Biology subjects

Cheng, K.

Publications and source records attributed to Cheng, K..

At least 19 recordsLinked to original sources

bioRxiv2026

Live imaging reveals oocyte membrane pre-remodeling forming permissive space for asymmetric division

Abstract Successful oocyte maturation requires first polar body (PB1) extrusion, yet how the oocyte prepares for this asymmetric event remains unclear. Live membrane imaging of mouse oocytes reveals pronounced regional heterogeneity in plasma-membrane dynamics, generating an early polarized pattern across the surface. Whereas most of the oocyte surface and its microvilli remain relatively stable, a discrete membrane domain collapses and retracts around germinal vesicle breakdown (GVBD), separating from the zona pellucida to establish a pre-extrusion space (PES). Transient oocyte comb-like protrusions arise within the PES and subsequently collapse, converting the protrusion-filled region into an open cavity before PB1 extrusion. Mechanistically, early ERK28 dependent dissociation of the cortical scaffold ZO-1 is required for this space to form. ERK2 phosphorylates ZO-1, suppresses its phase separation, and promotes its removal from the cortex. Blocking MEK-ERK signaling prevents ZO-1 dissociation, abolishes pre-extrusion space formation, and impairs PB1 extrusion, whereas ERK activation restores these events in aged oocytes. Together, our findings identify a membrane collapse-based pre-remodeling mechanism that licenses PB1 extrusion and reveal ERK-ZO-1 signaling as a key regulator of oocyte maturation.

cell biology↗

bioRxiv2026

Dissecting functional dynamics of fecal-derived in vitro stable microbial communities with metaproteomics

Fecal-derived in vitro microbial communities represent promising models for investigating gut microbiome function and facilitating translational applications. However, the factors governing community establishment and the underlying mechanisms remain incompletely understood. We employed large-scale quantitative metaproteomics to investigate the functional and community dynamics of human fecal microbiomes cultured in three media over 25 serial passages. Our results demonstrated that both culture medium and donor-specific microbiome characteristics significantly influence community establishment. Taxon-specific functional analyses revealed nutrient availability in the medium and the microbial stress responses as key determinants of in vitro community establishment. Notably, the metabolism of mucin glycans in RapidAIM medium contributed to the maintenance of Bacteroidaceae species that were poorly supported in other media, resulting in an in vitro community that most closely resembled the uncultured fecal inoculum. Overall, these findings provide mechanistic insights into in vitro gut microbial community establishment, supporting further optimization and application of fecal-derived in vitro microbiome models.

microbiology↗

bioRxiv2026

MetaPilot: genome-aware adaptive search-space refinement for unified DDA and DIA metaproteomics

Metaproteomics directly measures microbial protein expression in complex communities, but analysis is constrained by large, poorly structured search spaces. Microbial gene catalogues provide broad coverage, but their pooled organization hinders genome-level evidence accumulation and consistent DDA and DIA analysis. Here, we present MetaPilot, a genome-aware workflow that uses conserved marker-protein evidence to guide adaptive genome-resolved search-space refinement. MetaPilot maps identifications to candidate genomes, ranks them by marginal peptide contribution, and constructs refined sample-specific search spaces for final identification, quantification and multi-layer reporting. Across DDA and DIA datasets from defined mixtures and faecal microbiomes, MetaPilot adapted genome selection to sample complexity, preserved substantial overlap with published peptide identifications and expanded the detectable peptide space; iterative refinement further increased DIA identification while controlling search-space expansion. In DDA-independent reanalysis of Orbitrap human gut metaproteomes, MetaPilot identified 24.4% more peptides than the published DDA-derived library and more than twice the number identified by the matched DDA-assisted workflow. In a timsTOF DIA-PASEF mouse intestinal dataset, MetaPilot outperformed uMetaP in identification depth and enabled genome-resolved functional interpretation. These results establish that DDA-independent DIA metaproteomics, guided by genome-resolved marker evidence, can exceed DDA-assisted workflows in identification depth.

bioinformatics↗

bioRxiv2026

MetaUmbra: Statistically Controlled Genome-Level Presence Inference from Metaproteomic Peptides

