Biology subjects

Qu, J.

Publications and source records attributed to Qu, J..

At least 19 recordsLinked to original sources

bioRxiv2026

Yck2 links mitochondrial function and pH homeostasis to cell wall remodeling and antifungal susceptibility in Cryptococcus neoformans

Invasive fungal infections are on the rise due to climate change, antifungal resistance, and increased usage of immunomodulating therapies. The antifungal arsenal is limited in number and in efficacy necessitating novel therapeutics. Deletion of the gene encoding the fungal yeast casein kinase, YCK2, has pleiotropic effects, impacting morphology, drug resistance, metabolism, cell wall, and virulence. Yck2 has recently been shown to be druggable, however the mechanisms by which Yck2 elicits its impacts are unknown. We show that C. neoformans Yck2 is a regulator of cell wall masking, thermotolerance, general stress, and drug resistance. Proximity labeling of Yck2-interacting proteins revealed that Yck2 interacts with proteins that function in the mitochondria as well as the essential plasma membrane H+ ATPase, Pma1. A role for Yck2 in mitochondrial regulation was supported by sensitivity to mitochondrial inhibitors and increased mitochondrial ROS production in the absence of Yck2, and presence of Yck2 in mitochondrial fractions. Using pHluorin-expressing cells we also found that intracellular pH is elevated in cells lacking Yck2 supporting a role in regulating Pma1 activity. Both mitochondria and cellular pH may contribute to cellular signaling and may explain the multitude of phenotypes associated with the yck2{Delta} mutant. Our results begin to identify the mechanisms by which Yck2 contributes to fungal cellular homeostasis which may support efforts to optimize Yck2 inhibitors.

microbiology↗

bioRxiv2026

TEAD1 signaling modulates adrenal chromaffin maturation

Chromaffin cells synthesize and secrete catecholamines to coordinate systemic stress responses and regulate diverse neuroendocrine and metabolic functions. However, the molecular mechanisms governing chromaffin-cell differentiation and their disruption in pheochromocytoma (PC) remain incompletely understood. Here, through integrated analyses of human developmental atlases, patient-derived transcriptomic datasets, genetically engineered mouse models, and chromaffin organoids, we identify TEAD1 signaling as a critical regulator of chromaffin-cell differentiation and function. In vivo studies using a chromaffin cell-specific TEAD1 overexpression mouse model demonstrated that suppression of TEAD signaling markedly compromises chromaffin-cell differentiation and endocrine function. Additionally, compared with other TEAD family members, TEAD1 transcriptional activities are readily affected by sequences near the binding motif. To identify therapeutically actionable regulators of TEAD1 signaling, we established a TEAD activity-based screening platform and identified the serotonin receptor HTR5A antagonist SB699551 as a potent modulator of chromaffin-cell state. SB699551 suppressed PC-cell proliferation in vivo, and remodeled catecholamines synthesis in primary human PC cells. Additionally, application of SB699551 to human PC tumor revealed a subpopulation of primary chromaffin cells sensitive to this compound. Mechanistically, CXXC5 and L1CAM were identified as downstream SB699551-TEAD1 signaling effectors mediating chromaffin-cell proliferation and differentiation. Overall, we demonstrate that TEAD1 signaling is a fundamental mechanism regulating chromaffin cell differentiation and that modulation of TEAD1 signaling via SB699551 offers a new area of investigation in chromaffin cell biology.

molecular biology↗

bioRxiv2026

MagicLamp: a web server and software toolkit for targeted gene annotation of microbial functions

Genome and metagenome annotation tools designed for large databases are ill-suited to the discovery of specialized, ecologically relevant microbial functions. MagicLamp (https://github.com/Arkadiy-Garber/MagicLamp) is a modular command-line software toolkit that performs targeted functional gene annotation searches using curated collections of hidden Markov models (HMMs), each representing discrete microbial metabolic processes. MagicLamp is also available as a web server: https://midauthorbio.com/#magiclamp. This targeted approach enables sensitive and specific annotation of genes involved in defined microbial processes, allowing MagicLamp to serve as a dedicated repository for the annotation of specialized microbial functions currently overlooked in other databases and software. The server accepts unannotated genome assemblies or GenBank-formatted annotations to perform HMM-based searches against curated model sets with reproducible, model-specific bit-score thresholds. Automated results are returned as tabular summaries and interactive HTML reports containing cross-genome/metagenome comparisons.

