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Biology subjects

Davey, M. G.

Publications and source records attributed to Davey, M. G..

8 recordsLinked to original sources

A frameshift mutation in PRICKLE1 causes skeletal dysplasia in Cheviot sheep

Skeletal dysplasias can occur in animals, including humans, and are attributed to both environmental and genetic causes. As our understanding of the distinguishing phenotypes of different skeletal dysplasias improves, alongside elucidation of the mechanisms and genetic networks that contribute to skeletal development, identifying and controlling genetic mutations in livestock populations has become increasingly possible, including approaches beyond selective breeding alone. Within Scotland an 'achondroplasia' syndrome that was first described in the South Country Cheviot breed of sheep (OMIA 000004-9940), has continued to occur periodically within the same geographical locations, despite culling of affected lambs over many decades. Affected lambs exhibit rounded heads, short legs, a loss of hooves, abnormal stance and short twisted tails. As the condition has been observed in flocks and as progeny from specific ram and ewe matings, and cases have been co-twinned to normal siblings, this suggests a syndrome caused by a recessive genetic condition rather than environmental factors. Through homozygosity mapping and whole genome sequence analyses, we identified a 10 bp deletion in the open reading frame of the PRICKLE1 gene that encodes prickle planar cell polarity protein 1 and is essential for normal vertebrate embryonic development. We identified additional phenotypes present in affected lambs, including a loss of phalanges, neural tube defects, vertebral and articular chondrocyte defects, hair orientation differences and polycystic kidneys, consistent with a loss of PRICKLE1 function based on comparisons to mouse mutant lines. These findings enabled the development of a simple DNA test for the identified PRICKLE1 mutation that was made commercially available to facilitate the elimination of this genetic disease from sheep flocks.

genetics↗

FuChi: A cell cycle biosensor for investigating cell-cycle kinetics during avian development.

The ability to monitor the proliferative status of live cells both in vitro and in vivo over time has revolutionised our understanding of development, growth and disease. This was first made possible by fluorescent ubiquitination-based cell cycle indicator (Fucci) technology, which distinguishes specific cell cycle phases through the reciprocal degradation of fluorescently tagged, truncated forms of human CDT1 and GMNN proteins. Fucci genetic systems have been successfully implemented in transgenic mice, zebrafish, and axolotls. To date, no viable, stably expressing Fucci line has been developed in an avian species. Although a range of continuously improving Fucci constructs have been developed in recent years, existing in vivo Fucci models remain limited because they rely on older reporter technology that fails to distinguish cells in S, G2, and M phases or to label cells in early G1. As a result, these models can be challenging to interpret and their utility for continuous cell tracking and precise analysis of cell-cycle dynamics is limited. Here, we introduce FuChi, a multicistronic Fucci-expressing chicken line incorporating a newly optimised reporter construct composed of an mCerulean-tagged Histone H1.0 linker protein fused via a self-cleaving 2A peptide to the tandem Fucci(CA) cell cycle biosensor, with additional epitope tags included for detection in fixed tissues. We show that this system accurately discriminates and permits tracking of cells in G1, S, G2, and M phases both in vitro and in vivo, enabling faithful visualisation of cell cycle status in intact tissues and organs. Using FuChi embryos, we mapped proliferation dynamics across developing tissues, analysed cell cycle states of migrating cells, and performed live imaging of early embryos. These latter experiments revealed that transition from S phase may be a key morphogenetic event during gastrulation as mesendoderm cells egress from the primitive streak to form embryonic structures including the prechordal plate. Pairing this advanced reporter with the intrinsic experimental advantages of the chicken embryo positions FuChi as a premier in vivo system for studying cell-cycle kinetics in development, delivering clear technological improvements over current Fucci models. FuChi chickens provide a powerful new resource for studying embryonic development, organ growth, tissue homeostasis, disease processes, and infection responses.

