Transcription factors of the bHLH-PAS family play vital roles in animal development, physiology, and disease. Two members of the family require binding of low-molecular weight ligands for their activity: the vertebrate aryl hydrocarbon receptor (AHR) and the insect juvenile hormone receptor (JHR). In the fly Drosophila melanogaster, the paralogous proteins GCE and MET constitute the ligand-binding component of JHR complexes. Whilst GCE/MET and AHR are phylogenetically heterologous, their mode of action is similar. JHR is targeted by several synthetic agonists that serve as insecticides disrupting the insect endocrine system. AHR is an important regulator of human endocrine homeostasis, and it responds to environmental pollutants and endocrine disruptors. Whether AHR signaling is affected by compounds that can activate JHR has not been reported. To address this question, we screened a chemical library of 50,000 compounds to identify 93 novel JHR agonists in a reporter system based on Drosophila cells. Of these compounds, 26% modulated AHR signaling in an analogous reporter assay in a human cell line, indicating a significant overlap in the agonist repertoires of the two receptors. To explore the structural features of agonist-dependent activation of JHR and AHR, we compared the ligand-binding cavities and their interactions with selective and common ligands of AHR and GCE. Molecular dynamics modeling revealed ligand-specific as well as conserved side chains within the respective cavities. Significance of predicted interactions was supported through site-directed mutagenesis. The results have indicated that synthetic insect juvenile hormone agonists might interfere with AHR signaling in human cells.

• Klíčová slova
• aryl hydrocarbon receptor, endocrine disruptors, high-throughput screening, juvenile hormone receptor, ligand binding domain,
• MeSH
• buněčné linie MeSH
• Drosophila melanogaster * metabolismus   genetika   účinky léků   MeSH
• juvenilní hormony metabolismus   farmakologie   MeSH
• lidé MeSH
• ligandy MeSH
• proteiny Drosophily * metabolismus   genetika   chemie   MeSH
• receptory aromatických uhlovodíků * agonisté   metabolismus   genetika   MeSH
• signální transdukce účinky léků   MeSH
• transkripční faktory bHLH * metabolismus   genetika   agonisté   MeSH
• transkripční faktory metabolismus   genetika   MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• AHR protein, human MeSH Prohlížeč
• gce protein, Drosophila MeSH Prohlížeč
• juvenilní hormony MeSH
• ligandy MeSH
• MET protein, Drosophila MeSH Prohlížeč
• proteiny Drosophily * MeSH
• receptory aromatických uhlovodíků * MeSH
• transkripční faktory bHLH * MeSH
• transkripční faktory MeSH

Repression of msl-2 mRNA translation is essential for viability of Drosophila melanogaster females to prevent hypertranscription of both X chromosomes. This translational control event is coordinated by the female-specific protein Sex-lethal (Sxl) which recruits the RNA binding proteins Unr and Hrp48 to the 3' untranslated region (UTR) of the msl-2 transcript and represses translation initiation. The mechanism exerted by Hrp48 during translation repression and its interaction with msl-2 are not well understood. Here we investigate the RNA binding specificity and affinity of the tandem RNA recognition motifs of Hrp48. Using NMR spectroscopy, molecular dynamics simulations and isothermal titration calorimetry, we identified the exact region of msl-2 3' UTR recognized by Hrp48. Additional biophysical experiments and translation assays give further insights into complex formation of Hrp48, Unr, Sxl and RNA. Our results show that Hrp48 binds independent of Sxl and Unr downstream of the E and F binding sites of Sxl and Unr to msl-2.

• Klíčová slova
• Dosage compensation, Hrp48, RNA binding protein, RNA recognition motif, Translation regulation,
• MeSH
• 3' nepřekládaná oblast * MeSH
• DNA vazebné proteiny MeSH
• Drosophila melanogaster * metabolismus   genetika   MeSH
• heterogenní jaderné ribonukleoproteiny MeSH
• messenger RNA metabolismus   genetika   chemie   MeSH
• proteiny Drosophily * metabolismus   chemie   genetika   MeSH
• proteiny vázající RNA * metabolismus   chemie   genetika   MeSH
• proteosyntéza MeSH
• simulace molekulární dynamiky MeSH
• transkripční faktory metabolismus   chemie   genetika   MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 3' nepřekládaná oblast * MeSH
• DNA vazebné proteiny MeSH
• heterogenní jaderné ribonukleoproteiny MeSH
• Hrb27C protein, Drosophila MeSH Prohlížeč
• messenger RNA MeSH
• msl-2 protein, Drosophila MeSH Prohlížeč
• proteiny Drosophily * MeSH
• proteiny vázající RNA * MeSH
• transkripční faktory MeSH

