Biology subjects

Amano, T.

Publications and source records attributed to Amano, T..

18 recordsLinked to original sources

bioRxiv2026

A patient-derived LMX1B variant causes tissue-specific manifestations of nail-patella syndrome in mice

Nail-patella syndrome (NPS) is a multisystem disorder caused by pathogenic variants in LMX1B and is characterized by dysplasia of the nails and patellae as well as extraskeletal complications such as progressive nephropathy and glaucoma. We generated a CRISPR/Cas9 knock-in mouse carrying the R252Q substitution, corresponding to a human LMX1B variant associated with renal-predominant disease. Phenotypic analysis revealed that homozygous mice were viable, but they displayed marked growth retardation and severe bilateral ocular opacity. Interestingly, while this model exhibited clear skeletal and ocular defects, the renal phenotype was relatively mild, although increased urinary albumin excretion, focal glomerular basement membrane abnormalities, and subtle changes in renal gene expression were detected. Beyond the classical NPS hallmarks, mutant mice also displayed midbrain morphological abnormalities, suggesting broader developmental consequences of this LMX1B variant. This patient-derived variant model not only recapitulates the pleiotropic features of NPS but also demonstrates organ-specific susceptibility to the R252Q substitution, providing a foundation for elucidating the complex molecular mechanisms underlying multisystem disease.

genetics↗

bioRxiv2026

EV-Tracer enables lineage-resolved detection and molecular profiling of extracellular vesicle-associated signals in cancer-fibroblast co-culture

Extracellular vesicles (EVs) facilitate intercellular communication by transferring diverse bioactive molecules from donor to recipient cells. However, EVs released by distinct cellular lineages become difficult to distinguish when mixed in multicellular experimental models, limiting the analysis of how cell-cell interactions affect EV-associated molecular profiles. To address this, EV-Tracer, a CD63-based dual-fluorescence tracing and capture system for detecting, isolating, and profiling lineage-associated EV fractions, was developed. Achilles or mScarlet was inserted into the small extracellular loop of CD63, enabling tracer-specific EV detection by digital counting, antibody-based isolation, and live-cell visualisation. Exploratory EV RNA sequencing suggested that physical cell-cell contact was associated with distinct EV RNA profiles, including interferon-related signals, which were supported by targeted cellular and EV-associated RNA analyses. EV-Tracer provides a practical framework for investigating lineage-associated EV dynamics and molecular signals in mixed-cell systems.

cell biology↗

bioRxiv2026

What we miss shapes what we protect: sea turtles as a case study of grey literature and language bias in an evidence synthesis

Evidence-based approaches are increasingly advocated and applied in conservation to provide reliable recommendations for management and policymaking. Although substantial efforts have been undertaken to generate ecological knowledge across languages and publication types, many syntheses and meta-analyses fail to include relevant literature sources; drawing into question their reliability. To assess the extent of biases resulting from ignoring grey literature and non-English publications, we reviewed and synthesised movement and migratory connectivity data for green turtles across Southeast Asia from multiple literature types and eight languages. We identified substantial losses in connectivity information, with at least half of all locations losing more than 88% of their connections, and shifts in demographic representation and sampling techniques when grey literature was excluded from the evidence base. The exclusion of non-English literature had a comparatively smaller impact. The regional connectivity network generated through this study highlights the need for holistic reviews to develop evidence bases to underpin transboundary management of migratory species. Ignoring grey literature and, potentially, regional languages in the development may lead to misallocation of limited management resources. Holistic reviews can further support the establishment of Important Areas for migratory species and inform how we meet and report on global biodiversity commitments.

ecology↗

bioRxiv2026

Oxytocin regulates TN-GnRH3 circuit maturation and mate preference through C1q-dependent synaptic mechanisms

Oxytocin is a key regulator of social behavior, yet how it shapes the neural circuits underlying these behaviors remains unclear. Here, we show that oxytocin signaling is required for the proper maturation of terminal nerve gonadotropin-releasing hormone 3 (TN-GnRH3) circuits that regulate female mate preference in medaka. Disruption of oxytocin signaling reduced expression of the complement component C1q, altered microglial properties, and increased TN-GnRH3 innervation in the optic tectum and dorsolateral telencephalon, suggesting impaired synaptic refinement. In addition, TN-GnRH3 neurons in mutants exhibited disrupted clustering and failed to increase firing frequency in response to visual stimulation, indicating functional deficits. Furthermore, c1qb mutants phenocopied the behavioral and neuronal abnormalities observed in oxytocin signaling-deficient mutants. Together, our findings suggest that oxytocin signaling links microglia-associated synaptic refinement to neural circuit maturation, thereby revealing a previously unrecognized role for oxytocin in shaping social behavior.

