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Gray, R.

Publications and source records attributed to Gray, R..

17 recordsLinked to original sources

Migratory bird and marine mammal surveillance fails to find evidence for an HPAI H5N1 2.3.4.4b incursion into Australia in 2025

The panzootic caused by high pathogenicity avian influenza (HPAI) H5N1 clade 2.3.4.4b has been devastating for animals, globally. Despite global spread, the virus remains absent in Oceania. Herein we report the results of our fourth year of enhanced migratory bird surveillance, coinciding with the spring migration of wild birds in 2025; none of the 847 migratory wild birds or 38 marine mammals were positive for HPAI H5N1, although we did detect LPAI. Surveillance remains a critical tool for HPAI H5N1 response, with early detection and rapid response being critical to mitigate the impacts of this virus on animal, environment and human health.

microbiology↗

Morphometric analyses of shape: The analysis software toolbox for quantification of craniofacial shape

Fetal alcohol spectrum disorders (FASD) are characterized by a varying set of physical, cognitive, and behavioral disabilities caused by prenatal ethanol exposure, including those to the facial skeleton. Ethanol-sensitive genetic loci contribute to this high degree of variation in FASD, which complicates analyses of facial shape. We have previously shown that we can analyze these changes in facial shape from gene-ethanol interactions in zebrafish. Zebrafish are an ideal model to analyze this variation for several reasons; 1. 70% of genes are orthologs between humans and zebrafish; 2. external fertilization allows researchers to control timing and dosage of ethanol treatments; 3. the structure of their facial skeletal is conserved with vertebrates; 4. translucent larvae enable direct viewing of changes to the craniofacial skeletal structure during development. However, analyzing the shape of the craniofacial skeleton can be difficult to fully assess through simple linear measures, as these do not capture overall changes in shape. In addition, changes in head size can complicate data interpretation. To address this, we undertook a morphometric approach, analyzing overall facial shape through principal component analyses via freeware software, TPSDigs2, MorphoJ, and PAST. The combination of this software allows for pairwise comparison of overall facial morphology. Here, we outline our approach and analysis of facial shape in ethanol-treated zebrafish mutants using these programs to conduct a series of complementary multivariate statistical analyses. SUMMARYThis work describes a protocol for quantifying craniofacial cartilage shape using free software (TPSDigs2, MorphoJ, and PAST) to measure changes in facial structure in zebrafish larvae.

developmental biology↗

Sequence based prediction of cell type specific microRNA binding and mRNA degradation for therapeutic discovery

MicroRNAs and RNA binding proteins are crucial elements of post-transcriptional gene regulation, which governs the fate of mRNA molecules in the cell. However, the landscape of these regulatory interactions, particularly across different mammalian cell types, remains underexplored. We describe REPRESS, a deep learning model that predicts cell-type-specific microRNA binding and mRNA degradation directly from RNA sequence. REPRESS was trained on AGO2-CLIP, miR-eCLIP and Degradome-Seq data profiling millions of microRNA binding and mRNA degradation sites across multiple cell types in human and mouse. It reveals biology that other state-of-the-art methods did not, such as identifying repressive non-canonical miRNA target sites and decoding the regulatory effects of sequence context and miRNA binding site multiplicity. REPRESS outperforms other advanced methods and neural architectures on a comprehensive suite of seven orthogonal tasks, including identifying genetic variants that affect microRNA binding, predicting out-of-distribution data from massively parallel reporter assays, and predicting canonical and non-canonical miRNA mediated repression. To demonstrate the general utility of REPRESS, we show that it provides insights into novel biology and the design of RNA therapeutics. Code is available at : https://github.com/deepgenomics/repress

genomics↗

Interactions between the Bone Morphogenetic Protein and the Planar Cell Polarity Pathways lead to distinctive ethanol-induced facial defects

