{delta} (a prenyl group-binding protein), and HSP90 (a mitochondrial chaperone) orchestrate specific OMM targeting of prenyl-FBXO10 across diverse membrane compartments. The geranylgeranylation-deficient FBXO10(C953S) mutant redistributes away from the OMM, leading to impaired mitochondrial ATP production, decreased mitochondrial membrane potential, and increased mitochondrial fragmentation. Phosphoglycerate mutase 5 (PGAM5) was identified as a potential substrate of FBXO10 at the OMM using comparative quantitative mass spectrometry analyses of enriched mitochondria (LFQ-MS/MS), leveraging the redistribution of FBXO10(C953S). FBXO10, but not FBXO10(C953S), promoted polyubiquitylation and degradation of PGAM5. Examination of the role of this pathway in a physiological context revealed that the loss of FBXO10 or expression of prenylation-deficient-FBXO10(C953S) inhibited PGAM5 degradation, disrupted mitochondrial homeostasis, and impaired myogenic differentiation of human iPSCs and murine myoblasts. Our studies identify a mechanism for selective E3-ligase mediated regulation of mitochondrial membrane proteostasis and metabolic health, potentially amenable to therapeutic intervention.

cell biology↗

Therapeutic validation of MMR-associated genetic modifiers in a human ex vivo model of Huntington's disease

The pathological huntingtin (HTT) trinucleotide repeat underlying Huntingtons disease (HD) continues to expand throughout life. Repeat length correlates both with earlier age at onset (AaO) and faster progression, making slowing its expansion an attractive therapeutic approach. Genome-wide association studies have identified candidate variants associated with altered AaO and progression, with many found in DNA mismatch repair (MMR) associated genes. We examine whether lowering expression of these genes affects the rate of somatic expansion in human ex vivo models using HD iPSCs and HD iPSC-derived striatal neurons. We have generated a stable CRISPR interference HD iPSC line in which we can specifically and efficiently lower gene expression from a donor carrying over 125 CAG repeats. Lowering expression of each member of the MMR complexes MutS (MSH2, MSH3 & MSH6), MutL (MLH1, PMS1, PMS2 & MLH3) and LIG1 resulted in characteristic MMR deficiencies. Reduced MSH2, MSH3 and MLH1 slowed repeat expansion to the largest degree, while lowering either PMS1, PMS2 and MLH3 slowed it to a lesser degree. These effects were recapitulated in iPSC derived striatal cultures where MutL factor expression was lowered. Here, reducing the expression of MMR factors by CRISPRi to levels typically reached by current therapeutics effectively slows the pathogenic expansion of the HTT CAG repeat tract. We highlight members of the MutL family as potential therapeutic targets to slow repeat expansion with the aim to delay onset and progression of HD, and potentially other repeat expansion disorders exhibiting somatic instability. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/570095v1_ufig1.gif" ALT="Figure 1"> View larger version (64K): org.highwire.dtl.DTLVardef@13da0a4org.highwire.dtl.DTLVardef@fd831corg.highwire.dtl.DTLVardef@1aac392org.highwire.dtl.DTLVardef@de298_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

Modelling human endurance: Power laws vs critical power

The power-duration relationship describes the time to exhaustion for exercise at different intensities. It is generally believed to be a "fundamental bioenergetic property of living systems" that this relationship is hyperbolic. Indeed, the hyperbolic (a.k.a. critical-power) model which formalises this belief is the dominant tool for describing and predicting high-intensity exercise performance, e.g. in cycling, running, rowing, or swimming. However, the hyperbolic model is now the focus of two heated debates in the literature because: (a) it unrealistically represents efforts that are short (< 2 minutes) or long (> 15 minutes); (b) it contradicts widely-used performance predictors such as the so-called functional threshold power (FTP) in cycling. We contribute to both debates by demonstrating that the power-duration relationship is more adequately represented by an alternative, power-law model. In particular, we show that the often observed good fit of the hyperbolic model between 2 and 15 minutes should not be taken as proof that the power-duration relationship is hyperbolic. Rather, in this range, a hyperbolic function just happens to approximate a power law fairly well. We also prove mathematical results which suggest that the power-law model is a safer tool for pace selection than the hyperbolic model and that the former better models fatigue than the latter. Finally, we use the power-law model to shed light on popular performance predictors in cycling, running and rowing such as FTP and Jack Daniels "VDOT" calculator.