neuroscience↗

bioRxiv2025

Comprehensive and accelerated mapping of driver mutations through single-nucleotide random mutagenesis of target genes

Identification of novel driver mutations is crucial for personalized medicine and drug discovery. However, genome-wide studies based solely on patient-derived data cannot confirm the oncogenic potential of detected mutations. Here, we present a genome editing-based method for comprehensive random mutagenesis of target genes. This approach generates cultured cells harboring random single-nucleotide variants (SNVs) within 10 days of DNA amplicon preparation. To validate its utility, random mutations were introduced into endogenous exons of lung cancer-related oncogenes in HEK293 cells. Spheroid formation assays and xenograft models demonstrated enhanced tumorigenicity, indicating the acquisition of oncogenic potential. Amplicon-sequencing of spheroid transcripts revealed significant enrichment of variants annotated as pathogenic in clinical databases and computational predictions. Furthermore, osimertinib treatment of lung cancer cell lines with random SNVs on EGFR identified 58 novel resistance mutations. These results highlight the effectiveness of our mutagenesis platform, offering a powerful tool to advance cancer research and precision medicine.

bioengineering↗

bioRxiv2024

Agricultural landscapes with high compositional heterogeneity support both forest and farmland birds in Taiwan

ContextPromoting heterogeneous agricultural landscapes could help to reduce the negative impacts of habitat conversion on biota. However, the benefits of landscape heterogeneity can vary among spatial scales and taxa. ObjectivesTo design biodiversity-friendly landscapes, we use nationwide bird survey data and land use maps to examine the effects of compositional heterogeneity, configurational heterogeneity, and habitat amount at different scales on the species richness of different bird groups. MethodsWe examine the effects of configurational heterogeneity (measured using edge density), compositional heterogeneity (Shannons diversity index of habitat types), and habitat amount (proportion of forest and farmland cover) at both transect (local) and landscape (0.5, 1, or 2 km) scales on the species richness of all breeding birds, forest birds, farmland birds, and introduced birds. ResultsTotal species richness had a hump-shaped relationship with local forest cover, and with farmland cover at landscape scale. Richness of both forest birds and richness of farmland birds increased with Shannons diversity index of habitat types at both local and landscape scales, but only increased with the amount of their preferred habitat at the local scale. Richness of introduced birds was greater in landscapes with higher edge density, suggesting those species are associated with human-dominated landscapes. ConclusionsHigh compositional heterogeneity with low configurational heterogeneity at the landscape scale may help maintain native bird richness while minimising the spread of introduced species in Taiwan. These results can help guide land use planning to achieving biodiversity goals in a country with intensive land-use competition.

ecology↗

bioRxiv2022

The shh limb enhancer is activated in patterned limb regeneration but not in hypomorphic limb regeneration in Xenopus laevis

Xenopus young tadpoles regenerate a limb with the anteroposterior (AP) pattern, but metamorphosed froglets regenerate a hypomorphic limb after amputation. The key gene for AP patterning, shh, is expressed in a regenerating limb of the tadpole but not in that of the froglet. Genomic DNA in the shh limb-specific enhancer, MFCS1 (ZRS), is hypermethylated in froglets but hypomethylated in tadpoles: shh expression may be controlled by epigenetic regulation of MFCS1. Is MFCS1 specifically activated for regenerating the AP-patterned limb? We generated transgenic Xenopus laevis lines that visualize the MFCS1 enhancer activity with a GFP reporter. The transgenic tadpoles showed GFP expression in hoxd13- and shh-expressing domains of developing and regenerating limbs, whereas the froglets showed no GFP expression in the regenerating limbs despite having hoxd13 expression. Genome sequence analysis and co-transfection assays using cultured cells revealed that Hoxd13 can activate Xenopus MFCS1. These results suggest that MFCS1 activation correlates with regeneration of AP-patterned limbs and that re-activation of epigenetically inactivated MFCS1 would be crucial to confer the ability to non-regenerative animals for regenerating a properly patterned limb.