Taxonomic interpretation of metaproteomic peptides remains difficult because many peptide sequences are present in proteins from different organisms, reducing taxonomic specificity. Current peptide-centric workflows can report taxonomic summaries or taxon level confidence scores, but they do not provide formal statistical evidence that a taxon is present in the microbiome. Here we present MetaUmbra, a tool that derives genome-level statistical significance values from identified peptides. MetaUmbra builds theoretical peptide lists by in silico digestion of the taxon specific proteins and matches observed peptides against these references. It then combines a conservative significance estimate from unique peptides with a Monte Carlo based p-value for shared peptide evidence estimated under an empirical null model. In the defined community benchmark SIHUMIx, MetaUmbra identified the expected genomes without introducing false-positive genomes after embedding the SIHUMIx genomes in a large gut reference background. In the single strain benchmark Mix24X, all expected genomes were identified with the best statistical significances even after near neighbor and full background expansion. In a hamster gut genome panel, MetaUmbra further preserved an interpretable ranking of candidate genomes in a dense real-data setting. Together, these results show that MetaUmbra can statistically identify the presence of specific microbes in a complex microbiome while maintaining low false-positive calls. MetaUmbra therefore provides a practical framework for converting peptide evidence into genome-level statistical inference in metaproteomics.

bioinformatics↗

bioRxiv2026

ProAR: Probabilistic Autoregressive Modeling for Molecular Dynamics

Understanding the structural dynamics of biomolecules is crucial for uncovering biological functions. As molecular dynamics (MD) simulation data becomes more available, deep generative models have been developed to synthesize realistic MD trajectories. However, existing methods produce fixed-length trajectories by jointly denoising high-dimensional spatiotemporal representations, which conflicts with MDs frame-by-frame integration process and fails to capture time-dependent conformational diversity. Inspired by MDs sequential nature, we introduce a new probabilistic autoregressive (ProAR) framework for trajectory generation. ProAR uses a dual-network system that models each frame as a multivariate Gaussian distribution and employs an anti-drifting sampling strategy to reduce cumulative errors. This approach captures conformational uncertainty and time-coupled structural changes while allowing flexible generation of trajectories of arbitrary length. Experiments on ATLAS, a large-scale protein MD dataset, demonstrate that for long trajectory generation, our model achieves a 7.5% reduction in reconstruction RMSE and an average 25.8% improvement in conformation change accuracy compared to previous state-of-the-art methods. For conformation sampling task, it performs comparably to specialized time-independent models, providing a flexible and dependable alternative to standard MD simulations.

molecular biology↗

bioRxiv2026

Value-Based Evidence Accumulation as a Transdiagnostic Marker of General Distress

General distress cuts across psychiatric symptom domains, yet its computational correlates remain poorly defined. We examined whether drift rate--a core parameter indexing the efficiency of evidence accumulation--is more strongly associated with general distress than with domain-specific symptoms. In a cross-sectional online sample of 441 adults from the general population, participants completed a perceptual and value-based decision-making task, symptom assessments, and cognitive testing. Drift rates were estimated using hierarchical drift-diffusion modeling. Individuals with severe symptom elevations showed robust reductions in drift rate, particularly for value-based decisions. Mixed-effects models demonstrated that general distress, indexed by the Positive Symptom Distress Index, was more strongly associated with value-based than perceptual drift rate, even after accounting for all symptom domains. Value-based drift rate also explained variance in general distress beyond that accounted for by elevated symptoms across domains and selectively attenuated associations with somatization and paranoid symptoms. These findings suggest that value-based evidence accumulation captures a transdiagnostic component of distress-related impairment that is not reducible to symptom burden alone.

pathology↗

bioRxiv2026

Multi-Omics Reveals Activated Fibroblasts with Dual Functional Roles in Repair and Negative Barrier in the CD8+ T Cell Cytotoxic Niche for Lung Cancer Neoadjuvant Therapy