physiology↗

Structural Dynamics of Real and Modeled Solanum Stamens: Implications for Pollen Ejection by Buzzing Bees

An estimated 10% of flowering plant species conceal their pollen within tube-like anthers that dehisce through small apical pores (poricidal anthers). Bees extract pollen from poricidal anthers through a complex motor routine called floral buzzing, whereby the bee applies vibratory forces to the flower stamen by rapidly contracting its flight muscles. The resulting deformation depend on the stamens natural frequencies and vibration mode shapes, yet for most poricidal species these properties have not been sufficiently characterized. We performed experimental modal analysis on Solanum elaeagnifolium stamens to quantify their natural frequencies and vibration modes. Based on morphometric and dynamic measurements, we developed a finite element model of the stamen to identify how variable material properties, geometry and bee weight could affect its dynamics. In general, stamen natural frequencies fell outside the reported floral buzzing range, and variations in stamen geometry and material properties were unlikely to bring natural frequencies within this range. However, inclusion of bee mass reduced natural frequencies to within the floral buzzing frequency range and gave rise to an axial-bending vibration mode. We hypothesize that floral buzzing bees exploit the large vibration amplification factor of this mode to increase anther deformation, which may facilitate pollen ejection.

biophysics↗

Targeting Wnt Signaling To Overcome PARP Inhibitor Resistance

Epithelial ovarian cancer (EOC) has one of the highest deaths to incidence ratios. High grade serous ovarian carcinoma (HGSOC) is the most common and deadliest EOC histotype because of the lack of secondary therapeutic options following debulking surgery and platinum/taxane-based chemotherapies. For recurrent chemosensitive HGSOC, poly(ADP)-ribose polymerase inhibitors (PARPi; olaparib, rucaparib, or niraparib) represent an emerging treatment strategy. While PARPi are most effective in homologous recombination DNA repair-deficient (HRD) HGSOCs, more recent studies have observed a significant clinical benefit in non-HRD HGSOCs. However, all HGSOC patients are likely to acquire resistance to PARPi. Therefore, there is an urgent clinical need to better understand PARPi resistance, and to introduce novel combinatorial therapies to overcome PARPi resistance and extend HGSOC disease-free intervals. Utilizing a two BRCA2-mutated and one BRCA-wildtype HGSOC cell lines that are olaparib sensitive, we established resistant cells. Transcriptome analysis of the matched olaparib-sensitive versus resistant cells did not detect BRCA2 reversion mutations, but revealed activation of Wnt/TCF signaling pathway, as TCF transcriptional activity was significantly increased in PARPi-resistant cells. In parallel, forced activation of Wnt signaling in PARPi-sensitive cells via WNT3A stimulation reduced response to PARPi. In a recurrent-HGSOC PARPi insensitive patient-derived xenograft model there was an increase in a Wnt/TCF transcriptional target. PARPi resistant cells were sensitive to inhibition of Wnt signaling using the FDA-approved compound, pyrvinium pamoate, which has been shown to inhibit Wnt signaling. We observed that combining pyrvinium pamoate with olaparib resulted in a significant decrease in tumor burden and number of tumor nodules. This study demonstrates that Wnt signaling can mediate PARPi resistance in HGSOC and provides a clinical rationale for combining PARPi and Wnt inhibitors.

cancer biology↗

Nus factors have a widespread regulatory function in bacteria

Nus factors are broadly conserved across bacterial species, and are often essential for viability. A complex of five Nus factors (NusB, NusE, NusA, NusG and SuhB) is considered to be a dedicated regulator of ribosomal RNA folding, and has been shown to prevent Rho-dependent transcription termination. We have established the first cellular function for the Nus factor complex beyond regulation of ribosomal assembly: repression of the Nus factor-encoding gene, suhB. This repression occurs by translation inhibition followed by Rho-dependent transcription termination. Thus, the Nus factor complex can prevent or promote Rho activity depending on the gene context. Extensive conservation of NusB/E binding sites upstream of nus factor genes suggests that Nus factor autoregulation occurs in many species. Putative NusB/E binding sites are also found upstream of many other genes in diverse species, and we demonstrate Nus factor regulation of one such gene in Citrobacter koseri. We conclude that Nus factors have an evolutionarily widespread regulatory function beyond ribosomal RNA, and that they are often autoregulatory.

microbiology↗