BackgroundFetal Alcohol Spectrum Disorders (FASD) describes a spectrum of ethanol-induced neural and facial developmental defects. Ethanol susceptibility is modulated by genetics, but their underlying mechanisms remain poorly understood. In all vertebrates, a series of complex cellular events give rise to the body plan, including convergence & extension (C&E) and endoderm/ cranial neural crest (CNC-which gives rise to the facial skeleton) morphogenesis. These events are critical to establish complex signaling interactions, driving embryo development, including the facial skeleton. In zebrafish, C&E occurs between 6-10 hpf while endoderm/CNC morphogenesis occurs 10-24 hpf. Previous work shows that the PCP mutants are sensitive to ethanol from 6-24 hpf, covering both C&E and endoderm/CNC morphogenesis and exhibiting multiple defects to the forming head raising the question whether ethanol during both time windows drives PCP-ethanol defects. We hypothesize that PCP single and double mutants are ethanol sensitive 10-24 hpf, after C&E. We also hypothesize BMP signaling (sensitive 10-18 hpf) interacts with and sensitizes the PCP pathway to ethanol. MethodsHere, we treated PCP/BMP mutants with ethanol from 6-10, 10-18, 10-24 or 24-30 hpf and combined morphometric and linear measurements to examine facial development. ResultsWe show that PCP mutant larvae are ethanol-sensitive from 10-24 hpf, but not 6-10 or 24-30 hpf. We also show that BMP mutants sensitize PCP mutants to ethanol and lead to novel ethanol-independent midline craniofacial defects. Our results suggest that the ethanol-sensitive role of PCP pathway occurs after C&E, during endoderm/CNC morphogenesis and that the PCP and BMP pathways genetically interact during the morphogenesis events. ConclusionsUltimately, our work builds on a mechanistic paradigm of ethanol-induced birth defects we have been developing, connecting conceptual framework with concrete cellular events that could be ethanol-sensitive beyond facial development.

developmental biology↗

A Cry For Kelp: Evidence for Polyphenolic Inhibition of Oxford Nanopore Sequencing of Brown Algae

Genomic resources for macroalgae are increasingly important for conservation and commercial management, however the generation of such resources continues to be hampered by difficulties in the isolation of suitable DNA. Even when DNA has been isolated that otherwise appears high quality, such samples may not perform well during the sequencing process. We here compare Oxford Nanopore long-read sequencing results for three species of macroalgae to those of non-macroalgal species and find that macroalgal samples tend to lead to rapid decline in the number of available sequencing pores resulting in reduced sequencing yield. LC-MS analysis of macroalgal DNA that would be considered suitable for sequencing reveals that DNA derived from dried macroalgae is enriched for polyphenol-DNA adducts - which may lead to sequencing inhibition. Our findings have wide-ranging implications for the generation of genomic resources from macroalgae, for example long read sequencing of dried herbarium specimens, and suggest a need to use fresh tissue wherever possible for genome sequencing.

molecular biology↗

Spatial Transcriptomics Reveals the Requirement of ADGRG6 in Maintaining Chondrocyte Homeostasis in Mouse Growth Plates

The growth plate is essential for maintaining skeletal growth; however, the mechanisms governing postnatal growth plate homeostasis are poorly understood. Here we show that ADGRG6/GPR126, a cartilage-enriched G protein-coupled receptor (GPCR), is dispensable for embryonic limb development but is required for postnatal growth plate homeostasis. Adgrg6 ablation in osteochondral progenitor cells or postnatal chondrocytes leads to reduced cellularity and impaired maintenance of the resting zone in the growth plate, coupled with increased cell death and reduced cell proliferation. Adgrg6 mutant growth plates also exhibit disorganized extracellular matrix structures and dysregulated hypertrophic differentiation. Furthermore, using a novel spatial transcriptomics workflow that applies to FFPE tissue sections of mineralized mouse knee joints, we demonstrate that Adgrg6 ablation leads to reduced SOX9 expression, induced Indian hedgehog (IHH) signaling, and a precocious chondrogenic-to-osteogenic conversion of the growth plate chondrocytes that may be driven by increased POSTN/integrin receptor signaling. We further demonstrated that ADGRG6 regulates the proper formation of the resting zone growth plate by maintaining the PTHrP and SOX9-positive cell populations. Altogether, our findings elucidate the essential role of ADGRG6 in maintaining chondrocyte fate, survival, and homeostasis of the postnatal growth plates.

molecular biology↗

Mutation in the Bone Morphogenetic Protein signaling pathway sensitize zebrafish and human to ethanol-induced jaw malformations

