Real-time Characterisation and modulation of microbe-induced inflammation and resolution, using a novel zebrafish larval corneal injury and infection model

Real-time Characterisation and modulation of microbe-induced inflammation and resolution, using a novel zebrafish larval corneal injury and infection model | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Real-time Characterisation and modulation of microbe-induced inflammation and resolution, using a novel zebrafish larval corneal injury and infection model Kelvin Cheng, Carl Tucker, Justyna Cholewa-Waclaw, Stephen Mitchell, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7524577/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Microbial keratitis (MK) is a major global cause of blindness. Yet, treatment is still heavily dependent on antimicrobials with limited options for immunomodulators - despite the critical role of dysregulated immune responses in disease pathogenesis. This gap reflects a critical unmet clinical need and is compounded by the lack of model systems capable of real-time high-resolution immune dynamics analysis. To address this, we developed a zebrafish larvae MK model utilising transgenic zebrafish lines with fluorescently labelled neutrophils, macrophages and basal epithelial cells. Corneal injury triggered rapid immune cell recruitment which was amplified by exposure to pro-inflammatory mediators such as N-formylmethionine-leucyl-phenylalanine (fMLF) and leukotriene B4 (LTB4). Infection with live bacteria induced robust, sustained neutrophil and macrophage recruitment, marked by increased neutrophil speed and migratory distance. This model enables dynamic in-vivo visualization of immune cell dynamics, offering a powerful and scalable platform to accelerate the discovery and screening of novel immunomodulators for MK. Health sciences/Medical research/Experimental models of disease Biological sciences/Immunology/Infection Health sciences/Medical research/Preclinical research Biological sciences/Immunology/Innate immunity Biological sciences/Immunology/Inflammation Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SuppFigure1.pdf Scanning electron microscopy images of injured zebrafish larvae corneas. SuppFigure2.pdf Validation of zebrafish larvae corneal injury model. SuppFigure3.pdf Indices of motility of neutrophils and macrophages in sterile injury with or without infection with P. aeruginosa. SuppVideo1.avi Widefield timelapse of a Tg(K19:GFP; LysC:mTurquoise) zebrafish larvae post-injury demonstrating rapid wound closure followed by accumulation of neutrophils within the cornea. SuppVideo2.avi Maximum intensity projection timelapse of a Tg(K19:GFP, LysC:mTurquoise; mfap4:tdTomato-CAAX) zebrafish larvae post-injury demonstrating rapid wound closure, followed by accumulation of neutrophils an SuppVideo3.mp4 Representative timelapse of injured corneas at 1 h-post injury with or without infection. SuppVideo4.mp4 Three-dimensional reconstruction of injured and cornea of 3 day-post-fertilisation larvae infected with P. aeruginosa at 1 h post-infection demonstrating distribution of P. aeruginosa in relation to i Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7524577","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":536171733,"identity":"972ba90f-8c25-445e-a7a5-6e868b53c691","order_by":0,"name":"Kelvin 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