bioinformatics↗

bioRxiv2026

Quantifying annotation-stratified pleiotropy and co-polygenicity between complex traits

Understanding shared genetic architecture is essential to interpreting disease comorbidities and trait correlations. We introduce SBayesAPP, a Bayesian model that integrates GWAS summary statistics with functional annotations to jointly estimate annotation-stratified SNP effect-size correlation and pleiotropic variant proportion (co-polygenicity) between traits, dissecting genetic correlation and coheritability enrichment across annotations. Simulations and real data analyses show improved accuracy and interpretability over existing methods. In type 2 diabetes analyses with 15 traits, SBayesAPP reveals clear tissue- and cell-type-specific enrichment and distinguishes mechanisms driven by few large-effect variants versus many modest-effect variants. The analysis of smoking and lung cancer prioritizes lung and immune cells, and identifies cell-type-specific genetic correlations driven by either pleiotropic or lung-cancer-specific variants, consistent with a causal relationship model. For schizophrenia and educational attainment, despite near-zero genome-wide genetic correlation, cell-type-specific correlations range from -0.20 to 0.21, with strong (co)heritability enrichment and high co-polygenicity found in dopaminergic neurons and oligodendrocytes. These results highlight the ability of SBayesAPP to resolve annotation-specific genetic sharing and uncover biological mechanisms across complex traits.

genomics↗

bioRxiv2026

L Fucose Dependent Biofilm Formation by Escherichia coli Enhances Polymicrobial Interactions and Antibiotic Tolerance on Urinary Catheters

Urinary tract infections are common healthcare associated infections, a large subset of which are caused by indwelling catheters. Long term catheterization causes persistent, asymptomatic, polymicrobial colonization despite catheters changes and antibiotic usage. In these polymicrobial populations, P. mirabilis, E. faecalis, and E. coli were found as the most common co-colonizing species. We investigated how interactions between P. mirabilis, E. coli, and E. faecalis contribute to biofilm formation and colonization of urinary catheters. Our results show that the interaction between these three species leads to enhanced biofilm biomass driven by an increase in total protein content of the biofilm. Biofilm enhancement required all three species and was also media-dependent, especially for dual-species combinations. Importantly, triple species biofilms also demonstrate biofilm enhancement when established under flow conditions in a biofilm reactor model using silicone urinary catheters. Additionally, triple species biofilm enhancement occurred in co-colonizing isolates from catheterized patients and was found to be specific to interactions between these three species. Triple species biofilms also demonstrated a species-dependent resistance to two commonly used antibiotics, ciprofloxacin and nitrofurantoin. By examining priority effects, E. coli was found to be the main facilitator of biofilm enhancement in a flow model. Finally, proteomics revealed that an L-fucose utilization pathway in E. coli was a key contributor to triple species biofilm enhancement. Overall, our results demonstrate the significant impact of polymicrobial interactions on biofilm formation in the catheterized environment and highlight ways in which complex microbial interplay and priority effects can shape the establishment of persistent colonization.

microbiology↗

bioRxiv2026

BTEXgenie: A curated and user-friendly tool for profile HMM-based substrate-specific annotation of BTEX degradation genes

BackgroundBenzene, toluene, ethylbenzene, and xylene (BTEX) are volatile aromatic hydrocarbons that are widespread environmental pollutants arising from petroleum processing, fuel combustion, and other industrial activities. Persistent BTEX contamination poses substantial risks to human health and ecosystems, underscoring the need for effective long term remediation strategies. Microbial bioremediation is a promising and sustainable approach for BTEX removal, but development of these approaches requires accurate detection of the genes and pathways responsible for substrate specific degradation. Although profile hidden Markov model (HMM) databases are widely used for functional annotation, existing annotation resources lack the substrate-specific resolution needed to distinguish between closely-related BTEX-degrading enzymes with different catalytic specificities. ResultsWe developed BTEXgenie as a sensitive annotation tool that uses custom HMMs built from alignments of experimentally validated BTEX degradation proteins to identify genes involved in the initial steps of aerobic and anaerobic BTEX degradation. BTEXgenie improved detection of anaerobic BTEX degradation genes that were absent from KOfam annotations. In benchmarking against the KEGG KOfam HMM database, BTEXgenie achieved 17.73%higher overall sensitivity while maintaining comparable specificity at 97.02%across genes involved in BTEX degradation pathways. When applied to environmental metagenomes, BTEXgenie recovered pathway patterns consistent with reported site characteristics and known degradation potential. In addition to gene annotation, BTEXgenie supports downstream interpretation through KEGG pathway-based visualization of detected functions and Circos-based visualization of genomic hit distributions. ConclusionsBTEXgenie is a substrate-specific annotation tool built from custom HMMs for detecting genes involved in BTEX degradation. By integrating gene annotation with pathway and genome-level visualizations, BTEXgenie facilitates characterization of microbial BTEX degradation potential in environmental and comparative genomic studies.