Fetal Alcohol Spectrum Disorders (FASD) describe ethanol-induced developmental defects including craniofacial malformations. While ethanol-sensitive genetic mutations contribute to facial malformations, the impacted cellular mechanisms remain unknown. Bmp signaling is a key regulator of epithelial morphogenesis driving facial development, providing a possible ethanol-sensitive mechanism. We found that zebrafish mutants for Bmp signaling components are ethanol-sensitive and affect anterior pharyngeal endoderm shape and gene expression, indicating ethanol-induced malformations of the anterior pharyngeal endoderm cause facial malformations. Integrating FASD patient data, we provide the first evidence that variants in the human Bmp receptor gene BMPR1B associate with ethanol-related differences in jaw volume. Our results show that ethanol exposure disrupts proper morphogenesis of, and tissue interactions between, facial epithelia that mirror overall viscerocranial shape changes and are predictive for Bmp-ethanol associations in human jaw development. Our data provide a mechanistic paradigm linking ethanol to disrupted epithelial cell behaviors that underlie facial defects in FASD. Summary StatementIn this study, we apply a unique combination of zebrafish-based approaches and human genetic and facial dysmorphology analyses to resolve the cellular mechanisms driven by the ethanol-sensitive Bmp pathway.

developmental biology↗

Whole-genome sequencing confirms multiple species of Galapagos giant tortoises

Galapagos giant tortoises are endemic to the Galapagos Archipelago, where they are found in isolated populations. While these populations are widely considered distinguishable in morphology, behavior, and genetics, the recent divergence of these taxa has made their status as species controversial. Here, we apply multispecies coalescent methods for species delimitation to whole genome resequencing data from 38 tortoises across all 13 extant taxa to assess support for delimiting these taxa as species. In contrast to previous studies based solely on divergence time, we find strong evidence to reject the hypothesis that all Galapagos giant tortoises belong to a single species. Instead, a conservative interpretation of model-based and divergence-based results indicates that these taxa form a species complex consisting of a minimum of 9 species, with some analyses supporting as many as 13 species. There is mixed support for the species status of taxa living on the same island, with some methods delimiting them as separate species and others suggesting multiple populations of a single species per island. These results make clear that Galapagos giant tortoise taxa represent different stages in the process of speciation, with some taxa further along in that evolutionary process than others. A better understanding of the more complex parts of that process is urgently needed, given the threatened status of Galapagos giant tortoises. Lay SummarySpecies delimitation is a challenging problem in evolutionary biology, but one that is central to the field. Distinguishing species can affect conservation management practices, from conservation status assessments to strategies for breeding programs. More fundamentally, understanding species boundaries affects our ability to assess biodiversity and to study evolutionary processes. The Galapagos Archipelago presents several radiations of closely related taxa that inspired Charles Darwin to develop his theory of evolution by natural selection and later led to foundational case studies in speciation. The Galapagos giant tortoises were one such inspiration. Nearly two centuries later, there is still an ongoing debate about the taxonomic status of these tortoises, with opinions on their status ranging from barely differentiated populations to separate species. Here, we present the first genomic species delimitation of Galapagos giant tortoises and provide convincing evidence that this group is a complex consisting of between 9 and 13 species. These results provide valuable guidance to conservation stakeholders in the Galapagos, while also adding an important case study to the delimitation of island species.

evolutionary biology↗

Kif6 regulates cilia motility and polarity in brain ependymal cells

Ependymal cells, lining brain ventricular walls, display tufts of cilia that beat in concert promoting laminar Cerebrospinal fluid (CSF) flow within brain ventricles. The ciliary axonemes of multiciliated ependymal cells display a 9+2 microtubule array common to motile cilia. Dyneins and kinesins are ATPase microtubule motor proteins that promote the rhythmic beating of cilia axonemes. Despite common consensus about the importance of axonemal dynein motor proteins, little is known about how Kinesin motors contribute to cilia motility. Here, we define the function of Kinesin family member 6 (Kif6) using a mutation that lacks a highly conserved C-terminal tail domain (Kif6p.G555fs) and which displays progressive hydrocephalus in mice. An analogous mutation was isolated in a proband displaying macrocephaly, hypotonia, and seizures implicating an evolutionarily conserved function for Kif6 in neurodevelopment. We find that loss of Kif6 function caused decreased ependymal cilia motility and subsequently decreased fluid flow on the surface of brain ventricular walls. Kif6 protein was localized at ependymal cilia and displayed processive motor movement (676 nm/s) on microtubules in vitro. Loss of the Kif6 C-terminal tail domain did not affect the initial ciliogenesis in vivo, but did result in defects in cilia orientation, the formation of robust apical actin networks, and stabilization of basal bodies at the apical surface. This suggests a novel role for the Kif6 motor in maintenance of ciliary homeostasis of ependymal cells. Summary statementWe found that Kif6 is localized to the axonemes of ependymal cells. In vitro analysis shows that Kif6 moves on microtubules and that its loss mice decrease cilia motility and cilia-driven flow, resulting in hydrocephalus.

