Search Results (1,023)

Hydrogen sulfide (H₂S) is a vital gasotransmitter essential for maintaining redox homeostasis and modulating inflammatory responses. Herein, we examined a collection of Drosophila melanogaster knockout (KO) lines generated in our laboratory, lacking key genes involved in transsulfuration (cbs, cse) and sulfide metabolism (tst1), to elucidate the role of this adaptive system in immunity. Genetic ablation of these pathways results in profound H₂S deficiency and hyperhomocysteinemia, leading to chronic oxidative stress. This leads to constitutive activation of major immune pathways—including IMD, Toll, and JAK-STAT—even in the absence of infection, a hallmark of chronic inflammation. Indeed, transcriptomic analysis and qRT-PCR studies revealed significant upregulation of these pathways in double (cbs; cse) and triple (cbs; cse; tst1) KO flies under control conditions. Following septic injury with Bacillus subtilis, double and triple KO flies exhibited increased expression of antimicrobial peptides (AMPs) and pattern recognition receptors compared to control and TST1-/- single KO lines. Notably, double and triple KO lines showed more prolonged immune activation. The triple KO flies exhibited the worst survival rate following septic injury. These results demonstrate that H₂S deficiency causes chronic inflammation through the sustained activation of immune pathways, highlighting sulfur metabolism as a crucial regulator of homeostasis and innate immunity in Drosophila. Full article

Across animals, variation in adult body size is accompanied by coordinated variation in the size of individual morphological traits. However, the same morphological trait can scale differently with body size depending on what drives the size variation. In Drosophila melanogaster, for example, wing size scales differently from body size when size varies because of developmental nutrition versus developmental temperature. Whether the genetic basis of size plasticity and scaling is shared across different environmental regulators of size remains unclear, but it is central to predicting how selection acts on the developmental mechanisms that regulate trait size, plasticity, and morphological scaling. Using ~100 isogenic D. melanogaster lineages, we measured wing and leg size across nutritional and thermal treatments. For each lineage, we estimated nutritional and thermal plasticity for both traits, as well as the wing–leg individual-level scaling relationship, or ILSR, generated by each environmental source of size variation. We found extensive genetic variation in both thermal and nutritional plasticity for wings and legs and in the slope of the ILSR between them. However, a lineage’s thermal plasticity was genetically uncorrelated with its nutritional plasticity for either trait, and we detected no genetic correlation between the slopes of thermal and nutritional wing–leg ILSRs. We also found no genetic correlation in the slope of nutritional wing–leg ILSRs between temperatures. Thus, the slope of a lineage’s nutritional ILSR at 17 °C was not predictive of its slope at 25 °C or 28 °C. Nevertheless, the overall pattern of nutritional ILSRs was conserved across temperatures. These results suggest that the genetic architecture of size plasticity and scaling depends on the environmental source of size variation. Consequently, the evolutionary response of scaling to selection in heterogeneous environments may not be predictable from genetic variation measured in any single environment. Full article

Glycyrrhiza glabra leaves (GGL) are an underutilized by-product of the licorice supply chain and a valuable source of bioactive flavanones. In this study, supercritical CO₂ fluid extraction (SFE-CO₂) was optimized for the selective recovery of leaf-exudate flavanones (pinocembrin, licoflavanone, and glabranin). Response surface methodology combined with UHPLC–UV profiling enabled the identification of optimal conditions, yielding high-purity (31% flavanones), solvent-free, and ready-to-use enriched extracts. SFE-CO₂ outperformed solvent-based liquid extraction in terms of selectivity and enrichment capacity. Path2Green assessment indicated an environmentally sustainable and scalable profile. The extract exhibited antioxidant and anti-aging effects in a yeast model of Parkinson’s disease (PD), extending lifespan and reducing oxidative stress. Pinocembrin, the most abundant flavanone, inhibited α-synuclein aggregation in vitro and ameliorated PD-related phenotypes in a Drosophila melanogaster model, improving locomotor performance and increasing cellular energy metabolism. These results support the sustainable valorization of GGL into flavanone-enriched extracts with potential nutraceutical applications for aging-related disorders. Full article

