Biology subjects

Harrison, L. M.

Publications and source records attributed to Harrison, L. M..

7 recordsLinked to original sources

bioRxiv2026

N-glycoproteomic profiling of the hookworm Ancylostoma ceylanicum reveals heterogeneity and phosphorylcholine-modified glycans in larval and adult parasites.

Glycans of parasitic worms (helminths) have been previously described with roles in immunomodulation and the recognition of helminth proteins by the host immune system. Despite their significance in other systems, little is known about the glycosylation patterns of the hookworm Ancylostoma ceylanicum, an intestinal parasite of animals and humans, due to challenges in sample collection and structural complexity of many nematode glycans. Herein, we describe results of a released N-glycan analysis of adult hookworms, which revealed oligomannose, hybrid, and complex structures, including those with non-mammalian features. Next, we report the results of glycoproteomic analyses to identify hookworm glycoproteins and their associated glycan structures. We compared three enrichment methods and found that strong anion-exchange electrostatic repulsion liquid chromatography (SAX-ERLIC) led to the identification of the most N-glycopeptides. Analyses of larval and adult extracts revealed 82 larval and 157 adult N-glycoproteins, the majority of which were stage-specific. Of particular interest, phosphorylcholine (PC), a known immunomodulatory glycan modification, was detected in both stages, and several unique PC-modified structures were identified in each. Collectively, these results provide the first site-specific characterization of intact N-glycopeptides in A. ceylanicum and, with an adult-stage released glycan analysis, establish a method for glycoproteomic characterizations of other helminths.

microbiology↗

bioRxiv2026

Laboratory adaptation and complete genome assembly of a Beposo, Ghana strain of the human hookworm Necator americanus

Laboratory models are invaluable tools for studying parasite biology and pathogenesis, especially for helminth infections. However, the complex life cycles and frequently narrow host specificity of helminths present challenges to maintaining access to critical parasite material in a laboratory setting. This is especially true of Necator americanus, the most common species of hookworm that infects humans globally. Here we report the successful laboratory adaptation of an African strain of N. americanus, originally isolated from infected individuals in Beposo, Ghana. The Beposo strain has been successfully passaged across 9 generations in Golden Syrian hamsters maintained on oral dexamethasone. Differential susceptibility to mebendazole and albendazole was evaluated using an egg hatch assay, and DNA sequencing of the beta-tubulin isotype 1 gene did not identify known resistance-associated mutations in the endemic strain. Sequencing of the mitochondrial COX1 gene revealed that specimens of N. americanus from Ghana, along with reported sequences from Togo, are distinct from those from South America and Asia. Complementary microsatellite-based population analysis revealed substantial genetic variation in the founding parasite population. To further characterize the novel Beposo strain, a draft hybrid genome assembly was generated from genomic DNA extracted from a single adult male worm via an optimized Oxford Nanopore Technologies MinION library preparation approach tailored to low-input sample types. This high-quality assembly, including a complete mitogenome, is 202.8Mb in 950 contigs with an N50 >449 kb. It contains >95% of conserved nematode orthologs in complete single copy and is estimated by homology-based gene prediction to contain 12,804 genes. This study represents the first comprehensive characterization of a strain of N. americanus originating in Africa that has been successfully adapted to a laboratory animal model.

microbiology↗

bioRxiv2024

Glucocorticoids desensitize CRH neurons to norepinephrine via rapid nitrosylation-dependent regulation of α1 adrenoreceptor trafficking

Noradrenergic afferents to hypothalamic corticotropin releasing hormone (CRH) neurons provide a major excitatory drive for somatic stress activation of the hypothalamic-pituitary-adrenal (HPA) axis. We showed that glucocorticoids rapidly desensitize CRH neurons to norepinephrine and suppress inflammation-induced HPA activation via a glucocorticoid receptor- and endocytosis-dependent mechanism. Here, we show that 1 adrenoreceptor (AR1) trafficking is regulated by convergent glucocorticoid and nitric oxide synthase signaling mechanisms. Live-cell imaging of AR1b-eGFP-expressing hypothalamic cells revealed rapid corticosterone-stimulated redistribution of internalized AR1 from rapid recycling endosomes to late endosomes and lysosomes via a nitrosylation-regulated mechanism. Proximity assay demonstrated interaction of glucocorticoid receptors with AR1b and {beta}-arrestin, and showed corticosterone blockade of norepinephrine-stimulated AR1b/{beta}-arrestin interaction, which may prevent AR1b from entering the rapid recycling endosomal pathway. These findings demonstrate a rapid glucocorticoid regulation of G protein-coupled receptor trafficking and provide a molecular mechanism for rapid glucocorticoid desensitization of noradrenergic signaling in CRH neurons.

neuroscience↗

bioRxiv2024

Modeling Diffusive Search by Non-Adaptive Sperm: Empirical and Computational Insights