developmental biology↗

Characterisation of the Avascular Mesenchyme during Digit Outgrowth

The avascular mesenchyme at the tip of the developing digit contributes to digit outgrowth and patterning, however, it has been poorly characterised. Using newly developed fate mapping approaches, tissue manipulation and single-cell mRNA sequencing data, we explore the transcriptional nature and developmental potential of this tissue. We find that the avascular mesenchyme is essential to normal segmental patterning of the digit and has a distinct transcriptional identity. In addition, we uncover an unexpected relationship between the unspecified tissue of the avascular mesenchyme and the committed phalanx forming region, which controls patterning, but not outgrowth of the digit. This multifaceted approach provides insights into the mechanics and genetic pathways that regulate digit outgrowth and patterning. HighlightsO_LIThe Avascular Mesenchyme of the developing digit contains a population of uncommitted cells, with a unique transcriptional identity. C_LIO_LIThe AVM is critical to the normal outgrowth and patterning of digits. C_LIO_LIHMGA1 expression is associated with the Avascular Mesenchyme C_LI

developmental biology↗

A scientific case for revisiting the embryonic chicken model in biomedical research

The availability of fertilised chicken eggs and the accessibility and rapid development of the avian embryo, have been utilised in biomedical scientific research to make fundamental discoveries including of developmental processes that are common to all vertebrates, advances in teratology, the understanding of tumour growth and metastasis, angiogenesis, cancer drug assessment and vaccine development as well as advances in understanding avian specific biology. However, recent innovations in chicken transgenesis, genome engineering and surrogate host technology in chickens have only been utilised in a few of these fields of research, specifically some areas of developmental biology, avian sex determination and immunology. To understand why other biomedical fields have not adopted modern chicken transgenic tools, we investigated the non-technical summaries of projects granted in the UK under the Animals (Scientific Procedures) Act 1986 between 2017-2023 to assess when and how chicken embryos are used in research, and if they were considered as a Replacement model for other species. HighlightsO_LIEvidence for a lack of uptake of the chicken embryo as a partial replacement model C_LIO_LICase examples of the reduction in animal numbers when chicken models are used C_LIO_LIProposed action plan for the avian developmental biology community C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC="FIGDIR/small/623941v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@a78644org.highwire.dtl.DTLVardef@6007d4org.highwire.dtl.DTLVardef@a21fb3org.highwire.dtl.DTLVardef@1cf5b90_HPS_FORMAT_FIGEXP M_FIG C_FIG

developmental biology↗

Insights into Digit Evolution from a Fate Map Study of the Forearm

The cellular and genetic networks which contribute to the development of the zeugopod, (radius and ulna of the forearm, tibia and fibula of the leg) are not well understood, although these bones are susceptible to loss in congenital human syndromes and to the action of teratogens such as thalidomide. Using a new fate mapping approach in transgenic chickens, we show that there is a small contribution of SHH expressing cells to the posterior ulna, posterior carpals and digit 3. We establish that while the majority of the ulna develops in response to paracrine SHH signaling in both the chicken and mouse, there are differences in the contribution of SHH expressing cells to other tissues of the zeugopod between these two species as well as between the chicken ulna and fibula. This is evidence that although zeugopod bones are clearly homologous according to the fossil record, the zeugopod bones of the wing and leg are formed by subtly different signalling and patterning events during embryonic development, which can be used to understand the shaping of the bird wing skeleton during the evolution of powered flight.

developmental biology↗

Injury and a program of fetal wound healing in the fetal and neonatal extrahepatic bile duct

