STAR Protocols
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2022
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IF 0
Yajie Gao,Yuanxiang Zhu,Qinmiao Sun,Dahua Chen
Summary Here, we provide an optimized RNA-induced silencing complex (RISC) assembly and cleavage protocol in vitro without using radiolabeled RNA. The protocol is useful to characterize the biochemical properties of the RISC. We describe the preparation of RNA probes, the target RNA, and Drosophila cell lysates for RISC assembly assay. We then detail AGO1 complexes immunoprecipitation for RISC cleavage assay. This protocol can detect RISC assembly and cleavage products within 5 days. Moreover, it can detect 5′- and 3′-cleavage products simultaneously. For complete details on the use and execution of this protocol, please refer to Gao et al. (2022) .
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PEST MANAGEMENT SCIENCE
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2026
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IF 4.2
Fan Zhang,Yu-Qiu Xie,Yan-Chao Zhang,Cong-Fen Gao
BACKGROUND Moulting is a crucial process in the growth and development of insects and is also a key focus in the development of insecticides. Cyromazine, a moulting disruptor, is widely used to control dipteran pests, yet its mechanism of action and the genetic basis of resistance remain unclear. This study aimed to systematically investigate the inheritance pattern, potential resistance mechanisms, and associated fitness costs by developing a cyromazine-resistant Drosophila melanogaster strain through continuous laboratory selection. RESULTS After long-term directional selection, we successfully established the cyromazine-resistance (Cyr-R) strain with a resistance ratio of 50.19-fold. Genetic analysis revealed that the resistance trait is autosomal, incompletely dominance and polygenic. The Cyr-R strain exhibited only a 4.64-fold low level cross-resistance to chlorfluazuron, with no cross-resistance to other tested insect growth regulators. Notably, the Cyr-R strain exhibited substantial fitness costs: the fecundity and egg hatch rate, pupation rate and eclosion rate had significantly reduced. RNA-sequencing and enzyme activity assays combined with synergism bioassays demonstrated that the cytochrome P450 enzymes play crucial roles in resistance to cyromazine. Quantitative real-time polymerase chain reaction (qRT-PCR) and molecular docking suggested the possibility that the binding affinities between cyromazine and four cytochrome P450 monooxygenase (P450) proteins. CONCLUSION Our study has provided comprehensive characterizations of cyromazine resistance in D. melanogaster , highlighting its slow resistance development, limited cross-resistance, and fitness costs. These findings offer valuable insights for resistance management strategies and further mechanistic studies. © 2026 Society of Chemical Industry.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Hailiang Guo,Wenjin Xia,Changyang Chen,Song Yang,Dongmei Sun,Jizu Qiao,Jianya Su
Nicotinamidase (Naam), which is essential for insect NAD synthesis but absent in mammals, represents a potential pesticide target. Virtual screening identified 1-[4-(trifluoromethyl)-3-pyridinyl]-ethanone (4-TFPE) as a potent inhibitor of Myzus persicae (IC50 = 4.156 μM) and Drosophila melanogaster (IC50 = 2.855 μM) Naam. While 4-TFPE reduced pupal weight, development, and longevity in both species, it showed no lethal toxicity. Crucially, Naam expression was upregulated by nicotinamide accumulation, revealing a feedback regulation mechanism that compromises its pesticidal potential. Furthermore, Flumetnicam’s neurotoxic effects were found to be Naam-independent. These findings highlight how target compensation mechanisms can undermine pesticide development focused on single metabolic enzymes, suggesting the need for multitarget strategies.