developmental biology↗

bioRxiv2022

Controllable self-replicating RNA vaccine delivered intradermally elicits predominantly cellular immunity

Intradermal delivery of self-replicating RNA (srRNA) is a promising vaccine platform. Considering that human skin temperature is around 33{degrees}C, lower than core body temperature of 37{degrees}C, we have developed an srRNA that functions optimally at skin temperature and is inactivated at or above 37{degrees}C as a safety switch. This temperature-controllable srRNA (c-srRNA), when tested as an intradermal vaccine against SARS-CoV-2, functions when injected naked without lipid nanoparticles. Unlike most currently available vaccines, c-srRNA vaccines predominantly elicit cellular immunity with little or no antibody production. Interestingly, c-srRNA-vaccinated mice produced antigen-specific antibodies upon subsequent stimulation with antigen protein. Antigen-specific antibodies were also produced when B-cell stimulation using antigen protein was followed by c-srRNA booster vaccination. Using c-srRNA, we have designed a pan-coronavirus booster vaccine that incorporates both spike receptor binding domains as viral surface proteins and evolutionarily conserved nucleoproteins as viral non-surface proteins, from both SARS-CoV-2 and MERS-CoV. It can thereby potentially immunize against SARS-CoV-2, SARS-CoV, MERS-CoV, and their variants. c-srRNA may provide a route to activate cellular immunity against a wide variety of pathogens.

immunology↗

bioRxiv2021

Tapping into non-English-language science for the conservation of global biodiversity

The widely held assumption that any important scientific information would be available in English underlies the underuse of non-English-language science across disciplines. However, non-English-language science is expected to bring unique and valuable scientific information, especially in disciplines where the evidence is patchy, and for emergent issues where synthesising available evidence is an urgent challenge. Yet such contribution of non-English-language science to scientific communities and the application of science is rarely quantified. Here we show that non-English-language studies provide crucial evidence for informing global biodiversity conservation. By screening 419,680 peer-reviewed papers in 16 languages, we identified 1,234 non-English-language studies providing evidence on the effectiveness of biodiversity conservation interventions, compared to 4,412 English-language studies identified with the same criteria. Relevant non-English-language studies are being published at an increasing rate, and can expand the geographical (by 12-25%) and taxonomic (by 5-32%) coverage of English-language evidence, especially in biodiverse regions, albeit often based on less robust study designs. Our results show that synthesising non-English-language studies is key to overcoming the widespread lack of local, context-dependent evidence and facilitating evidence-based conservation globally. We urge wider disciplines to rigorously reassess the untapped potential of non-English-language science in informing decisions to address other global challenges.

scientific communication and education↗

bioRxiv2021

Language barriers in global bird conservation

Multiple languages being spoken within a species distribution can impede communication among conservation stakeholders, the compilation of scientific information, and the development of effective conservation actions. Here, we investigate the number of official languages spoken within the distributions of 10,863 bird species to identify which ones might be particularly affected by consequences of language barriers. We show that 1587 species have 10 languages or more spoken within their distributions. Threatened, migratory and wide-ranging species have especially many languages spoken within their distribution. Particularly high numbers of species with many languages within their distribution are found in Eastern Europe, Russia and central and western Asia. Global conservation efforts would benefit from implementing guidelines to overcome language barriers, especially in regions with high species and language diversity.

ecology↗

bioRxiv2020

Poor availability of context-specific evidence hampers decision-making in conservation

Evidence-based conservation relies on robust and relevant evidence. Practitioners often prefer locally relevant studies whose results are more likely to be transferable to the context of planned conservation interventions. To quantify the availability of relevant evidence for amphibian and bird conservation we reviewed Conservation Evidence, a database of quantitative tests of conservation interventions. Studies were geographically clustered and found at extremely low densities - fewer than one study was present within a 2,000 km radius of a given location. The availability of relevant evidence was extremely low when we restricted studies to those studying biomes or taxonomic orders containing high percentages of threatened species, compared to the most frequently studied biomes and taxonomic orders. Further constraining the evidence by study design showed that only 17-20% of amphibian and bird studies used robust designs. Our results highlight the paucity of evidence on the effectiveness of conservation interventions, and the disparity in evidence for local contexts that are frequently studied and those where conservation needs are greatest. Addressing the serious global shortfall in context-specific evidence requires a step change in the frequency of testing conservation interventions, greater use of robust study designs and standardized metrics, and methodological advances to analyze patchy evidence bases.