bioinformatics↗

bioRxiv2026

Hex-MASP for Mapping the Whole-tissue Spatial Proteome and the Intra-brain Distribution of Monoclonal Antibodies

Whole-tissue spatial proteomics level provides critical insights into region-specific biological regulations but remains challenging. Previously, we introduced the Micro-scaffold Assisted Spatial Proteomics (MASP) concept for whole-tissue mapping. However, this prototype required substantial development in spatial resolution, practicality, and throughput for practical application. Here we present a next-generation MASP technique (hex-MASP) featuring i) a new design of hexagonal-micro-wells fabricated with optimized Projection Micro-Stereolithography (P{micro}SL) 3D-printing, achieving high spatial resolution, sampling robustness and mechanical strength for reproducibly compartmentalizing even tough tissues; ii) enhanced throughput/effectiveness in sample preparation and LC-MS analysis with high quantitative quality. Applied to mouse brain, hex-MASP for the first time achieved in-depth, whole-tissue mapping for >6,000 proteins in mouse brains, with high spatial accuracy and excellent data quality. The substantially improved resolution revealed critical regional details across the entire brain, that were not previously captured, enabling precise depiction of protein distribution heterogeneity. This technique enabled the discovery of many unreported regionally-enriched proteins across brain structures. We further applied hex-MASP to investigate the intra-brain distribution of intracerebroventricularly-dosed antibody therapeutics and related proteins, which to our knowledge, enabled whole tissue mapping of protein drugs for the first time and revealed novel mechanistic insights into antibody distribution and localized treatment effects. Hex-MASP represent a robust, scalable platform for whole-tissue spatial proteomics.

pharmacology and toxicology↗

bioRxiv2026

Optical Windows for Transcranial Brain Imaging in Living Mice: Skull Thinning, Clearing, and Beyond

Longitudinal, noninvasive in vivo imaging is essential for studying brain physiology and pathological mechanisms. Advances in transcranial optical windows, including thinned-skull and optical clearing techniques, have markedly improved imaging depth and resolution when combined with multiphoton microscopy. However, their optical performance often deteriorates rapidly, and quantitative studies on long-term stability and the causes of image quality loss remain limited. In this work, we systematically investigated current transcranial window approaches using multiphoton excited fluorescence microscopy (MPEFM) and adaptive optics, examining longevity, optical aberrations, and imaging resolution. Our results reveal that progressive skull regrowth is a fundamental limitation across all window types, leading to substantial declines in signal quality and resolution for conventional MPEFM thereby reducing achievable high-resolution imaging depth. To address these challenges, we developed a localized glucocorticoid (GC) delivery strategy that significantly extends window performance for up to one month. Furthermore, we demonstrated that a GC-loaded hydrogel sealing method effectively suppressed skull regrowth while preserving optimal optical properties, offering a potential and practical route to chronic, high-fidelity transcranial imaging. These findings provide mechanistic insight into window degradation and establish a framework for sustained, long-term in vivo brain imaging.

neuroscience↗

bioRxiv2026

p63 and p73 regulate convergent and factor-specific transcriptional programs in cutaneous squamous cell carcinoma

Aberrant transcriptional regulation is a defining feature of squamous cell carcinoma (SCC), yet how lineage transcription factors coordinate shared and factor-specific oncogenic programs remains poorly understood. Although TP63 (p63) is frequently amplified in SCC, the contribution of its paralog TP73 (p73) has remained unclear. Here we show that p73, together with p63, is upregulated in skin SCC and is required for tumorigenesis. Mechanistically, p63 and p73 form heteromeric complexes and co-occupy distal enhancer elements, establishing a shared chromatin regulatory framework. Integration of chromatin and transcriptomic profiling reveals that this common enhancer landscape supports both convergent and divergent transcriptional outputs. Both factors cooperatively sustain core proliferation but also exert regulatory biases, with p63 preferentially reinforcing epithelial lineage circuits and p73 contributing to DNA replication and stress-associated pathways. Among shared downstream targets, p63/p73 co-regulation of multiple epidermal growth factor receptor (EGFR) ligands establishes a feed-forward signaling module that amplifies mitogenic signaling. Amphiregulin emerges as a dominant functional mediator, and its depletion phenocopies key aspects of p63/p73 loss, including impaired proliferation and tumor formation. Together, these findings support a model in which shared enhancer occupancy by p63 and p73 drives cooperative and factor-specific transcriptional programs, linking chromatin regulation to signaling and tumor maintenance.