During fertilization, mammalian sperm undergo a winnowing selection process that reduces the candidate pool of potential fertilizers from [~]106-1011 cells to 101-102 cells (depending on the species). Classical sperm competition theory addresses the positive or stabilizing selection acting on sperm phenotypes within populations of organisms but does not strictly address the developmental consequences of sperm traits among individual organisms that are under purifying selection during fertilization. It is the latter that is of utmost concern for improving assisted reproductive technologies (ART) because low fitness sperm may be inadvertently used for fertilization during interventions that rely heavily on artificial sperm selection, such as intracytoplasmic sperm injection (ICSI). Importantly, some form of sperm selection is used in nearly all forms of ART (e.g., differential centrifugation, swim-up, or hyaluronan binding assays, etc.). To date, there is no unifying quantitative framework (i.e., theory of sperm selection) that synthesizes causal mechanisms of selection with observed natural variation in individual sperm traits. In this report, we reframe the physiological function of sperm as a collective diffusive search process and develop multi-scale computational models to explore the causal dynamics that constrain sperm fitness during fertilization. Several experimentally useful concepts are developed, including a probabilistic measure of sperm fitness as well as an information theoretic measure of the magnitude of sperm selection, each of which are assessed under systematic increases in microenvironmental selective pressure acting on sperm motility patterns. Author SummaryFertilization in mammals requires many orders of magnitude more sperm than eggs to be fertilized. This study models collective sperm movement as a diffusive search process, combining experiments and simulations to explore how heterogeneity in motility patterns and microenvironmental complexity shape fertilization success. We introduce simple metrics to quantify sperm fitness and the magnitude of selection pressure imposed by the microenvironment, revealing that sperm phenotype distributions interact with environmental constraints to determine the range of sperm traits that ultimately support successful egg contact. These insights improve our understanding of sperm population dynamics and offer practical tools for optimizing assisted reproductive technologies in clinical and agricultural settings.

cell biology↗

bioRxiv2024

A Low-Cost Microfluidics System for Light Microscopy Experiments

Microfluidics devices are powerful tools for studying dynamic processes in live cells, especially when used in conjunction with light microscopy. There are many applications of microfluidics devices including recording dynamic cellular responses to small molecules or other chemical conditions in perfused media, monitoring cell migration in constrained spaces, or collecting media perfusate for the study of secreted compounds in response to experimental inputs/manipulations. Here we describe a configurable low-cost (channel-based) microfluidics platform for live-cell microscopy, intended to be useful for experiments that require more precision/flexibility than simple rubber spacers, but less precision than molded elastomer-based platforms. The materials are widely commercially available, low-cost, and device assembly takes only minutes.

bioengineering↗

bioRxiv2024

Fluctuating salinity during development impacts fish productivity

Climate change is elevating salinity levels in many freshwater systems, and more erratic rainfall is increasing variation in salinity. Consequently, many species now experience more extreme developmental environments. Resultant shifts in developmental trajectories could change key life history traits that persist into adulthood. To date, however, how variation in salinity affects the life histories of freshwater species has been neglected despite its implications for fisheries. We ran a large-scale experiment with a global pest, the mosquitofish (Gambusia holbrooki), and manipulated the salinity experienced by juveniles: freshwater (0{per thousand}), stable salinity (10{per thousand}) or fluctuating salinity (0-20{per thousand}; mean = 10 {per thousand}). Fish developing in stable, high salinity grew faster and matured earlier, albeit with a decline in male telomeres and female gut development. Stable high salinity resulted in larger adult body size in females, but not males, which increased female fecundity. Conversely, fluctuations in salinity induced fish to grow more slowly and lowered female fecundity. Crucially, several of the long-term effects of salinity fluctuations were sex-specific, more adversely affecting females than males. We highlight that environmental variability alters an organisms vulnerability to stressors, with implications that should be considered if we wish to understand the impact of climate change on population dynamics.

ecology↗

bioRxiv2020

Acute Stress Desensitizes Hypothalamic CRH Neurons to Norepinephrine and Physiological Stress

Noradrenergic afferents to corticotropin releasing hormone (CRH) neurons of the hypothalamic paraventricular nucleus (PVN) provide a major excitatory drive to the hypothalamic-pituitary-adrenal (HPA) axis via 1 adrenoreceptor activation. The ascending noradrenergic afferents are recruited preferentially by physiological, rather than psychological, stress modalities. Glucocorticoids secreted in response to HPA activation feed back onto the hypothalamus to negatively regulate the HPA axis, providing a critical autoregulatory constraint that prevents glucocorticoid overexposure. Whether differential negative feedback mechanisms target stress modality-specific HPA activation is not known. Here, we reveal a desensitization of the 1 adrenoreceptor activation of the HPA axis following acute stress that is mediated by rapid glucocorticoid regulation of adrenoreceptor trafficking. Prior stress desensitized the HPA axis to subsequent physiological, but not psychological, stress. Our findings demonstrate rapid glucocorticoid suppression of adrenoreceptor signaling in CRH neurons that is specific to physiological stress activation, and reveal, therefore, a rapid, modality-selective glucocorticoid feedback mechanism.

neuroscience↗