Biology subjects

Hughes, K. A.

Publications and source records attributed to Hughes, K. A..

12 recordsLinked to original sources

bioRxiv2026

A nicotine biosensor derived from microbial screening

Physiologically relevant biosensors are in increasingly high demand, yet existing ones are severely limited in the number and type of biomarkers that are detected. The lack of biorecognition elements for most medically relevant biomarkers restricts the development of next generation single and continuous use monitors. Over billions of years, microbes have evolved a vast array of proteins to sense and metabolize small molecules, including those pertinent to human health. Of particular interest to us is the identification and subsequent integration of new microbial redox enzymes into electronic biosensors building off the established electrochemical technology of the continuous glucose monitor. Here we deploy genomic screening to identify analyte specific redox enzymes for biosensor development. As a proof of concept, we report the first electrochemical enzyme-based nicotine biosensor from a novel microbial enzyme, and use a variant with improved catalytic performance to enhance sensor performance. The biosensor detects nicotine over 0.4-100 M, a range relevant to nicotine concentrations present in active smoker sweat, saliva, gastric juice, and urine. This microbial mining approach for discovering redox enzymes expands the sensing parts toolbox available over conventional antibodies and aptamers.

bioengineering↗

bioRxiv2025

Methylphenidate exposure alters brain gene expression and induces transgenerational DNA methylation changes in Poecilia reticulata guppies

Chronic exposure to stimulants is known to affect behavioral phenotypes and epigenetic profiles intergenerationally. We have previously shown that chronic methylphenidate hydrochloride (MPH) exposure in male guppies (Poecilia reticulata) induces persistent, paternally transmitted behavioral changes across multiple unexposed generations. Here, we investigated the underlying epigenetic signatures of this transgenerational behavioral inheritance. Building on our previous study, which used composite behavioral scores, we investigates the transgenerational inheritance patters of individual behaviors and found a robust, male-lineage-dependent increase in swimming across unexposed G2-G4 female offspring. Our molecular analysis showed that that chronic MPH exposure in G1 males alters brain gene expression, with 76 differentially expressed genes including genes with developmental and locomotory functions, and an over-representation of long non-coding RNAs. Furthermore, we found that unexposed G4 descendants from MPH-treated lineages exhibit widespread changes in brain DNA methylation, including genes related to developmental and swimming behavior. Interestingly, we found one lnRNA, LOC103476631, as both differentially expressed in G1 males and differentially methylated in G4 fish, providing a compelling candidate for ncRNA-directed DNA methylation as a mechanism for the observed transgenerational epigenetic inheritance of swimming behavior. Author SummaryDrugs used to treat attention-deficit/hyperactivity disorder, such as methylphenidate, are widely prescribed to children and adolescents during key stages of brain development. We previously showed that exposing male guppies to a low, chronic dose of methylphenidate changed anxiety-like behavior not only in the treated fish but also in several generations of their unexposed descendants. In this study, we asked how such long-lasting behavioral effects might be recorded in the brain. We first confirmed that a simple measure of swimming behavior remains altered in descendants of treated males across multiple generations. We then examined brains from the directly exposed fathers and from great-grand-offspring that were never exposed to the drug. In fathers, we found changes in the activity of genes, including many long non-coding RNAs, a class of genes that has been linked to transgenerational inheritance. In great-grand-offspring, we found significantly altered DNA methylation profiles, an epigenetic mark that has also been associated with transgenerational inheritance. Together, our findings suggest that developmental exposure to a commonly used stimulant can leave a stable molecular memory in the brain that persists across generations.

genetics↗

bioRxiv2025

Wearable sweat nicotine biosensor based on a nicotineoxidoreductase and its natural electron acceptor