molecular biology↗

bioRxiv2026

Functional activation reveals a repressor-gated fourth type VI secretion system in clinical Pseudomonas aeruginosa

Pseudomonas aeruginosa is a major opportunistic pathogen in which the type VI secretion system (T6SS) contributes to interbacterial competition and virulence. While most strains encode three T6SSs, we here functionally characterize a fourth system (H4-T6SS) in a clinical P. aeruginosa isolate. The H4-T6SS gene cluster is transcriptionally silent under laboratory conditions. By promoter rewiring, we demonstrate that the H4-T6SS assembles a contractile machine and mediates effector secretion and robust inter- and intraspecific antagonism. We identify a dominant pore-forming effector and its secretion pathway and a silent yet conserved nuclease effector. Genetic and biochemical analyses show that H4-T6SS expression is activated by an H4-encoded transcriptional regulator and repressed by H-NS family proteins. Analysis of over 1,200 clinical isolates reveals that the H4-T6SS is distributed across lineages and geographic regions. Finally, we observe functional crosstalk between H4- and H2-T6SSs. Together, these findings establish H4-T6SS as a functional and repressor-gated secretion system that expands the competitive arsenal of clinical P. aeruginosa.

microbiology↗

bioRxiv2026

A Physics-based Model Reveals Mechanisms of Epigenetic Memory

In multi-cellular eukaryotic organisms, cell type and specific functional identity are defined by the epigenetic patterning of chemical modifications to DNA and chromatin that modulate the expression and silencing of specific genes. When a cell divides, histones containing important epigenetic marks are distributed between the two daughter strands leading to a temporary dilution of epigenetic information and cell identity. In this work we introduce a physics-based model of epigenetic memory that explains how cells restore and maintain H3K9me3 and H3K27me3 histone methylation patterning after cell division. We demonstrate that emergence and maintenance of the epigenetic program is driven by an evolved mechanism that makes use of the biophysics of polymers, phase condensates and enzymatic activity. We validate our model via genome-wide epigenetic time-course simulation and comparison to experimental epigenetic data from multiple donors, multiple cell types, and for multiple epigenetic marks. Finally, we use our model as a conceptual framework to understand cellular reprogramming by hypothesizing that these processes first contend with and later utilize somatic epigenetic maintenance programs.

biophysics↗

bioRxiv2026

Complement Factor H (Y402H) polymorphism for age-related macular degeneration alters retinal lipids

Age-related macular degeneration (AMD), the leading cause of blindness in the elderly, is associated with multiple risk factors and involves death of the retinal pigment epithelium (RPE). We investigated how the Y402H polymorphism of Complement Factor H (CFH) and cigarette smoke extract (CSE), major AMD genetic and environmental risks, affect lipid metabolism in RPE differentiated from induced pluripotent stem cells (iPSC-RPE) that were derived from human donors genotyped for low-risk (LR) or high-risk (HR) CFH. Results from discovery-based (lipidomics, proteomics) and targeted (mitochondrial fatty acid oxidation (FAO)) assays found significant genotype-dependent differences under basal conditions include higher free fatty acids and cholesterol esters in HR cells. CSE induced differences in proteins regulating lipid handling, lipolysis, and inflammation. Lower FAO in HR cells was observed in multiple donors and pairs of parent/isogenic edited lines compared with LR lines. CSE induced lipid accumulation, lipid composition remodeling, and upregulation of proteins involved in lipid synthesis/hydrolysis, production of bioactive lipid mediators, and metabolism of ceramide and cholesterol. These results elucidate putative mechanisms driving pathology in RPE harboring CFH Y402H.

systems biology↗

bioRxiv2025

Integrating Longitudinal Metabolite Profiles Improves Trait Prediction in Pigs in a Trait- and Timepoint-Dependent Manner