developmental biology↗

Does ivermectin treatment for endemic hookworm infection alter the gut microbiota of endangered Australian sea lion pups?

The gut microbiota is essential for the development and maintenance of the hosts immune system, and disturbances can impact host health. This study aimed to determine if topical ivermectin treatment for endemic hookworm (Uncinaria sanguinis) infection in Australian sea lion (Neophoca cinerea) pups causes gut microbial changes. The gut microbiota was characterised for untreated (control) (n=23) and treated (n=23) pups sampled during the 2019 and 2020/21 breeding seasons at Seal Bay, Kangaroo Island. Samples were collected pre- and post-treatment on up to four occasions. The gut microbiota of both untreated (control) and treated pups was dominated by five bacterial phyla, Fusobacteria, Firmicutes, Proteobacteria, Actinobacteria and Bacteroides. There was a significant difference in alpha diversity between treatment groups in 2020/21 (p = 0.008), with greater diversity in treated pups. Modelling the impact of host factors on beta diversity revealed that pup ID accounted for most of the variation with pup ID, age and capture being the only significant contributors to microbial variation (p < 0.05). There were no statistically significant differences in microbial composition between treatment groups in both breeding seasons, indicating that ivermectin treatment did not alter microbial composition. To our knowledge, this is the first study to consider the impact of parasitic treatment on overall diversity and composition of the gut microbiota. Importantly, the lack of compositional changes in the gut microbiota with topical treatment support the utility of topical ivermectin as a safe and minimally invasive management strategy to enhance pup survival in this endangered species. ImportanceDisturbances to the gut microbiota in early life stages can have life-long impacts on host health. Australian sea lions are endangered and declining, and pups are endemically infected with hookworm (Uncinaria sanguinis) which contributes to pup mortality. Treatment with topical ivermectin has been shown to effectively eliminate hookworm infection and to improve pup health, but the impact on the gut microbiota was previously unknown, representing a key knowledge gap. The results from this study show that topical ivermectin treatment does not alter the gut microbiota of Australian sea lion pups, indicating that it is a safe and minimally invasive treatment that can aid in disease mitigation and conservation of this endangered species.

microbiology↗

Swarm learning for decentralized artificial intelligence in cancer histopathology

Artificial Intelligence (AI) can extract clinically actionable information from medical image data. In cancer histopathology, AI can be used to predict the presence of molecular alterations directly from routine histopathology slides. However, training robust AI systems requires large datasets whose collection faces practical, ethical and legal obstacles. These obstacles could be overcome with swarm learning (SL) where partners jointly train AI models, while avoiding data transfer and monopolistic data governance. Here, for the first time, we demonstrate the successful use of SL in large, multicentric datasets of gigapixel histopathology images comprising over 5000 patients. We show that AI models trained using Swarm Learning can predict BRAF mutational status and microsatellite instability (MSI) directly from hematoxylin and eosin (H&E)-stained pathology slides of colorectal cancer (CRC). We trained AI models on three patient cohorts from Northern Ireland, Germany and the United States of America and validated the prediction performance in two independent datasets from the United Kingdom using SL-based AI models. Our data show that SL enables us to train AI models which outperform most locally trained models and perform on par with models which are centrally trained on the merged datasets. In addition, we show that SL-based AI models are data efficient and maintain a robust performance even if only subsets of local datasets are used for training. In the future, SL can be used to train distributed AI models for any histopathology image analysis tasks, overcoming the need for data transfer and without requiring institutions to give up control of the final AI model.

bioinformatics↗

Carriage of antibiotic resistant bacteria in endangered and declining Australian pinniped pups