The advent of wearable biosensors is empowering clinical and lifestyle decision making. Nicotine is a drug of significant negative impact on public health given the prevalence of smoking and vaping. Yet noninvasive, continuous monitoring of nicotine is challenging due to lack of specific and sensitive biosensing elements. Here, we report a wearable highly sensitive nicotine biosensor and demonstrate on-body deployment of the sensor in a first-in-human study. The biosensor comprises nicotine oxidoreductase (NicA2) with its natural cytochrome c electron acceptor (CycN) as mediator: both proteins play a key role in the nicotine-degrading bacterium, Pseudomonas putida S16. The biosensor detects nicotine over four-orders of magnitude (0.1-100 M) with nanomolar sensitivity (LOD [~]33.6 nM), performs in the physiological pH range (pH 6-9), and is highly selective against common interferants and the major human nicotine metabolite, cotinine. Incorporation of the biosensor in a custom wearable device enables real-time measurement of nicotine from locally induced sweat on volunteer subjects and demonstrates higher analytical accuracy than gold-standard mass spectrometry.

bioengineering↗

bioRxiv2025

Repeated signatures of balancing selection in small and large populations of guppies (Poecilia reticulata)

Balancing selection is a powerful evolutionary force that maintains adaptive genetic and phenotypic diversity. Although methods to detect the footprints of balancing selection in genomic data have advanced, we still lack a clear understanding of how repeatable these signatures appear in wild populations, and how this repeatability is shaped by demographic history and existing genetic variation. The Trinidadian guppy (Poecilia reticulata) provides an ideal model to test the repeatability of balancing selection in the wild as there is strong evidence that negative frequency-dependent selection (NFDS) maintains colour polymorphism. Analysing whole-genome sequencing data from 11 guppy populations (n = 195) with contrasting demographic contexts, we apply scans of balancing selection to explore which genomic regions show evidence of repeatability. We find that populations with small Ne show less genetic repeatability but still exhibit population-specific regions of elevated diversity, implicating independent balancing selection or other evolutionary mechanisms. Despite this, we identify 23 regions with repeated signatures of balancing selection, including a region on LG22 containing genes involved in colour, vision, mate choice, and social behaviour. Investigating the repeatability of balancing selection in small and large populations improves our knowledge of how demographic factors interact with selective processes to shape natural variation.

evolutionary biology↗

bioRxiv2024

Getting Started with Machine Learning for Experimental Biochemists and Other Molecular Scientists

Machine learning (ML) is rapidly gaining traction in many areas of experimental molecular science for elucidating relationships and patterns in large or complex data sets. Historically, ML was largely the preserve of those with specialized training in fields such as statistics or cheminformatics. Increasingly, however, ML methodologies are becoming part of the standard toolkit for experimental scientists across a range of disciplines. Lowering the barrier of entry to these ML techniques, for scientists without a significant background in computer science or statistics, is important to broadening access to these powerful methods. Here we provide detailed, step by step tutorials for performing four ML methods that are particularly useful for applications in biochemistry, cell biology, and drug discovery: hierarchical clustering, Principal Component Analysis (PCA), Partial Least-Squares Discriminant Analysis (PLSDA), and Partial Least-Squares Regression (PLSR). The protocols are written for the widely used software MATLAB, but no prior experience with MATLAB is required to use them. We include an explanation of each step, pitched at a level to be understood by investigators without any prior experience with ML, MATLAB, or any kind of coding. We also highlight the scientific issues pertaining to selecting and scaling the data to be analyzed, and describe controls to test the validity of the results obtained. Throughout, we emphasize the relationship between the scientific question and how to choose data and methods that will allow it to be addressed in a meaningful way. Our aim is to provide a basic introduction that will equip experimental chemical biologists and other chemical and biomedical scientists with the knowledge required to use ML to aid in the design of experiments, the formulation and data-driven testing of hypotheses, and the analysis of experimental data. Basic Protocol 1Clustering Basic Protocol 2Principal Component Analysis (PCA) Basic Protocol 3Partial Least Squares Discriminant Analysis (PLSDA) Basic Protocol 4Partial Least Squares Regression (PLSR)