BackgroundAccurate prediction of genetic merit is essential for accelerating genetic improvement in pigs, particularly for traits that are costly or difficult to measure directly. This study investigated the potential of integrating individual-level blood serum metabolite profiles sampled at two developmental stages (10-week and 20-week) into genomic prediction models for five economically important traits: average daily feed intake (DFI), feed conversion ratio (FCR), backfat thickness (BF), test daily gain (TDG), and loin depth (LD). Using a BayesC modeling framework, we analyzed 1,637 pigs from a single purebred population with complete phenotype, genotype, and metabolite-profile data. We evaluated seven models, including a genotype-only model (G), metabolite-only models using metabolite-profile data from 10-week (M1), 20-week (M2), or both timepoints (M1+M2), and combined models integrating genotypes with metabolite-profile data (G+M1, G+M2, G+M1+M2). ResultsHeritability estimates were generally low to moderate, ranging from 0.07 to 0.30 for the 10-week metabolite profile and from 0.04 to 0.28 for the 20-week metabolite profile. Prediction accuracy of phenotype consistently improved when metabolite-profile data were integrated into genotype-based models, although the magnitude of improvement varied depending on the trait and the timepoint of metabolite sampling. Prediction accuracy increased from 0.31 (G) to 0.41 (G+M2; G+M1+M2) for DFI, and from 0.27 (G) to 0.33 (G+M2; G+M1+M2) for FCR. The latter two models also delivered the largest gains over G for BF (from 0.41 to 0.45) and TDG (from 0.28 to 0.32). However, LD benefited the most when both 10-week and 20-week metabolite profiles were combined (G+M1+M2: 0.45 compared to G: 0.42). ConclusionsAcross all traits, models combining genotype data with metabolite profiles from one or multiple timepoints achieved the highest or equally high prediction accuracies compared to the genotype-only model, reflecting complementary biological insights captured by metabolite profiles. These findings highlight the potential value of metabolite-profile data as an intermediate omics layer to enhance genomic prediction, particularly when integration strategies are tailored to trait-specific biology and sampling timepoints.

genetics↗

bioRxiv2025

Universal super-resolution for subcellular fluorescence imaging

Super-resolution fluorescence microscopy has emerged as an essential instrument for investigating subcellular structures and monitoring intracellular physiology. Despite rapid advances driven by deep learning, most existing approaches are modality specific and exhibit limited generalizability, restricting their practical applicability. Here we introduce universal super-resolution (UniSR), a framework that operates across microscopes and subcellular structures without optical modifications or prior sample knowledge. UniSR is initially pre-trained on simulated image pairs to establish mappings from low-to high-resolution features and subsequently fine-tuned with only one experimental image pair, reducing data demand and acquisition time. Across wide-field and point-scanning modalities, UniSR enhances resolution to match structured illumination, single-molecule localization, or stimulated emission depletion microscopy. Applied to diverse biological samples, it reveals nanoscale structures, dynamic processes, and organelle interactions with high spatiotemporal fidelity. UniSR provides a practical and broadly generalizable tool for super-resolution imaging in live-cell research.

bioinformatics↗

bioRxiv2025

PET-Microplastics Trigger Endothelial Glycocalyx Loss via ER Stress and ROS Unleashing IL-1β-Driven SMC Switching and Early Aortic Structural Impairment

Polyethylene terephthalate microplastics (PET-MPs), a major microplastics component identified in human vasculature, pose emerging environmental health risks. This study systemically profiled MPs in human aortic tissues and investigated the mechanisms underlying PET-MPs-induced aortic injury in vivo and in vitro. Chronic oral exposure of Sprague-Dawley rats to PET-MPs (1.0-100 mg/L) resulted in endothelial glycocalyx loss and structural impairment of aortic elastic fibers, with MPs accumulating within aortic endothelial cells. Transcriptomic and biochemical analyses revealed that PET-MPs triggered endoplasmic reticulum stress (ERS) and reactive oxygen species (ROS) generation in human aortic endothelial cells (HAECs), driving glycocalyx degradation and NF-{kappa}B-mediated inflammation. Proteomic profiling identified endothelial-derived IL-1{beta} as a key mediator, which subsequently induced phenotypic switching in human aortic smooth muscle cells (HASMCs) in vitro. Pharmacological inhibition of ERS (TUDCA), ROS (NAC), or IL-1{beta} (Canakinumab) attenuated this pathogenic cascade. Crucially, restoration of the glycocalyx using Sulodexide mitigated endothelial dysfunction and downstream HASMC phenotypic switching. These findings establish endothelial glycocalyx degradation via ERS-ROS as a novel mechanism for PET-MPs-induced vascular injury and highlight glycocalyx protection as a potential strategy against environmental microplastic hazards.