Background/Objectives: The study sought to investigate the neurotoxicity of NBOHs, which are a class of new psychoactive substances (NPSs) that act as agonists of serotonin 5-HT2A and 5-HT2C receptors, resulting in hallucinogenic effects similar to those of LSD, often found in paper blotters. Methods: Differentiated SH-SY5Y cells were incubated for 24 h with 0.5, 1.0, 3.5, 5 and 10 nM of the derivatives 25I-NBOH, 25E-NBOH and 25B-NBOH. Cell viability was assessed by MTT reduction and neutral red uptake assays. Mitochondrial membrane potential and production of free radicals were also evaluated. In addition, Drosophila melanogaster flies were exposed to 5, 10 and 50 nM of each NBOH derivative for 4 h. Negative geotaxis, CAT, and AChE enzymatic activities were evaluated. Results: Both 25E-NBOH and 25B-NBOH decreased cell viability. Furthermore, mitochondrial hyperpolarization of cells and an increase in oxidative species were observed in response to higher concentrations of 25E-NBOH and 25B-NBOH. In D. melanogaster, 25B-NBOH increased climbing time and both 25E-NBOH and 25B-NBOH increased CAT and AChE activities at 50 nM. Conclusions: These findings indicate greater toxicity of the 25E- and 25B-NBOH compared to 25I-NBOH, suggesting metabolic activity or mitochondrial signaling, oxidative stress, and cholinergic dysregulation as a potential mechanism related to NBOH neurotoxicity. Full article

RNA N⁶-methyladenosine (m⁶A) modification is a pivotal post-transcriptional regulator of diverse biological processes. Despite the growing interest in insect epitranscriptomics, a systematic evaluation of research trends and hotspots remains lacking. Here, we conducted a bibliometric analysis to map the global landscape of METTL3 and m⁶A research in entomology over the past decade. Our results reveal a steady increase in publications, signaling a phase of rapid expansion in this field. Notably, the number of studies on METTL3 is significantly lower than that of general m⁶A research, suggesting that current efforts prioritize phenotypic over the mechanistic roles of core regulatory components. Keyword co-occurrence analysis identifies Bombyx mori, Locusta migratoria, and Drosophila melanogaster as the primary model systems. Research hotspots predominantly center on METTL3-mediated regulation of development, behavioral plasticity, immunity, and host–pathogen interactions. These findings highlight insect METTL3 as a burgeoning research frontier. Future studies should emphasize cross-species comparisons and the systematic dissection of regulatory networks to provide novel theoretical frameworks and molecular targets for sustainable pest management and resource insect utilization. Full article

Vitamin K is a well-established cofactor for γ-glutamyl carboxylase in vertebrates, yet its physiological functions in insects remain poorly understood. Warfarin, a vitamin K antagonist, inhibits vitamin K epoxide reductase, thereby blocking the vitamin K cycle. We have previously demonstrated that, in Drosophila melanogaster, vitamin K3 (menadione sodium bisulfite, MSB) is converted into endogenous vitamin K2, and that warfarin interferes with this cycle, leading to reduced oxidative stress. In the present study, we investigated the effects of chronic exposure to MSB (3.5 mM) and/or warfarin (1 mM and 10 mM) during larval development on adult survival, lifespan, mitochondrial ATP hydrolysis rate, and the transcriptional expression of target genes. Warfarin at 1 mM did not affect development or egg laying, whereas 10 mM caused significant postembryonic lethality. Transcriptional analysis revealed marked sex-specific effects. In females, warfarin downregulated the levy gene (encoding a complex IV subunit) and the ATPsynB gene (encoding an ATP synthase subunit B), while MSB suppressed cell cycle genes (cmet, sti, mcm2). In contrast, males exhibited upregulation of COQ7 (encoding a key enzyme in ubiquinone biosynthesis) in response to warfarin and increased sti expression in response to MSB, suggesting a more robust compensatory response. Notably, the combined application of MSB and warfarin increased mitochondrial ATP hydrolysis, thereby offsetting the negative physiological effects despite persistent transcriptional suppression. In females, either MSB or warfarin alone significantly reduced median lifespan; however, when applied together, female lifespan was restored to control levels. In males, the effects were considerably weaker. Thus, this study reveals a novel, sex-specific interaction between vitamin K and its antagonist in D. melanogaster, providing new insights into mitochondrial adaptation and sexual dimorphism in metabolic stress responses. Full article