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PHYTOMEDICINE
·
2026
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IF 11.3
Rong Xu,Yingna Li,Xinyue Chen,Peiyao Gao,Chunlei Ji,Yiming Zhao,Chuang Xue,Hui Li,Changjiu Cai,Fangbing Liu,Liwei Sun
Background Sleep deprivation is closely associated with metabolic dysregulation, yet effective preventive interventions remain limited. Although Rhodiola rosea ( R. rosea ) exhibits promising sleep-modulating potential, the specific active components mediating its effects and the underlying mechanisms remain poorly elucidated. Purpose This study aimed to identify the sleep-regulatory component of R. rosea and elucidate its molecular mechanism. Methods A bioactivity-guided fractionation strategy was applied using a sleep-deprived Drosophila melanogaster model. The active fraction was purified and structurally characterized by chromatographic separation and spectroscopic analyses. Sleep behavior, circadian rhythmicity, oxidative stress indices, neurotransmitter levels, lipid metabolic parameters, and brain transcriptomic profiles were systematically evaluated. Mechanistic validation was performed using neuronal genetic manipulation and dual-luciferase reporter assays targeting lipid metabolism-related pathways. Results An acidic glycopeptide fraction derived from R. rosea (WRPA, 76 kDa) markedly increased total sleep duration by approximately 20%, reduced sleep fragmentation by approximately 20%, and improved circadian stability, locomotor activity, and cognitive performance in sleep-deprived flies. WRPA significantly alleviated oxidative stress by reducing reactive oxygen species (ROS) and malondialdehyde (MDA) levels and restoring antioxidant capacity, while normalizing gamma-aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT) levels in the brain. Transcriptomic analysis revealed lipid metabolism as the primary pathway modulated by WRPA, with acetyl-CoA carboxylase (ACC) identified as a key target. WRPA suppressed ACC expression by inhibiting sterol regulatory element-binding protein (SREBP)-mediated transcription, leading to decreased triglyceride, cholesterol, and free fatty acid accumulation. Importantly, WRPA effectively rescued sleep disturbances induced by neuronal ACC overexpression, confirming an SREBP/ACC-dependent mechanism. Conclusion WRPA is identified as a novel sleep-regulatory acidic glycopeptide derived from R. rosea , which restores sleep homeostasis by maintaining lipid metabolic stability through the SREBP-ACC signaling axis. These findings expand the pharmacological repertoire of R. rosea beyond small molecules and highlight glycopeptides as promising lead candidates for metabolic sleep disorder intervention.
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Frontiers in Microbiology
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2026
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IF 5.8
Yuling Dong,Beibei Du,Changjian Xie,Shujing Zhang,Qiuxiang Pang,Desheng Zhang
Potassium sorbate (PS) is a widely used antimicrobial additive employed as a preservative in food, cosmetics, and animal feed. Early childhood represents a critical developmental window characterized by rapid growth, immune system maturation, gut microbiota establishment, and physiological homeostasis development, which may be particularly vulnerable to chemical exposures (during which chemical exposures may exert heightened impacts). Nevertheless, the effects of PS on childhood development and gut microbiota remain poorly understood, and its potential trans-generational effects have yet to be elucidated. In this study, we employed Drosophila melanogaster, an established model for studying evolutionarily conserved aspects of development, metabolism, and host–microbiota interactions, to systematically evaluate PS toxicity across multiple parameters: developmental dynamics, gut microbiota composition, gene regulation in adulthood, and trans-generational effects. Our findings demonstrate a dose-dependent biphasic response: while low-dose PS exposure (25 mg/L) accelerated larval pupation and adult emergence, suggesting a potential growth-promoting effect, high-dose exposure (≥ 500 mg/L) significantly delayed development and reduced adult lifespan (observed in females at 1,000 and males at 500–1,000 mg/L). Notably, trans-generational analysis revealed persistent developmental delays in the F1 generation, with offspring of 1,000 mg/L-exposed parents showing prolonged larval pupation time despite normal adult emergence timelines, indicative of partial developmental recovery. Mechanistically, high-dose PS disrupted endocrine signaling and altered expression of key developmental pathway regulators (EcR, InR, TOR, and E74B). These transcriptional changes were largely reversible in offspring, further supporting a similar gradually wash out recovery. While gut microbiota remained stable in exposed parents, offspring of 1,000 mg/L-exposed flies had significant microbiome alterations, highlighting clear trans-generational dysbiosis. This study provides new evidence that PS exposure during a sensitive developmental period in D. melanogaster can perturb development and gut microbial homeostasis across generations, offering insights relevant to understanding how early-life chemical exposures might influence conserved biological processes in higher organisms.