biochemistry↗

bioRxiv2024

A meta-analysis of the relationship between color and aggression

Color varies in pattern and degree across the tree of life. In animals, genetic variation in color is hypothesized to have pleiotropic effects on a variety of behaviors, due to shared dependence on underlying biochemical pathways. Such pleiotropy can constrain the independent evolution of color and behavior. Although associations between color and behavior have been reported, this relationship has not yet been addressed across a broad taxonomic scale with a formal meta-analysis. We used a phylogenetic meta-analytic approach to examine the relationship between individual variation in aggressive behavior and variation in multiple colors. Seventy studies met our inclusion criteria (vertebrates = 66; invertebrates = 4). After accounting for phylogeny and correcting for publication bias, there was a positive association between measures of aggression and degree or area of coloration (mean = 0.274, 95% CI = (0.041, 0.481)). However, this positive association was not influenced by type of color or by several other variables that we tested. Because the data supports a positive association between aggression and degree or area of coloration, irrespective of whether color is melanin-based, carotenoid-based, or structural, we conclude that this pattern does not strongly support the melanin-pleiotropy hypothesis. The relationship was also unaffected by moderators accounting for individual condition, social rank, or age; thus, the results do not strongly support hypotheses that condition-dependence accounts for relationships between color and aggressive behavior. We propose that the moderate positive correlation between aggression and coloration across Animalia that we observed is underlain by genetic covariation between behavior and color traits that serve as badges of status.

evolutionary biology↗

bioRxiv2021

A large and diverse autosomal haplotype is associated with sex-linked colour polymorphism in the guppy

Colour polymorphism provides a tractable trait that can be harnessed to explore the evolution of sexual selection and sexual conflict. Male colour patterns of the Trinidadian guppy (Poecilia reticulata) are governed by both natural and sexual selection, and are typified by extreme pattern colour variation as a result of negative frequency dependent selection. Since guppy colour patterns are often inherited faithfully from fathers to sons, it has been historically presumed that colour genes are physically linked to sex determining loci as a supergene on the sex chromosome. Yet the actual identity and genomic location of the colour pattern genes has remained elusive. We phenotyped and genotyped four guppy Iso-Y lines, where colour was inherited along the patriline, but backcrossed into the stock population every 2 to 3 generations for 40 generations, thereby homogenising the genome at regions unrelated to colour. Using an unbiased phenotyping method to proportion colour pattern differences between and among the Iso-Y lines, we confirmed that the breeding design was successful in producing four distinct colour patterns. Our analysis of genome resequencing data of the four Iso-Y lines uncovered a surprising genetic architecture for colour pattern polymorphism. Genetic differentiation among Iso-Y lines was repeatedly associated with a large and diverse haplotype ([~]5Mb) on an autosome (LG1), not the sex chromosome (LG12). Moreover, the LG1 haplotype showed elevated linkage disequilibrium and exhibited evidence of sex-specific diversity when we examined whole-genome sequencing data of the natural source population. We hypothesise that colour pattern polymorphism is driven by Y-autosome epistasis, and conclude that predictions of sexual conflict should focus on incorporating the effects of epistasis in understanding complex adaptive architectures.

evolutionary biology↗

bioRxiv2021

On the genetic architecture of rapidly adapting and convergent life history traits in guppies

The genetic basis of traits can shape and constrain how adaptation proceeds in nature; rapid adaptation can be facilitated by polygenic traits, whereas polygenic traits may restrict re-use of the same genes in adaptation (genetic convergence). The rapidly evolving life histories of guppies in response to predation risk provide an opportunity to test this proposition. Guppies adapted to high- (HP) and low-predation (LP) environments in northern Trinidad evolve rapidly and convergently among natural populations. This system has been studied extensively at the phenotypic level, but little is known about the underlying genetic architecture. Here, we use an F2 QTL design to examine the genetic basis of seven (five female, two male) guppy life history phenotypes. We use RAD-sequencing data (16,539 SNPs) from 370 male and 267 female F2 individuals. We perform linkage mapping, estimates of genome-wide and per-chromosome heritability (multi-locus associations), and QTL mapping (single-locus associations). Our results are consistent with architectures of many-loci of small effect for male age and size at maturity and female interbrood period. Male trait associations are clustered on specific chromosomes, but female interbrood period exhibits a weak genome-wide signal suggesting a potentially highly polygenic component. Offspring weight and female size at maturity are also associated with a single significant QTL each. These results suggest rapid phenotypic evolution of guppies may be facilitated by polygenic trait architectures, but these may restrict gene-reuse across populations, in agreement with an absence of strong signatures of genetic convergence from recent population genomic analyses of wild HP-LP guppies.