Gustatory perception in honeybees is a key determinant of foraging decisions and pollen source selection. However, the molecular mechanisms underlying this sensory discrimination remain poorly understood. To investigate these mechanisms during the collection of pollen from different floral sources, this study utilized antennae from worker bees foraging on pear and rapeseed pollen, and non-pollen-foraging workers as controls. Illumina high-throughput transcriptome sequencing was employed to identify differentially expressed genes (DEGs), perform functional annotation, and characterize gustatory receptor (GR) genes. Compared with the control group, 583 DEGs and 516 DEGs were identified in pear-pollen and rapeseed-pollen foragers, respectively, whereas only 73 DEGs were detected between the two pollen-foraging groups. Several DEGs were associated with chemosensory perception, signal transduction, energy metabolism, and immune responses. Notably, genes involved in membrane-associated signaling and stimulus response exhibited differential expression patterns among foraging groups, suggesting adaptive molecular responses to distinct floral resources. Gene Ontology (GO) analysis indicated that DEGs were primarily associated with cellular processes, membrane components, and binding functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment highlighted significant involvement in phagosome, phosphatidylinositol signaling system, oxidative phosphorylation, and extracellular matrix–receptor interaction. Notably, seven GR-related genes were identified in the antennal transcriptome, including five known GR genes and two novel candidates, all with complete open reading frames. Four of these genes featured the canonical seven-transmembrane domain structure of insect GRs. Phylogenetic analysis, in addition to the known sugar receptors AmelGR43a, AmelGR64f, and AmelGR64f-X1, based on GRs from Apis mellifera and Drosophila melanogaster suggested that AmelGR28b, AmelGR10, AmelGR12, and AmelGR13 may belong to the bitter taste receptor family. Quantitative real-time PCR (qRT-PCR) validation demonstrated that the expression patterns of the selected seven DEGs were consistent with the RNA-seq results. This study reveals differential expression patterns and potential functional divergence of gustatory receptor genes in Apis mellifera during pollen collection from different floral sources. It provides important molecular evidence for understanding how honeybees accurately recognize and preferentially forage specific pollen sources via gustatory perception, and offers valuable theoretical and practical insights for honeybee behavioral ecology and crop pollination management. Full article

Background: Accumulating evidence suggests that environmental light cues influence brain function and neurodegenerative processes; however, the underlying cellular mechanisms remain incompletely understood. Methods: Here, using a Tau-overexpressing Drosophila model, we investigated how light exposure modulates neurodegeneration-associated phenotypes, with a particular focus on adenosine signaling and mitochondrial homeostasis. We performed behavioral assays, biochemical measurements, genetic interference targeting the adenosine receptor, and mito-QC reporter analysis to assess mitochondrial quality control. Results: We show that light exposure ameliorates Tau-induced behavioral impairments and neuropathological features, reducing climbing time by approximately 29% in males and 45% in females, and extending median lifespan by ~29% in males and ~26% in females. Notably, biochemical analyses revealed that light exposure significantly increases brain adenosine levels at ZT12 by approximately 5 to 6 nmol/L in both sexes (p < 0.01), suggesting a light-dependent modulation of adenosine availability. To further examine the role of adenosine signaling, we performed genetic interference experiments targeting the adenosine receptor. These results indicate that adenosine receptor-associated signaling is functionally involved in the beneficial effects of light, as disruption of this pathway attenuates the light-induced improvements in behavioral and mitochondrial phenotypes. Using a mito-QC reporter system, we further show that light exposure enhances mitochondrial quality control, as reflected by a ~2.3-fold increase in mitolysosome density (p < 0.001). Importantly, this effect is modulated by the functional state of adenosine signaling, suggesting a potential interaction between these processes. Conclusions: Together, our findings indicate that light exposure is associated with coordinated changes in adenosine signaling and mitochondrial quality control, which may contribute to the attenuation of Tau-induced deficits in Drosophila. This work provides insight into how environmental light cues may influence neurodegeneration-related cellular processes and highlights the potential relevance of light-based interventions for future mechanistic and translational studies. Full article

Drosophila melanogaster, widely known as the fruit fly, has emerged as a pivotal model organism for studying development and signaling transduction. Its fully sequenced genome, short generation time, and powerful genetic toolkit—including the Gal4/UAS system, RNA interference, and CRISPR-Cas9—enable precise, tissue-specific manipulation and high-throughput functional analyses. Despite differences in anatomy, the internal organ systems of Drosophila melanogaster, including the nervous system, heart, fat body, oenocytes, and nephrocytes, exhibit conserved molecular pathways and physiological functions comparable to those of humans. The morphological differences between invertebrates and vertebrates have long led researchers to undervalue the studies of insects in underlying the pathogenesis of human diseases. Over the past decades, the fruit fly has been widely validated for modeling the pathogenesis of neurodegenerative, cardiovascular, metabolic, renal, and muscular disorders. In this review, we systematically summarize the conserved molecular pathways and organ functions between the fruit fly and human, and provide examples of recent studies that use the fruit fly as a model system to answer questions associated with human diseases. We also discuss how Drosophila help researchers to fulfill the gap from mechanistic study toward translational research, and provide methodological considerations regarding the utility of Drosophila models in drug screening. Full article