A major challenge in elucidating immune activation and tolerance is that single-omics technologies are inherently limited. Since single-cell transcriptomics lacks spatial information and spatial transcriptomics lacks resolution or depth, neither can adequately model the multidimensional tumor immune microenvironment (TiME) features. We present NICHE (Niche Integrated Cellular Heterogeneity Elucidator), an integrative multi-omics framework. By aligning physical cell-cell interactions with single-cell ligand-receptor (L-R) expression, NICHE leverages single-cell transcriptomics, spatial proteomics, and AI modeling to systematically decode the composition, spatial interactions, and communications within immune functional niches. To validate NICHEs capabilities, we applied it to human tonsil tissue. The framework successfully identified key structural and functional units and deeply elucidated the cell-cell interactions and molecular crosstalk within them. Furthermore, these results were validated through orthogonal methods, including single-cell spatial transcriptomics and multiplexed immunofluorescence, which confirmed the accuracy and reproducibility of our analyses. In non-small cell lung cancer (NSCLC) patients undergoing neoadjuvant immunotherapy, CD8+ T cell cytotoxic niches mediate tumor clearance with variable outcomes, to define the cellular and molecular drivers behind this functional spectrum, we utilized NICHE to analyze the TiME from patients with complete versus partial pathological response, both pre- and post-treatment. We found that neoadjuvant immunotherapy may induce the activation of CXCL12+, NECTIN2+, POSTN+, and COL6A1+ fibroblasts through the activation of cytotoxic T cells. Paradoxically, these fibroblasts support tissue repair but also establish an immunosuppressive niche that protects residual tumor cells, ultimately attenuating treatment efficacy. Our work establishes a multidimensional framework for dissecting dynamic immune activity, with direct implications for understanding tumor immunology and identifying novel therapeutic targets.

immunology↗

bioRxiv2026

Identification of low threshold off-target activation pathways during stimulation of carotid baroreceptor afferents in swine.

ObjectiveElectrical stimulation of the baroreceptors pathways at the carotid sinus bulb - known as baroreflex activation therapy (BAT) - is intended to change autonomic tone and ultimately reduce blood pressure and heart rate. BAT is pre-market approved by the United States Food and Drug Administration (FDA) for the treatment of heart failure and received an FDA humanitarian device exemption for drug resistant hypertension. However, responder rates are limited by side-effects including numbness in the head and neck, altered speech, respiratory constriction, dry cough, vomiting, and altered sensory and motor function of the tongue. We hypothesized that these side-effects are driven by activation of other nearby nerve fibers of similar or lower threshold than the carotid sinus nerve. In this study, we sought to identify the neural sources responsible for off-target muscle activation contributing to these side-effects. These sources would inform strategies mitigating off-target activation in BAT therapy. MethodsDomestic swine were used in this work as the diameter and thickness of the swine carotid artery are closer to human than those of canine models. A monopolar disk electrode mimicking the clinical CVRx(R) Neo electrode was surgically placed proximal to the carotid bifurcation with the position optimized for stimulation dose responsive changes in blood pressure. Evoked responses were recorded during dose response testing from multiple neck muscles, and the corresponding off-target nerve pathways were identified by sequential transection of nearby nerves. ResultsThe following activated off-target muscle groups and their corresponding nerve pathways were verified which included 1) the cricoarytenoid via recurrent laryngeal nerve, 2) cricothyroid via superior laryngeal nerve, 3) sternocleidomastoid via accessory nerve, and 4) the tongue via hypoglossal nerve. The constrictor muscle group was also activated through a more complex neural pathway. ConclusionWe identified multiple sources of therapy-limiting side-effects of BAT in the swine model. These results will help guide the design of improved stimulation electrodes, surgical placement, and parameter programming to reduce BAT-associated side-effects while increasing on-target activation in patients.

bioengineering↗

bioRxiv2025

Dynamic Stiffening to Improve Vasculogenesis of hiPSC-Derived Endothelial Progenitors

Multiple groups have reported on the impact of hydrogel stiffness on vascular network formation in vitro, with overall findings indicating that less stiff hydrogels better support vasculogenesis. However, the majority of this research utilized hydrogels with static stiffness, even though vasculogenesis occurs in tandem with changes in extracellular matrix material properties. To that end, we hypothesize that dynamic modulation of hydrogel stiffness during the process of vasculogenesis, recapitulating changes observed during embryonic development, would improve vascular network formation. Using our Collagen I/Norbornene-modified hyaluronic acid hydrogel system, we swelled in additional crosslinker and photoinitiator and exposed the hydrogel to UV light, enabling hydrogel stiffening at user-defined time points with no significant effect on cell viability. We observed that in situ stiffening at early time points, prior to the onset of significant cell migration, resulted in more robust vascular network formation relative to unstiffened controls, while stiffening at later time points disrupted existing vascular networks. These trends continued in in vivo experiments in nude mice, with cell-laden hydrogels stiffened at early time points resulting in improved blood flow, while those stiffened at later time points had the opposite effect. We hypothesized that this was due to differential impacts of focal adhesion kinase (FAK) activation following in situ stiffening, as supplementation with a Rho kinase inhibitor, downstream of FAK, partially reversed the effects of in situ stiffening. Taken together, this research demonstrates the benefits of incorporating dynamic cues into hydrogel design to create more physiologically relevant vasculature.