pharmacology and toxicology↗

bioRxiv2025

HIF-1α and RhoA Drive Enhanced Motility and Aerotaxis of Polyaneuploid Prostate Cancer Cells in Hypoxia

Most cancer deaths result from metastasis, yet only a rare subset of tumor cells can complete this process. Among these, polyaneuploid cancer cells (PACCs), which arise via endoreplication under stressors such as hypoxia, are implicated as metastatic drivers, but how they acquire this potential is poorly understood. Here, we show that prostate cancer-derived PACCs exhibit features predictive of invasion and intravasation. Time-lapse fluorescence microscopy and single-cell tracking under hypoxia revealed that PACCs migrated significantly farther than Non-PACCs, consistent with local invasion. PACC trajectories showed a strong tendency to migrate toward oxygen, consistent with aerotaxis and predictive of intravasation. siRNA-mediated knockdown demonstrated that the enhanced motility and aerotaxis of hypoxic PACCs require both HIF-1and RhoA, with RhoA expression suppressed upon HIF-1 inhibition. Altogether, we propose a HIF-1[->] RhoA [->] motility/aerotaxis mechanism enabling PACCs to escape hypoxic cores, invade tissue, and access vasculature, highlighting them as a uniquely invasive subpopulation with implications for anti-metastatic therapies.

cancer biology↗

bioRxiv2025

Ceramide-induced Endoplasmic Reticulum Stress as a Targetable Vulnerability in Endocrine Therapy-Resistant Breast Cancer

Despite the success of endocrine therapy (ET) in treating hormone receptor-positive breast cancer, a significant proportion of patients relapse during or after treatment, making ET resistance a major clinical challenge. Previously we have shown that ET-resistant breast cancer cells exhibit reduced ceramide levels and an increased sensitivity to ceramide-induced cell death. Here, we demonstrate that ceramides induce a distinct transcriptional reprogramming in ET-resistant cells, characterized by upregulation of endoplasmic reticulum stress (EnRS) pathways. Ceramide-induced EnRS is PERK-dependent and functionally linked to cell death in multiple models of ET resistance. Using a photoactivatable ceramide probe, we identify TRAM1 as a functionally important ceramide-interacting protein (CIP) in ET-resistant cells that correlates with worse relapse-free survival and a more aggressive breast cancer phenotype in luminal breast cancer patients. Additionally, knockdown of TRAM1 phenocopies ceramide action in ET resistance, thereby suggesting its role in mediating ceramide-induced lethal actions in ET resistance. Together, our findings reveal that ET-resistant breast cancer cells are more sensitive to PERK-mediated EnRS as compared to their ET-sensitive counterparts. Ceramides can exploit this dependence by interacting with CIPs such as TRAM1, leading to PERK activation and consequential cell death preferentially in the ET-resistant breast cancer models.

cancer biology↗

bioRxiv2025

Dynamic Retinal Pathology in Glaucoma Progression Revealed by High-Resolution Functional Imaging in Vivo

Glaucoma, a complex and multifactorial neurodegenerative disease, is commonly associated with elevated intraocular pressure and primarily characterized by the progressive loss of retinal ganglion cells (RGCs) and their axons. Despite its prevalence, our understanding and treatment of this disease remain challenging due to the intricate interplay of various pathophysiological factors and the limited capability for in vivo functional study of the disease. In this work, we investigated the dynamic retinal pathology of glaucoma from onset to late stages through longitudinal in vivo high-resolution imaging in a silicone oil-induced ocular hypertension glaucoma mouse model. We developed an optimized adaptive optics two-photon excitation fluorescence microscopy (AO-TPEFM) technique for both morphological and functional assessments of pathological changes in the retina across three distinct glaucoma phenotypes with varying progression rates. Our AO-TPEFM technology visualized the complete process of functional and structural changes in the three most important retinal components related to glaucoma: microvascular vessels, microglia, and RGCs during disease progression. Notably, our functional imaging revealed pathological alterations in microvascular circulation, microglia state, and RGC functionalities at a very early stage of disease development when retinal morphology still appeared normal, providing critical insights into glaucoma pathogenesis. This research also demonstrates that AO-TPEFM is a powerful tool for the in vivo study of general retinal diseases.

bioengineering↗