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Frontiers in Pharmacology
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2025
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IF 4.8
Fangfei Zhou,Qingge Lu,Lingyu Kong,Sitong Wang,Haixia Zhang,Meng Zhao,Yue Hu,Fanwu Wu,Chenxi Wu
Background: Cancer is a global public health crisis and the leading cause of death among middle-aged and older individuals, with its incidence increasingly shifting toward younger populations. Approximately 90% of the patients succumb to advanced metastasis, and effective treatments remain elusive. The specific molecular mechanisms underlying cancer cell invasion and migration remain poorly understood, hindering the development of effective targeted therapies. Therefore, inhibiting or reversing cancer cell invasion and migration may be crucial for reducing mortality. Our previous research revealed that the five drugs (FD), derived from Xuefu Zhuyu Decoction (XFZYD), play a significant role in inhibiting cell migration.Hypothesis/purpose: This study aims to explore the main drug components of FD and investigate the underlying mechanism in inhibiting cell migration.Methods: We used the Drosophila ptc>scrib-IR cell migration model to investigate the effects of FD. FD was disassembled and analyzed using an orthogonal design. Drug extracts were prepared and administered to Drosophila larvae. We assessed the effects of FD on cell migration, reactive oxygen species (ROS) levels, and gene expression.Results: In FD disassembled recipes and orthogonal test design, a significant difference was observed in the intervention with or without Glycyrrhiza uralensis Fisch. (GUF) in migrating cell number (P < 0.01), which emerged as a more potent inhibitor of FD from XFZYD in cell migration. High-performance liquid chromatography revealed that GUF and its extract contained effective medicinal components, namely glycyrrhizic acid, liquiritin, liquiritigenin, and glycyrrhetinic acid. Moreover, GUF at 4.0 mg/mL displayed strong inhibitory effect in migrating cell number and distance when compared with model, XFZYD or FD. Excessive ROS can activate the JAK/STAT signaling pathway and promote the EMT process. GUF inhibited ptc>scrib-IR-induced cell migration by reducing ROS levels, JAK/STAT signalling, and the transcription of upd2, upd3, hop and socs36E. Finally, GUF rescued the altered expressions of the epithelial-mesenchymal transition (EMT)-related proteins, including matrix metalloproteinase 1 (MMP1), β-integrin and E-cadherin, triggered by cell migration.Conclusion: Our findings demonstrate that GUF may serve as a promising candidate for targeting advanced metastatic tumors by suppressing ROS-mediated JAK/STAT signaling and EMT.
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Journal of Environmental Chemical Engineering
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2025
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IF 7.2
QiMeng He,Hui Jin,Jie Shen
Di (2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer, and many organisms encounter varying degrees of exposure in their natural environment. This study aimed to assess the impact of DEHP on Drosophila larvae by analyzing their behavior through kinematic parameters. We exposed the larvae to different concentrations of DEHP (0, 0.01%, 0.1%, 0.5%, 1% (v/v)), and their locomotor activity was observed, recorded, and the kinematic parameters were evaluated by machine learning method. The results indicated that the instantaneous velocity of larvae exposed to DEHP concentrations of 0.1%, 0.5%, and 1% significantly decreased, particularly in the 1% concentration group, where the instantaneous velocity dropped by 20.3%. Regarding instantaneous angular velocity, lower concentrations of DEHP resulted in an increase in instantaneous angular velocity; however, at a concentration of 1%, the instantaneous angular velocity significantly decreased by 22% compared to the control group. The results contribute to our understanding of the potential ecological and health risks posed to Insects and other organisms exposed to DEHP.