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by β-amyloid (Aβ) accumulation and a breakdown of mitochondrial homeostasis. Vitamin K2 (VK2) has emerged as a potential neuroprotective agent, yet the specific molecular cascades linking its intervention to the restoration of mitochondrial integrity remain poorly understood. This study utilizes an AD Drosophila model to investigate the efficacy of VK2 and elucidates its multidimensional regulatory mechanisms. Behavioral analysis showed that VK2 significantly rescued locomotor impairments, improving both vertical climbing and horizontal walking performance. Crucially, VK2 intervention achieved a systemic rescue of mitochondrial health: transmission electron microscopy (TEM) confirmed the preservation of mitochondrial ultrastructure and cristae density, while biochemical assays demonstrated a robust recovery of bioenergetic markers, including ATP levels and the NAD⁺/NADH ratio. Furthermore, VK2 treatment stabilized the mitochondrial membrane potential (MMP) and effectively attenuated the accumulation of reactive oxygen species (ROS). To identify the molecular drivers of this recovery, an unbiased integration of human clinical transcriptomic data and network pharmacology prioritized the EGFR-Ras-ERK signaling axis as a central hub. In vivo validation confirmed that VK2 suppresses the pathological overactivation of this cascade. VK2 reduced EGFR phosphorylation in parallel with the effects observed for the EGFR inhibitor Gefitinib. Collectively, our findings show that VK2 ameliorates locomotor deficits and mitochondrial dysfunction in Aβ42-expressing flies and that these effects are associated with suppression of the EGFR-Ras-ERK signaling axis. Further studies are required to establish direct target engagement and pathway causality. Full article

Functional changes in PNPLA6 (Patatin-like phospholipase domain-containing protein 6), caused by gene mutations or inhibition by organophosphates, affect the levels of various phospholipids. In humans, this leads to organophosphorus compound-induced delayed neurotoxicity syndrome (OPIDN) and a number of rare diseases. In this study, we analyze the role of the Swiss cheese gene (sws), an ortholog of PNPLA6, in spermatogenesis in Drosophila melanogaster. We report that the sws¹ mutation affects membrane remodeling during spermatid individualization, as well as spermatid coiling during the late stages of spermatogenesis. In addition, the sws¹ mutation leads to changes in the transcriptome in the testes of flies. We also demonstrate that sws is required for the proper functioning of important biological barriers in Drosophila. Full article

Analysis of freely moving Drosophila captures complex movement behaviors. However, previous experiments have been limited by the inability to distinguish between falls and downward jumps (downjumps). Here, individual flies moving freely in a culture vial were recorded using a single 58 fps video camera. Upward jumps were readily identified by positive movement in the vertical direction. Several statistical and machine learning methods were used to distinguish between falls and downjumps, including Principal Component Analysis (PCA), K-Means Clustering, Uniform Manifold Approximation and Projection (UMAP), Hierarchical Density-Based Spatial Clustering with Applications to Noise (HDBSCAN) and Shapley Additive Explanations (SHAP). Falls were abundant and characterized by an initial velocity consistent with simple acceleration due to gravity. Downjumps were more rare, and were characterized by a greater initial velocity, indicating active propulsion by the fly. Aged flies took longer to resume movement after a fall, suggesting possible negative effects of falls. Falls in young w[1118]-strain flies exhibited mid-event velocities that were lower than expected, indicating some compensatory behavior that was reduced in aged flies. These methods should facilitate future studies of the effects of aging and neurodegenerative disease models on locomotor behaviors and falls, including the testing of potential interventions. Full article