ecology↗

bioRxiv2019

Insights from two decades of the Student Conference on Conservation Science

Conservation science is a crisis-oriented discipline focused on delivering robust answers to reducing human impacts on nature. To explore how the field might have changed during the past two decades, we analyzed 3,245 applications for oral presentations submitted to the Student Conference on Conservation Science (SCCS) in Cambridge, UK. SCCS has been running every year since 2000, aims for global representation by providing bursaries to early-career conservationists from lower-income countries, and has never had a thematic focus, beyond conservation in the broadest sense. We found that the majority of submissions to SCCS were based on primary biological data collection from local scale field studies in the tropics, contrary to established literature which highlights gaps in tropical research. Our results showed a small increase over time in submissions framed around how nature benefits people as well as a small increase in submissions integrating social science. Our findings also suggest that students and early-career conservationists could provide pathways to increased availability of data from the tropics and for addressing well-known biases in the published literature towards wealthier countries. We hope this research will motivate efforts to support student projects, ensuring data and results are published and made publicly available.

ecology↗

bioRxiv2019

The challenge of heterogeneous evidence in conservation

Conservation efforts to tackle the current biodiversity crisis need to be as efficient and effective as possible. To inform decision-makers of the most effective conservation actions, it is important to identify biases and gaps in the conservation literature to prioritize future evidence generation. We assessed the state of this global literature base using the Conservation Evidence database, a comprehensive collection of quantitative tests of conservation actions (interventions) from the published literature. For amphibians and birds, we investigated the nature of Conservation Evidence spatially and taxonomically, as well as by biome, effectiveness metrics, and study design. Studies were heavily concentrated in Western Europe and North America for birds and particularly amphibians. Studies that used the most robust study designs - Before-After Control-Impact and Randomized Controlled Trials - were also the most geographically restricted. Furthermore, there was no relationship between the number of studies in each 1x1 degree grid cell and the number of species, threatened species or data-deficient species. Taxonomic biases and gaps were apparent for amphibians and birds - some orders were absent from the evidence base and others were poorly represented relative to the proportion of threatened species they contained. Temperate forest and grassland biomes were highly represented, which reinforced observed geographic biases. Various metrics were used to evaluate the effectiveness of a given conservation action, potentially making studies less directly comparable and evidence synthesis more difficult. We also found that the least robust study designs were the most commonly used; studies using robust designs were scarce. Future research should prioritize testing conservation actions on threatened species outside of Western Europe and North America. Standardizing metrics and improving the robustness of study designs used to test conservation actions would also improve the quality of the evidence base for synthesis and decision-making.

ecology↗

bioRxiv2019

Responses of global waterbird populations to climate change vary with latitude

While climate change continues to present a major threat to global biodiversity and ecosystems, most research on climate change impacts do not have the resolution to detect changes in species abundance and are often limited to temperate ecosystems. This limits our understanding of global responses in species abundance--a determinant of ecosystem function and services--to climate change including in the highly-biodiverse tropics. We address this knowledge gap by quantifying abundance responses to climate change in waterbirds, an indicator taxon of wetland biodiversity, at 6,822 sites between -55{degrees} and 64{degrees}. Using 1,303,651 count records since 1990 of 390 species, we show that with temperature increase, the abundance of species and populations decreased at lower latitudes, particularly in the tropics, but increased at higher latitudes. These contrasting responses to temperature increase according to latitude indicate potential global-scale poleward shifts of species abundance under climate change, providing empirical support for predictions by earlier studies. The negative responses to temperature increase in tropical species and populations are of conservation concern, as they are often also threatened by other anthropogenic factors. Our results suggest that existing biases in studies towards temperate regions could underestimate the impact of climate change on waterbirds and other species.

ecology↗

bioRxiv2019

Simple study designs in ecology produce inaccurate estimates of biodiversity responses