bioengineering↗

bioRxiv2025

Unveiling The Origin of Human High-Grade Serous Ovarian Cancer Using Bulk and Single-Nucleus RNA Sequencing

High-grade serous ovarian cancer (HGSOC) is the most lethal gynecologic malignancy, yet its earliest cellular precursors remain incompletely defined. Here, we integrate bulk and single-nucleus RNA sequencing of human fallopian tubes, ovaries, and primary HGSOC tumors to resolve the epithelial hierarchy of the fallopian tube and examine its relationship to malignant states. We identify a rare LGR5/PGR basal population as putative epithelial stem cells and delineate a large pool of OVGP1/RNPC3 progenitors that give rise to both secretory and ciliated lineages. By jointly analyzing normal and malignant transcriptomes, we find that these progenitor populations show the strongest transcriptional and developmental continuity with multiple HGSOC cell states, suggesting that they represent the epithelial compartments most plausibly poised for oncogenic divergence. Transcription factor network reconstruction highlights SPDEF as a key regulator of progenitor identity and NR2F6 as a candidate oncogenic driver, while inferred copy number variations and microenvironmental collagen-integrin signaling further illuminate the genomic and stromal factors that consolidate malignant fate. Together, these findings refine the cellular landscape of the human fallopian tube and identify progenitor-enriched molecular features that could inform earlier detection, improved risk stratification, and new preventive strategies for women at risk of developing HGSOC. One sentence summaryFallopian tube progenitor cells represent a key intermediate state linking normal epithelial hierarchy to the earliest stages of high-grade serous ovarian cancer development.

cancer biology↗

bioRxiv2025

Replicating without stress: a dengue replicon model reveals distinct host rewiring to accommodate persistence

Self-replicating viral RNAs, or replicons, are widely used to study virus-host interactions and screen antiviral compounds, yet their ability to model persistent infection and host adaptation remains poorly understood. Here, we characterize a stable subgenomic dengue virus serotype 2 (DENV2) replicon (DENV2-rep) that supports long-term viral RNA replication in human hepatoma cells without triggering cytotoxicity or canonical stress responses. Compared to live DENV2 infection, which induces ER stress and global translation shutdown, DENV2-rep cells maintain active translation and polysome integrity, while mounting an antiviral transcriptional response and resisting superinfection. Transcriptomic and proteomic profiling reveals that DENV2-rep cells accommodate long-term replication by remodeling of metabolic, secretory, and cytoskeletal pathways. In contrast, acute infection uncouples transcription from protein synthesis, limiting host proteome remodeling. We also detected marked depletion of the NS5 RNA polymerase in the DENV2-rep context. Together, these findings highlight host pathways that can be rewired to support persistent RNA replication and uncover post-translational regulation of viral proteins as a potential vulnerability. Our work clarifies the strengths and limitations of replicons as models for infection and reveals how persistent vRNA replication is tolerated through selective remodeling of translation, organelle composition, and protein stability.

microbiology↗

bioRxiv2025

Nocturnal Navigation via a Time-Compensated Lunar Compass in Bull Ants

Diurnal navigators are well known to orient via the sun, compensating for its movement over time by linking solar position and speed with terrestrial reference points (Beetz & el Jundi, 2018; Dingle, 2014; Dyer, 1987; Mouritsen & Frost, 2002; Stalleicken et al., 2005; Wehner & Lanfranconi, 1981; Wehner & Muller, 1993). In contrast to the sun, the moon is not only dimmer, but its presence is also highly variable, both temporally and in its illuminance, making it a more complex cue to predict. Yet nocturnal animals employ a celestial compass to orient at night (i.e. the moon and stars; (Dacke et al., 2003, 2013; Dreyer et al., 2025), but navigation via a time-compensated lunar compass has yet to be demonstrated. Nocturnal bull ants (Myrmecia midas) navigate home overnight under the dimmest of visual conditions, relying in part on a nocturnal path-integration system informed by lunar cues (Freas et al., 2024). Here, we show the first evidence of a time-compensated lunar compass in a homing animal, with path-integrating ants adjusting path headings by predicting the lunar azimuthal position throughout the night. Ants predict the moons future position based upon a generalised internal estimate of the lunar ephemeris. Rather than detailed knowledge of the moons nightly schedule, these ants form a lunar path representation via a speed-step function, reflecting the moons accelerating then decelerating arc. However, these ants also predictably under- or overestimate the shape of this arc based on individual experience, suggesting a level of temporal uncertainty regarding when the moon rapidly shifts from the eastern to western sky. Additionally, updating this lunar compass relies on the presence of a directionally informative skyline, which allows occasional cross-referencing of lunar movement with external fixed compass cues near the horizon.