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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
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2025
·Vol. 122
·期 44
·e2513015122
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IF 9.1
Anqi Li,Dong Tong,Benjamin Ohlstein,Zheng Guo
Recent work suggests that asymmetric segregation of preexisting and newly synthesized canonical histone 3.1 (H3.1), but not variant histone 3.3A (H3.3A), plays an important role in the asymmetric outcome of Drosophila germline stem cell (GSC) and intestinal stem cell (ISC) divisions. However, this finding relied on the Gal4/UAS system and Flp-out technology to swap expression of exogenous fluorescent proteins—an approach that decouples H3.1 expression from the cell cycle and may be prone to artifacts. Here, by photoswitching photoconvertible H3-Dendra2 proteins expressed under their native controls in living flies, we find that preexisting and newly synthesized H3.1 and H3.3A are symmetrically segregated in dividing ISCs and somatic cyst stem cells (CySCs) in male testes. While H3.3A was found to be symmetrically segregated during GSC divisions, analysis of the H3.1 segregation in GSCs was complicated by our unexpected finding that H3.1-Dendra2 expressed under its normal control within the context of a histone transgene is not detectable in GSCs, owing to regulatory elements in the 3’UTR that mediate translational repression by the RNA-binding proteins Nanos and Pumilio. Nevertheless, when this repression is obviated by using a 3’UTR devoid of such elements, H3.1-Dendra2 is expressed in GSCs and symmetrically segregated without affecting lineage outcomes. We obtained similar results when the coding sequence of H3.3A was edited to encode H3.1 in the context of an H3.3A-Dendra2 knock-in allele. Our findings thus challenge the prevailing model of asymmetric histone segregation and reveal an unexpected molecular mechanism of posttranscriptional repression of H3.1 in GSCs.
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Antioxidants
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2022
·Vol. 12
·期 1
·59
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IF 7.675
La Yan,Xiaoxin Guo,Juanyu Zhou,Yuedan Zhu,Zehong Zhang,Haiyang Chen
Adult stem cells, a class of cells that possess self-renewal and differentiation capabilities, modulate tissue regeneration, repair, and homeostasis maintenance. These cells undergo functional degeneration during aging, resulting in decreased tissue regeneration ability and increased disease incidence. Thus, it is essential to provide effective therapeutic solutions to preventing the aging-related functional decline of stem cells. Quercetin (Que) is a popular natural polyphenolic flavonoid found in various plant species. It exhibits many beneficial effects against aging and aging-related diseases; however, its efficacy against adult stem cell aging remains largely unclear.Drosophilapossesses a mammalian-like intestinal system with a well-studied intestinal stem cell (ISC) lineage, making it an attractive model for adult stem cell research. Here, we show that Que supplementation could effectively prevent the hyperproliferation of ISCs, maintain intestinal homeostasis, and prolong the lifespan in agedDrosophila. In addition, we found that Que could accelerate recovery of the damaged gut and improve the tolerance ofDrosophilato stressful stimuli. Furthermore, results demonstrated that Que prevents the age-associated functional decline of ISCs via scavenging reactive oxygen species (ROS) and inhibiting the insulin signaling pathway. Overall, our findings suggest that Que plays a significant role in delaying adult stem cell aging.Keywords:intestinal stem cell;aging;quercetin;insulin signaling;ROS;Drosophila
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PEST MANAGEMENT SCIENCE
·
2023
·Vol. 79
·期 11
·4309-4318
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IF 4.1
Yaluan Bi,Xuan Zhang,Xuefei Chang,Jiaxin Li,Shan Xiao,Bo Zhang,Cong Dang,Linlin Sun,Hongwei Yao,Qi Fang,Fang Wang,Gongyin Ye
Background Behavior-based manipulation is an essential part of Drosophila integrated pest management (IPM). Effective compounds are useful for improving the efficiency of baits and the development of attract-and-kill or push–pull strategies to manage Drosophila populations. Here, we investigated the olfactory behavior of two Drosophila species, as well as their pupal parasitoid, to volatiles from bananas, for the identification of effective compounds to control fly populations. Results The results showed that overripe bananas were most attractive to both flies, with a higher attraction index (AI) in Drosophila melanogaster than in Drosophila suzukii . The profiles of volatile organic compounds (VOCs) from bananas with three different ripening stages were compared, and six VOCs were selected for behavioral tests. D. suzukii showed significantly different responses to isoamyl alcohol with D. melanogaster . The effects of ethyl butyrate and ethyl isovalerate on the two flies were dose-dependent, with lower concentrations acting as repellent and higher concentrations acting as attractant. Isoamyl acetate, isoamyl butyrate and isoamyl isovalerate (0.005–0.05% v/v) were attractive to both flies. The parasitoid wasp Pachycrepoidus vindemmiae was repelled by isoamyl alcohol and ethyl butyrate, and attracted by ethyl isovalerate, but showed no significant response to isoamyl acetate, isoamyl butyrate and isoamyl isovalerate at the tested concentration . Conclusion This study indicated that the behavioral response of Drosophila to the odor of chemical compounds is dose-dependent. Isoamyl alcohol and isoamyl acetate at certain concentrations could be used as repellents, whereas ethyl isovalerate as an attractant in control strategy design for D. suzukii. © 2023 Society of Chemical Industry.