The rapid emergence of antimicrobial resistance (AMR) is a major concern for wildlife and ecosystem health globally. Genetic determinants of AMR have become indicators of anthropogenic pollution due to their greater association with humans and rarer presence in environments less affected by humans. The objective of this study was to determine the distribution and frequency of the class 1 integron, a genetic determinant of AMR, in both the faecal microbiome and in Escherichia coli isolated from neonates of three pinniped species. Australian sea lion (Neophoca cinerea), Australian fur seal (Arctocephalus pusillus doriferus) and long-nosed fur seal (Arctocephalus forsteri) pups from eight breeding colonies along the Southern Australian coast were sampled between 2016-2019. DNA from faecal samples (n=309) and from E. coli (n=795) isolated from 884 faecal samples were analysed for class 1 integrons using PCRs targeting the conserved integrase gene (intI) and the gene cassette array. Class 1 integrons were detected in A. p. doriferus and N. cinerea pups sampled at seven of the eight breeding colonies investigated in 4.85% of faecal samples (n=15) and 4.52% of E. coli isolates (n=36). Integrons were not detected in any A. forsteri samples. DNA sequencing of the class 1 integron gene cassette array identified diverse genes conferring resistance to four antibiotic classes. The relationship between class 1 integron carriage and the concentration of five trace elements and heavy metals was also investigated, finding no significant association. The results of this study add to the growing evidence of the extent to which antimicrobial resistant bacteria are polluting the marine environment. As AMR determinants are frequently associated with bacterial pathogens, their occurrence suggests that these pinniped species are vulnerable to potential health risks. The implications for individual and population health as a consequence of AMR carriage is a critical component of ongoing health investigations.

microbiology↗

TopoStats - an automated tracing program for AFM images

We present TopoStats, a Python toolkit for automated editing and analysis of Atomic Force Microscopy images. The program automates identification and tracing of individual molecules in circular and linear conformations without user input. TopoStats was able to identify and trace a range of molecules within AFM images, finding, on average, 90% of all individual molecules and molecular assemblies within a wide field of view, and without the need for prior processing. DNA minicircles of varying size, DNA origami rings and pore forming proteins were identified and accurately traced with contour lengths of traces typically within 10 nm of the predicted contour length. TopoStats was also able to reliably identify and trace linear and enclosed circular molecules within a mixed population. The program is freely available via GitHub (https://github.com/afm-spm/TopoStats) and is intended to be modified and adapted for use if required.

biophysics↗

Genomic characterization of the adolescent idiopathic scoliosis associated transcriptome and regulome

Adolescent idiopathic scoliosis (AIS), a sideways curvature of the spine, is the most common pediatric musculoskeletal disorder, affecting [~]3% of the population worldwide. However, its genetic bases and tissues of origin remain largely unknown. Several genome-wide association studies (GWAS) have implicated nucleotide variants in noncoding sequences that control genes with important roles in cartilage, muscle, bone, connective tissue and intervertebral discs (IVDs) as drivers of AIS susceptibility. Here, we set out to define the expression of AIS-associated genes and active regulatory elements by performing RNA-seq and ChIP-seq against H3K27ac in these tissues in mouse and human. Our study highlights genetic pathways involving AIS-associated loci that regulate chondrogenesis, IVD development and connective tissue maintenance and homeostasis. In addition, we identify thousands of putative AIS-associated regulatory elements which may orchestrate tissue-specific expression in musculoskeletal tissues of the spine. Quantification of enhancer activity of several candidate regulatory elements from our study identifies three functional enhancers carrying AIS-associated GWAS SNPs at the ADGRG6 and BNC2 loci. Our findings provide a novel genome-wide catalog of AIS-relevant genes and regulatory elements and aid in the identification of novel targets for AIS causality and treatment.

genetics↗

The genomic landscape of western South America: Andes, Amazonia and Pacific Coast