ecology↗

bioRxiv2025

Bona fide hematopoietic stem cells in zebrafish originate from the supra-intestinal artery

Hematopoietic stem cells (HSCs) in vertebrates are widely believed to originate from hemogenic endothelial cells (HECs) in the dorsal aorta (DA) through endothelial-to-hematopoietic transition. However, conclusive in vivo DA-specific lineage-tracing studies to confirm this origin are lacking. Here, leveraging infrared light-mediated temporospatial-resolution lineage-tracing technology, we demonstrate that in zebrafish, HSCs do not originate from the DA. Instead, following the DA hematopoiesis, which produces hematopoietic precursors with limited renewal potential, lifelong renewable HSCs emerge from the supra-intestinal artery (SIA). Remarkably, while the DA produces hematopoietic cells from unipotent HECs, the SIA generates hematopoietic cells from both unipotent HECs and bipotent vessel-resident hemangioblasts (VRHs). These VRHs undergo multiple rounds of division and budding, generating both hematopoietic cells and SIA-resident endothelial cells. The distinct characteristics of DA and SIA cells are further reinforced by single-cell transcriptomic profiling. Our findings challenge the current paradigm of HSC development, underscoring the complexity of vertebrate hematopoiesis.

developmental biology↗

bioRxiv2025

Zebrafish armc9 mutant reveals a link between ARM-Type fold domain loss, inflammation, and spinal curvature

The vertebral column (VC) defines the central axis of vertebrates. Among its malformations, adolescent idiopathic scoliosis (AIS) is the most common, characterized by lateral spine curvature without congenital abnormalities. While its etiology remains elusive, genetic studies in zebrafish have linked AIS to skeletogenesis, cilia structure, cerebrospinal fluid flow, and inflammation. In this study, we characterized the zebrafish armc9 mutant, exhibiting AIS-like curvature without vertebral malformations, subcommissural organ, or Reissner fiber structure disruptions. We also detected an inflammatory response at the curvature origin. Structural bioinformatics and molecular docking analyses suggest that the mutant Armc9 protein partially lacks the functional ARM-type fold domain, which is crucial for its interaction with Togaram1, revealing that both proteins are relevant for ciliary signaling and spine curvature. Phylogenetic analyses indicate that the ARMC gene family encompasses 12 members, highlighting that the Armc9 gene has deep evolutionary roots. Our results demonstrate that the zebrafish armc9 mutant accurately represents the AIS phenotype, suggesting that the loss of Armc9s C-terminal ARM-type fold domain could affect the ciliary integrity and contribute to spine curvature by altering the skeletogenic environment. Taken together, this study proposes that the zebrafish armc9 mutant offers a unique opportunity to understand the etiology of AIS and provides new insights into the molecular interaction networks of Armc9 in ciliary signaling.

developmental biology↗

bioRxiv2025

Diurnal Bull Ants Navigating by Moonlight: Polarised Light Homing in Myrmecia tarsata

We present evidence that the faint polarised moonlight pattern of the sky can be detected and used for navigation in the diurnal bull ant Myrmecia tarsata, despite this species lacking the highly refined low-light visual specialisations of the nocturnal bull ant, Myrmecia midas. The position of celestial bodies such as the sun and moon can provide navigating animals directional information, yet direct observation can often be occluded. Animals can estimate positional information of solar and lunar cues via their polarised light pattern, present across the sky. The suns polarisation pattern is widely used in animals and a similar, yet much fainter pattern is produced by the moon, yet it is unknown how widespread moonlights use is in navigating animals. Here, we demonstrate that the bull ant Myrmecia tarsata, which forages throughout the day, returning home at sunset, can use both solar and lunar polarised light patterns to navigate. We compare these findings to the closely related M. midas navigating under identical light conditions, as this nocturnal bull ant is known to rely on these polarised light patterns as part of their celestial compass. While M. midas and M. tarsata can clearly use both the solar and lunar polarised light patterns to navigate, M. tarsata showed degraded performance under polarised moonlight as a function of lunar phase, decreasing performance as illumination decreased. M. midas in contrast, exhibited impressive attendance to the overhead lunar polarisation pattern throughout the lunar month, illustrating the highly specialised low-light-detection adaptations of bull ants.