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Insects
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2023
·Vol. 14
·期 12
·959
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IF 3
Jun-Hyoung Jeon,Seon-Ah Jeong,Doo-Sang Park,Hong-Hyun Park,Sang-Woon Shin,Hyun-Woo Oh
Simple SummarySeveral plant species have diterpene compounds with juvenile hormone (JH) disruptor activity in insects. Demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC) are two curcuminoid components of turmeric, interfering with the JH-mediated formation of the JH receptor complex. In vitro, DMC and BDMC also inhibit larval and pupal development inDrosophila melanogaster. The results suggest that both DMC and BDMC are JH disruptors that affect insect growth and development by regulating JH-mediated gene expression.AbstractJuvenile hormones (JHs) play a central role in insect development, reproduction, and various physiological functions. Curcuminoids generally exhibit a wide range of biological activities, such as antioxidant, anti-inflammatory, antibacterial, and insecticidal, and they exhibit insect growth inhibitory effects. However, research on insecticidal properties of curcuminoids has been limited. Moreover, to the best of our knowledge, studies on JHs of insects and curcuminoids are lacking. Therefore, this study aimed to identify the substances that act as JH disruptors (JHDs) from edible plants. Demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC), two curcuminoids from the turmeric plantCurcuma longaL. inhibited the formation of a methoprene-tolerant (Met)–Taiman (Tai) heterodimer complex inDrosophila melanogaster, as shown through in vitro yeast two-hybrid assays. An artificial diet containing 1% (w/v) DMC or BDMC significantly reduced the number ofD. melanogasterlarvae in a concentration-dependent manner; larval development was disrupted, preventing the progression of larvae to pupal stages, resulting in an absence of adults. Building on the results obtained in this study on curcuminoids, researchers can use our study as a reference to develop eco-friendly pesticides.Keywords:juvenile hormone;methoprene-tolerant;diterpene;juvenile hormone disruptor;curcuminoids;demethoxycurcumin;bisdemethoxycurcumin
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Cell and Bioscience
·
2023
·Vol. 13
·期 1
·1-20
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IF 7.5
Mi Kai,Li Yiqing,Yang Yuhang,Secombe Julie,Liu Xingyin
Background Drosophila melanogaster is excellent animal model for understanding the molecular basis of human neurological and motor disorders. The experimental conditions and chamber design varied between studies. Moreover, most previously established paradigms focus on fly trace detection algorithm development. A comprehensive understanding on how fly behaves in the chamber is still lacking. Results In this report, we established 74 unique behavior metrics quantifying spatiotemporal characteristics of adult fly locomotion and social behaviors, of which 49 were newly proposed. By the aiding of the developed analysis pipeline, Drosophila video tracking (DVT), we identified siginificantly different patterns of fly behavior confronted with different chamber height, fly density, illumination and experimental time. Meanwhile, three fly strains which are widely used as control lines, Canton-S(CS), w 1118 and Oregon-R (OR), were found to exhibit distinct motion explosiveness and exercise endurance. Conclusions We believe the proposed behavior metrics set and pipeline should help identify subtle spatial and temporal differences of drosophila behavior confronted with different environmental factors or gene variants.
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