Studies of Native South American genetic diversity have helped to shed light on the peopling and differentiation of the continent, but available data are sparse for the major ecogeographic domains. These include the Pacific Coast, a potential early migration route; the Andes, home to the most expansive complex societies and to one of the most spoken indigenous language families of the continent (Quechua); and Amazonia, with its understudied population structure and rich cultural diversity. Here we explore the genetic structure of 177 individuals from these three domains, genotyped with the Affymetrix Human Origins array. We infer multiple sources of ancestry within the Native American ancestry component; one with clear predominance on the Coast and in the Andes, and at least two distinct substrates in neighboring Amazonia, with a previously undetected ancestry characteristic of northern Ecuador and Colombia. Amazonian populations are also involved in recent gene-flow with each other and across ecogeographic domains, which does not accord with the traditional view of small, isolated groups. Long distance genetic connections between speakers of the same language family suggest that languages had spread not by cultural contact alone. Finally, Native American populations admixed with post-Columbian European and African sources at different times, with few cases of prolonged isolation. With our results we emphasize the importance of including under-studied regions of the continent in high-resolution genetic studies, and we illustrate the potential of SNP chip arrays for informative regional scale analysis.

genetics↗

Nanoscale Polarization of the Vaccinia Virus Entry Fusion Complex Drives Efficient Fusion

The spatial distribution of binding and fusion proteins on most viruses and the functional relevance of this organization remains largely unexplored. Employing super-resolution microscopy we define the nanoscale membrane architecture of the prototypic poxvirus, vaccinia. We show that binding and entry fusion complex (EFC) proteins are organized into distinct functional domains with fusion proteins polarized to the tips of virions. Repression of individual EFC components disrupted fusion protein polarization, correlating with a loss of fusion activity. Repression of vaccinia A27, a non-EFC protein implicated in fusion, revealed that disruption of EFC localization impacts virus fusion pore formation. We propose that the polarized distribution of EFCs is essential for poxvirus fusion efficiency.

microbiology↗

Mutations in Kinesin Family Member 6 Reveal Specific Role in Ependymal Cell Function and Human Neuro-Cranial Development

Cerebrospinal fluid flow is crucial for neurodevelopment and homeostasis of the ventricular system of the brain, with localized flow being established by the polarized beating of the ependymal cell (EC) cilia. Here, we report a homozygous one base-pair deletion, c.1193delT (p.Leu398Glnfs*2), in the Kinesin Family Member 6 (KIF6) gene in a child displaying neurocranial defects and intellectual disability. To test the pathogenicity of this novel human KIF6 mutation we engineered an analogous C-terminal truncating mutation in mouse. These mutant mice display severe, postnatal-onset hydrocephalus. We generated a Kif6-LacZ transgenic mouse strain and report expression specifically and uniquely within the ependymal cell (EC) layer of the brain, without labeling other multiciliated mouse tissues. Analysis of Kif6 mutant mice with scanning electron microscopy (SEM) and immunofluorescence (IF) revealed a reduction in EC cilia, without effect on other multiciliated tissues. Consistent with our findings in mice, defects of the ventricular system and EC cilia were observed in kif6 mutant zebrafish. Overall, this work describes the first clinically-defined KIF6 homozygous null mutation in human and defines KIF6 as a conserved mediator of neuro-cranial morphogenesis with a specific role in the maintenance of EC cilia in vertebrates.\n\nAUTHOR SUMMARYCerebrospinal fluid flow is crucial for neurodevelopment and homeostasis of the ventricular system of the brain. Localized flows of cerebrospinal fluid throughout the ventricular system of the brain are established from the polarized beating of the ependymal cell (EC) cilia. Here, we identified a homozygous truncating mutation in KIF6 in a child displaying neuro-cranial defects and intellectual disability. To test the function of KIF6 in vivo, we engineered mutations of Kif6 in mouse. These Kif6 mutant mice display severe hydrocephalus, coupled with a loss of EC cilia. Similarly, we observed hydrocephalus and a reduction in EC cilia in kif6 mutant zebrafish. Overall, this work describes the first clinically-defined KIF6 mutation in human, while our animal studies demonstrate the pathogenicity of mutations in KIF6 and establish KIF6 as a conserved mediator of neuro-cranial development and EC cilia maintenance in vertebrates.\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=183 SRC=\"FIGDIR/small/350140_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (58K):\norg.highwire.dtl.DTLVardef@d36bc6org.highwire.dtl.DTLVardef@48cbfborg.highwire.dtl.DTLVardef@147e273org.highwire.dtl.DTLVardef@641ce3_HPS_FORMAT_FIGEXP M_FIG C_FIG

genetics↗