animal behavior and cognition↗

bioRxiv2025

Intense oocyte competition builds the female reproductive reserve in mice

During the process of ovariogenesis, more than two-thirds of germ cells are sacrificed to enhance the quality of the remaining oocytes. But the detail process of selection is not completely understood in mammals. Here, by creating a high resolution 4- dimensional ovariogenesis imaging system, we recorded the progress of oocyte fate determination in live mouse ovaries, and discovered an oocyte competition mechanism that determines the survival of oocytes. We found that dominant oocytes catch and absorb the cell debris from sacrificed oocytes to enrich their cytoplasm for survival. Single-cell sequencing demonstrated that this competition follows a classic autophagy- driven model. Blocking competition by suppressing autophagy resulted in an enlarged pool of survived oocytes. However, these oocytes lacking competition were unable to fully develop and contribute to fertility. Our study suggests that mammals have evolved a unique intensive competition system to enhance oocyte quality, necessary to support their long reproductive lifespan.

developmental biology↗

bioRxiv2025

Multi-Region Brain Organoid: Fusion Organoid with Cerebral, Endothelial and Mid-Hindbrain Components

Brain organoid technology has revolutionized our ability to model human neurodevelopment in vitro. However, current techniques remain limited by their reliance on simplified endothelial cell populations rather than a complete endothelial system. We engineered Multi-Region Brain Organoids (MRBOs) that integrate cerebral, mid/hindbrain, and complex endothelial organoids into one structure. Different from the earlier approaches based on isolated HUVECs, our endothelial organoids contain diverse vascular cell types, including vascular progenitors, mature endothelial cells, pericytes, proliferating angiogenic cells and stromal cells. Our strategy employs a sequential modulation of key developmental pathways to generate individual organoids, followed by optimized fusion conditions that maintain regional identities while supporting cellular integration. Single-nucleus RNA sequencing shows that MRBOs develop discrete neural populations specific to each brain region alongside specialized endothelial populations that establish paracrine signalling networks. Integration analysis with human fetal brain data shows that MRBOs contribute to 80% of cellular clusters found in human fetal brain tissue (Carnegie stages 12-16), whereas CellChat analysis identifies 13 previously uncharacterized endothelial-neural signalling interactions. Notably, we uncover endothelial-derived factors that support the persistence of intermediate progenitor populations during hindbrain development, but not during the cerebral one thereby revealing a new role for complex endothelial populations in regional brain patterning. This platform enables matching multiple developmental regions, fully incorporating the endothelial nature at the same time providing opportunities for studying neurodevelopmental disorders in which neural-endothelial interactions are disrupted. Our engineered MRBO system establishes a foundation for investigating complex neurodevelopmental processes, providing an enabling context closer to physiological relevance.

bioengineering↗

bioRxiv2025

Ants oscillate and scan more in navigation when the visual scene changes

Solitarily foraging ants learn to navigate between important locations by comparing their current view with memorized scenes along a familiar route. As desert ants, in particular, travel between their nest and a food source, they establish stable and visually guided routes that guide them without relying on trail pheromones. We investigated how changes in familiar visual scenes affect the navigation of the red honey ant (Melophorus bagoti). In Experiment 1, ants were trained to follow a one-way diamond-shaped path to forage and return home. We manipulated scene familiarity by adding a board on their homebound route just before the nest. In Experiment 2, ants were trained to travel a straight path from their nest to a feeder, and we removed the prominent landmarks on the route after they had established a stable route. We predicted that these scene changes would cause the ants to deviate from their usual straight paths, slow down, scan more, and increase their lateral oscillations to gather additional information. Our findings showed that when the familiar scene was changed, ants oscillated more, slowed their speed, and increased scanning bouts, indicating a shift from exploiting known information to more actively exploring and learning new visual cues. These results suggest that scene familiarity plays a crucial role in ant navigation, and changes in their visual environment lead to distinct behavioral adaptations aimed at learning about the new cues.

ecology↗