evolutionary biology↗

bioRxiv2021

Cascading indirect genetic effects in a clonal vertebrate

Understanding how individual differences among organisms arise and how their effects propagate through groups of interacting individuals are fundamental questions in biology.Individual differences can arise from genetically-based variation in the conspecifics with which an individual interacts, and these effects might then be propagated to other individuals. Using a clonal species, the Amazon molly (Poecilia formosa), we test the hypothesis that such indirect genetic effects (IGE) propagate beyond individuals that experience them firsthand. We tested this hypothesis by exposing genetically identical Amazon mollies to social partners of different genotypes, and then moving these individuals to new social groups in which they were the only member to have experienced the IGE. We found that genetically different social environments induced different levels of aggression experienced by the focal animals, and that these genetically-based social effects carried over into new social groups to influence the behavior of individuals that did not directly experience the previous social environments. Our data reveal that IGE can cascade beyond the individuals that directly experience them to influence phenotypes even when there is no genetically-based variation present within interacting groups. Theoretical and empirical expansion of the quantitative genetic framework developed for IGE to include cascading and other types of carry-over effects will improve understanding of social behavior and its evolution.

animal behavior and cognition↗

bioRxiv2020

Colormesh: A novel method for quantifying variation in complex color patterns

Color variation is one of the most obvious examples of variation in nature. Objective quantification and interpretation of variation in color and complex patterns is challenging. Assessment of variation in color patterns is limited by the reduction of color into categorical measures and lack of spatial information. We present Colormesh as a novel method for analyzing complex color patterns that offers unique capabilities. Compared to other methods, Colormesh maintains the continuous measure of color at individual sampling points throughout the pattern. This is particularly useful for analyses of variation in color patterns, whether interest is in specific locations or the pattern as a whole. In our approach, the use of Delaunay triangulation to determine sampling location eliminates the need for color patterns to have clearly defined pattern elements, and users are not required to identify discrete color categories. This method is complementary to several other methods available for color pattern quantification, and can be usefully deployed to address a wide range of questions about color pattern variation.

evolutionary biology↗

bioRxiv2019

Non-parallel transcriptional divergence during parallel adaptation

How underlying mechanisms bias evolution toward predictable outcomes remains an area of active debate. In this study, we leveraged phenotypic plasticity and parallel adaptation across independent lineages of Trinidadian guppies (Poecilia reticulata) to assess the predictability of gene expression evolution during parallel adaptation. Trinidadian guppies have repeatedly and independently adapted to high- and low-predation environments in the wild. We combined this natural experiment with a laboratory breeding design to attribute transcriptional variation to the genetic influences of population of origin and developmental plasticity in response to rearing with or without predators. We observed substantial gene expression plasticity as well as the evolution of expression plasticity itself across populations. Genes exhibiting expression plasticity within populations were more likely to also differ in expression between populations, with the direction of population differences more likely to be opposite those of plasticity. While we found more overlap than expected by chance in genes differentially expressed between high- and low-predation populations from distinct evolutionary lineages, the majority of differentially expressed genes were not shared between lineages. Our data suggest alternative transcriptional configurations associated with shared phenotypes, highlighting a role for transcriptional flexibility in the parallel phenotypic evolution of a species known for rapid adaptation.

evolutionary biology↗

bioRxiv2018

Untangling the role of selection and drift in population divergence via transcriptional network simulations: Extended analysis of Ghalambor et al. (2015)

Introduction Introduction Literature Cited A long-standing question in evolutionary biology is whether phenotypic plasticity influences adaptive evolutionary change (e.g. Waddington 1961; Price et al. 2003; Ghalambor et al. 2007; Lande 2009; Wund 2012). The same genotype can produce different phenotypes in response to different environments, but whether such plasticity constrains or facilitates evolutionary change remains an unresolved and controversial problem (Ghalambor et al. 2007). Theoretical models to date have made diverse predictions on the role of plasticity in evolutionary divergence (e.g. Ancel 2000; Paenke et al. 2007; Thibert-Plante and Hendry 2011), and empirical studies have largely been limited to retrospective approaches that infer the role of plasticity long-after popu ...

evolutionary biology↗