IntroductionBiliary atresia (BA) is an obstructive cholangiopathy that initially affects the extrahepatic bile ducts (EHBDs) of neonates. The etiology is uncertain, but evidence points to a prenatal cause; however, the response of the fetal EHBD to injury remains unknown. The objective of this study was to define the fetal response to EHBD injury and to determine whether it follows a fetal wound healing paradigm. MethodsMouse, rat, sheep, and human EHBD samples were studied at different developmental time points. Models included a fetal sheep model of prenatal hypoxia, human BA EHBD remnants and liver samples taken at the time of the Kasai procedure, EHBDs isolated from neonatal rats and mice, and spheroids and other models generated from primary neonatal mouse cholangiocytes. ResultsA wide layer of high molecular weight HA encircling the lumen was characteristic of the normal perinatal but not adult EHBD. This layer, which was surrounded by collagen, expanded in injured ducts in parallel with extensive peribiliary gland (PBG) hyperplasia, increased mucus production and elevated serum bilirubin levels. BA EHBD remnants similarly showed increased HA centered around ductular structures compared with age-appropriate controls. High molecular weight HA typical of the fetal/neonatal ducts caused increased cholangiocyte spheroid growth, whereas low molecular weight HA induced abnormal epithelial morphology; low molecular weight HA caused matrix swelling in a bile duct-on-a-chip device. ConclusionThe fetal/neonatal EHBD, including in human EHBD remnants from Kasai surgeries, demonstrated an injury response with high levels of HA typical of the regenerative, scarless program termed fetal wound healing. Although generally beneficial, the expanded peri-luminal HA layer may swell and lead to elevated bilirubin levels and obstruction of the EHBD.

physiology↗

TALPID3/KIAA0586 regulates multiple aspects of neuromuscular patterning during gastrointestinal development in animal models and human

TALPID3/KIAA0586 is an evolutionary conserved protein, which plays an essential role in protein trafficking. Its role during gastrointestinal (GI) and enteric nervous system (ENS) development has not been studied previously. Here, we analysed chicken, mouse and human embryonic GI tissues with TALPID3 mutations. The GI tract of TALPID3 chicken embryos was shortened and malformed. Histologically, the gut smooth muscle was mispatterned and enteric neural crest cells were scattered throughout the gut wall. Analysis of the Hedgehog pathway and gut extracellular matrix provided causative reasons for these defects. Interestingly, chicken intra-species grafting experiments and a conditional knockout mouse model showed that ENS formation did not require TALPID3, but was dependent on correct environmental cues. Surprisingly, the lack of TALPID3 in enteric neural crest cells (ENCC) affected smooth muscle and epithelial development in a non cell-autonomous manner. Analysis of human gut fetal tissues with a KIAA0586 mutation showed strikingly similar findings compared to the animal models demonstrating conservation of TALPID3 and its necessary role in human GI tract development and patterning

developmental biology↗

The transcriptional signature associated with human motile cilia

Cilia are complex microtubule-based organelles implicated in the aetiology of numerous diseases. Accordingly, many cilia-associated proteins have been described, while those distinguishing cilia subtypes are poorly defined. Here, we characterise the gene signature associated with human motile cilia that captures both known and unknown components of this class of cilia. To define the signature, we performed network deconvolution of transcriptomics data derived from tissues possessing motile ciliated cell populations. For each tissue, genes coexpressed with the motile cilia-associated transcriptional factor, FOXJ1, were identified. The consensus across tissues provided a transcriptional signature of 248 genes. For validation, we examined the literature, databases, single cell RNA-Seq data, and the localisation of mRNA and proteins in motile ciliated cells. To validate some of the many poorly characterised genes, we performed new localisation experiments on ARMC3, EFCAB6, FAM183A, MYCBPAP, RIBC2 and VWA3A. In summary, we report a highly validated set of motile cilia-associated genes that helps shape our understanding of these complex cellular organelles.\n\nSummaryThis work defines a conserved transcriptional signature associated with human motile cilia, including many genes with little or no previous association with these structures. These genes were compared with existing resources and a number of poorly characterised genes validated.\n\nGraphical abstract\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=197 SRC=\"FIGDIR/small/817072v1_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (74K):\norg.highwire.dtl.DTLVardef@152f5d6org.highwire.dtl.DTLVardef@189eba6org.highwire.dtl.DTLVardef@1cde94forg.highwire.dtl.DTLVardef@1906450_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