FLASH RT, which employs ultra-high dose rates (UHDR), has shown potential in reducing irradiation-induced damage to normal tissue while maintaining effective tumor targeting. For successful clinical translation, mechanistic explanation behind the so-called FLASH effect has yet to be deciphered and new in vivo model systems for mechanistic studies are therefore of high demand. We investigated the differential effects of UHDR (3000–7000 Gy/s) and conventional (CONV) (0.5 Gy/s) irradiation in D. melanogaster by irradiating adult female flies with 16 MeV and 9 MeV electron beams using an adapted clinical linear accelerator. Substantial lifespan prolongations were observed in single high-dose UHDR-irradiated groups compared to CONV irradiation groups with increasing doses (1000–1500 Gy). Split-dose UHDR irradiation further increased the lifespan compared to single high-dose UHDR irradiation. In addition, climbing deficits were induced by 300 Gy of CONV irradiation, but not by single high-dose UHDR irradiation. Additionally, we identified increased levels of lipid peroxidation in D. melanogaster brains indicating ferroptosis following CONV irradiation, which was not observed after single high-dose UHDR irradiation. Using relevant biological endpoints, we here demonstrate D. melanogaster with its advantageous characteristics to be a highly practical preclinical model organism to mechanistically investigate differential responses to UHDR and CONV irradiation. Full article

Reproductive performance is closely linked to nutrient availability, yet the specific nutrient-derived cues governing oviposition decision-making remain inadequately defined. Chemically defined holidic media used in Drosophila melanogaster studies provide precise control over dietary components; however, the mechanisms by which individual nutrients exert acute effects on inter-organ endocrine signaling to trigger oviposition behavior are not well understood. In a two-choice oviposition assay, where substrates are presented within the same chamber, we observed that HM induces a rapid increase in egg laying in D. melanogaster compared to grape/apple juice media (GJM), yielding 4.02× and 5.27× more eggs at 2 h and 8 h, respectively. Systematic nutrient omission and subsequent supplementation identify lysine as the key essential amino acid driving this rapid oviposition response. Notably, this phenomenon reflects a short-term oviposition reaction, rather than long-term nutritional modulation of ovarian development. Lysine supplementation results in elevated levels of 20-hydroxyecdysone (20E) and juvenile hormone (JH), as well as the induction of ecdysis-triggering hormone (ETH) expression. Pharmacological interventions support a model in which a 20E–ETH–JH endocrine cascade underlies the lysine-dependent phenotype: methoprene phenocopies the effect, whereas precocene suppresses the HM-induced increase in egg laying. A 20E analog similarly enhances oviposition. Consistent with the role of JH in reproductive tissues, activation of the JH pathway correlates with increased expression of extracellular matrix genes in the ovarian muscle. Collectively, these results demonstrate that lysine acts as a critical nutritional cue to activate the 20E–ETH–JH endocrine axis and acutely promote egg laying behavior in D. melanogaster over a short time period. This provides a defined framework for investigating the short-term nutrient hormone regulation of ovulation and offers a practical strategy for efficient egg collection. Full article

Environmental pollution has emerged as one of the most significant threats to human and ecosystem health, with growing evidence suggesting that chronic exposure to toxic substances may accelerate aging. The concept of gerontogens, toxic compounds capable of accelerating this biological process, has gained increasing attention in toxicological research, particularly in the context of global demographic shifts toward older populations. Current research on gerontogens relies heavily on invertebrate models with short lifespans, such as Caenorhabditis elegans, Drosophila melanogaster, and Saccharomyces cerevisiae, which are valuable for studying conserved mechanisms in aging pathways, but present significant limitations for translational accuracy to many aspects of vertebrate biology. Vertebrate models traditionally employed in toxicology, including mice and zebrafish, require substantially longer experimental timelines and higher financial investments, making lifetime exposure and aging assays particularly challenging. In this context, the turquoise killifish Nothobranchius furzeri emerges as a highly promising vertebrate model for aging toxicology research. Recognized as the shortest-lived vertebrate species maintained under laboratory conditions, N. furzeri reaches sexual maturity within 14 days and displays complete senescence by 4 months of age, at which point individuals are considered elderly, offering a decisive advantage over conventional vertebrate models. Furthermore, its capacity for embryonic diapause enables practical embryo storage, long-distance transport, and synchronized hatching, greatly facilitating experimental designs. Although N. furzeri is well established in gerontological research, with studies addressing hallmarks of aging such as telomere shortening, neurodegeneration, and cellular senescence, its application in ecotoxicology remains remarkably limited, with fewer than 10 published studies to date. This article argues that N. furzeri may represent a critical bridge between toxicology and aging research, offering an efficient and translationally relevant platform for investigating the effects of environmental contaminants on vertebrate aging. Current limitations of the model, such as lack of husbandry standardization, are also discussed. Expanding its use in this field holds considerable potential for advancing evidence-based strategies in public health and environmental conservation related to chronic exposure to contaminants. Full article