O_LIEcologists use a wide range of study designs to estimate the impact of interventions or threats but there are no quantitative comparisons of their accuracy. For example, while it is accepted that simpler designs, such as After (sampling sites post-impact without a control), Before-After (BA) and Control-Impact (CI), are less robust than Randomised Controlled Trials (RCT) and Before-After Control-Impact (BACI) designs, it is not known how much less accurate they are.\nC_LIO_LIWe simulate a step-change response of a population to an environmental impact using empirically-derived estimates of the major parameters. We use five ecological study designs to estimate the effect of this impact and evaluate each one by determining the percentage of simulations in which they accurately estimate the direction and magnitude of the environmental impact. We also simulate different numbers of replicates and assess several accuracy thresholds.\nC_LIO_LIWe demonstrate that BACI designs could be 1.1-1.5 times more accurate than RCTs, 2.9-4.1 times more accurate than BA, 3.8-5.6 times more accurate than CI, and 6.8-10.8 times more accurate than After designs, when estimating to within {+/-}30% of the true effect (depending on the sample size). We also found that increasing sample size substantially increases the accuracy of BACI designs but only increases the precision of simpler designs around a biased estimate; only by using more robust designs can accuracy increase. Modestly increasing replication of both control and impact sites also increased the accuracy of BACI designs more than substantially increasing replicates in just one of these groups.\nC_LIO_LIWe argue that investment into using more robust designs in ecology, where possible, is extremely worthwhile given the inaccuracy of simpler designs, even when using large sample sizes. Based on our results we propose a weighting system that quantitatively ranks the accuracy of studies based on their study design and the number of replicates used. We hope these accuracy weights enable researchers to better account for study design in evidence synthesis when assessing the reliability of a range of studies using a variety of designs.\nC_LI

ecology↗

bioRxiv2019

Vulnerable species interactions are important for the stability of mutualistic networks

Species are central to ecology and conservation. However, it is the interactions between species that generate the functions on which ecosystems and humans depend. Despite the importance of interactions, we lack an understanding of the risk that their loss poses to ecological communities. Here, we quantify risk as a function of the vulnerability (likelihood of loss) and importance (contribution to network stability in terms of species coexistence) of 4330 mutualistic interactions from 41 empirical pollination and seed dispersal networks across six continents. Remarkably, we find that more vulnerable interactions are also more important: the interactions that contribute most to network stability are those that are most likely to be lost. Furthermore, most interactions tend to have more similar vulnerability and importance across networks than expected by chance, suggesting that vulnerability and importance may be intrinsic properties of interactions, rather than only a function of ecological context. These results provide a starting point for prioritising interactions for conservation in species interaction networks and, in areas lacking network data, could allow interaction properties to be inferred from taxonomy alone.

ecology↗

bioRxiv2018

Effects of sampling duration and intervals on confidence in population trend estimates

Species population trends are fundamental to conservation, underpinning lUCN red-list classifications, many national lists of threatened species and are also used globally to convey to policy makers the state of nature. Clearly, its crucial to quantify how much we can trust population trend data. Yet many studies analyzing large numbers of population time series lack a straightforward way to estimate confidence in each trend. Here we artificially degrade 27,930 waterbird population time series to see how often subsets of the data correctly estimate the direction and magnitude of each populations true trend. We find you need to sample many years to be confident that there is no significant trend in a population. Conversely, if a significant trend is detected, even from only a small subset of years, this is likely to be representative of the populations true trend. This means that if a significant decline is detected in a population, it is likely to be correct and conservation action should be taken immediately, but if the trend is insignificant, confidence in this can only be high with many samples. Our full results provide a clear and quantitative way to assign confidence to species trends, and lays the foundation for similar studies of other taxa that can help to add rigor to large-scale population analyses.

ecology↗

bioRxiv2017

Sonic hedgehog is required for neural crest-dependent patterning of the intrinsic tongue musculature

The tongue is a highly specialized muscular organ important for breathing, speech, taste and swallowing. The secreted signaling molecule Sonic hedgehog (Shh) is expressed in dorsal tongue epithelium from the initial developmental stage. In this study, we utilized a series of genetic approaches to investigate the role of Shh during mouse tongue formation. Temporal-specific global deletion of Shh demonstrated a functional requirement for normal patterning of the intrinsic tongue muscles and establishment of the lingual tendon. These defects were reproduced in the mutant with a specific loss of signaling in oropharyngeal epithelium by a Shh cis-enhancer. In these mutants, Ptch1 was lost in the underlying cranial neural crest (CNC)-derived mesenchymal lineage. The importance of Shh was confirmed by generating tissue-specific deletions in the ciliopathy gene Ofd1, which transduces Shh signaling. These results revealed Shh roles in patterning of the mesodermal intrinsic tongue muscles through CNC-derived mesenchyme, including the lingual tendon.

developmental biology↗