The widely used insecticide chlorpyrifos (CP) is known to inhibit acetylcholinesterase (AChE) activity attributed to result in various neurological disorders and acetylcholine-dependent organ functions including heart, skeletal muscle, lung, gastrointestinal tract, and central nervous systems. Enzyme reactivators, such as oximes, are known to restore AChE activity and mitigate adverse effects. The identification of compounds that reactivate AChE constitute agents with important therapeutic beneficial effects in cases of pesticide poisoning. However, the screening of novel drugs using traditional models may raise ethical concerns. This study aimed to investigate the potential of Drosophila melanogaster as a model organism for screening AChE reactivators, with a focus on organophosphate poisoning. The efficacy of several oximes, including pralidoxime, trimedoxime, obidoxime, methoxime, HI-6, K027, and K048, against CP-induced AChE activity inhibition in D. melanogaster was determined in silico, in vitro, and in vivo experiments. Molecular docking studies indicated a strong interaction between studied oximes and the active-site gorge of AChE. Data showed that selected oximes (100 μM) are effective in the reactivation of AChE inhibited by CP (10 μM) in vitro. Finally, in vivo investigations demonstrated that selected oximes, pralidoxime and K048 (1.5 ppm), reversed the locomotor deficits, inhibition of AChE activity as well as lowered the mortality rates induced by CP (0.75 ppm). Our findings contribute to utilization of D. melanogaster as a robust model for determination of actions of identified new AChE inhibitory agents with more effective therapeutic properties that those currently in use in the clinical practice in treatment of AChE associated disorders.

• Klíčová slova
• Drosophila melanogaster, Insecticides, acetylcholinesterase inhibition, locomotor, molecular docking, organophosphates, oximes, risk assessment, screening,
• MeSH
• acetylcholinesterasa * metabolismus   MeSH
• cholinesterasové inhibitory toxicita   MeSH
• Drosophila melanogaster * účinky léků   enzymologie   MeSH
• dursban * toxicita   MeSH
• insekticidy toxicita   MeSH
• modely u zvířat MeSH
• oximy * farmakologie   MeSH
• reaktivátory cholinesterázy * farmakologie   MeSH
• simulace molekulového dockingu * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acetylcholinesterasa * MeSH
• cholinesterasové inhibitory MeSH
• dursban * MeSH
• insekticidy MeSH
• oximy * MeSH
• reaktivátory cholinesterázy * MeSH

The minute wasp Habrobracon hebetor venom (HH venom) is a potent cocktail of toxins that paralyzes the victim's muscles and suppresses humoral and cellular immunity. This study examined the effect of HH venom on specific biochemical, physiological, and ultrastructural characteristics of the thoracic and nervous (CNS) tissues of Drosophila melanogaster under in vitro conditions. Venom treatment modulated the activities of superoxide dismutase (SOD) and catalase (CAT), endogenous Drome-AKH level, and affected the relative viability of the cells. Additionally, it reduced the expression of genes related to the immune system in the CNS, including Keap1, Relish, Nox, Eiger, Gadd45, and Domeless, as well as in the thoracic muscles, except for Nox. Besides, venom treatment led to deteriorative changes in the ultrastructure of muscle cells, particularly affecting the mitochondria. When venom and Drosophila melanogaster-adipokinetic hormone (Drome-AKH) were applied together, the effects of the venom alone were often modulated. The harmful effect of the venom on SOD activity was relatively reduced and the activity returned to a level similar to that of the control. In the CNS, the simultaneous application of venom and hormones abolished the suppression of previously reported immune-related genes (except for Gadd45), whereas in the muscles, this was only true for Eiger. Additionally, Drome-AKH restored cell structure to a level comparable to that of the control and lessened the harmful effects of HH venom on muscle mitochondria. These findings suggest a general body response of D. melanogaster to HH venom and a partial defensive role of Drome-AKH in this process.

• Klíčová slova
• Adipokinetic hormone, Habrobracon venom, Immune responsive genes, Muscle structure, Oxidative stress,
• MeSH
• centrální nervový systém účinky léků   MeSH
• Drosophila melanogaster * účinky léků   MeSH
• hmyzí hormony * metabolismus   MeSH
• katalasa metabolismus   genetika   MeSH
• kyselina pyrrolidonkarboxylová * analogy a deriváty   MeSH
• oligopeptidy * farmakologie   toxicita   MeSH
• proteiny Drosophily * metabolismus   genetika   MeSH
• sršňovití * účinky léků   MeSH
• superoxiddismutasa metabolismus   MeSH
• vosí jedy * toxicita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adipokinetic hormone MeSH Prohlížeč
• hmyzí hormony * MeSH
• katalasa MeSH
• kyselina pyrrolidonkarboxylová * MeSH
• oligopeptidy * MeSH
• proteiny Drosophily * MeSH
• superoxiddismutasa MeSH
• vosí jedy * MeSH

In multicellular organisms, the indole melatonin synthesized by aralkylamine N-acetyltransferase (AANATI) serves as an antioxidant. To test this, sex-mixed 3-day-old mated fly adults bw1 and AANAT1 homozygous recessive loss-of-function mutant (bw AANAT1lo) of Drosophila melanogaster were fed by a standard diet or by one containing paraquat (PQ, 1,1'-dimethyl-4,4'-bipyridilium dichloride hydrate) at a final concentration of 15.5 mM. Experiment lasted 8 h and began at 11 a.m. In bw1 flies the paraquat treatment resulted in a significant (evaluated by Student's t-tests) decrease of the superoxide dismutase (SOD) activity and an increase the catalase (CAT) and glutathione S-transferase (GST) activities. Meanwhile, in these flies, total Antioxidative capacity (TAC) was significantly curbed by the paraquat presence. Importantly, these changes were not observed in the AANAT1-mutants. Thus, melatonin seems to play an important defence role against the oxidative stress elicited by paraquat.

• Klíčová slova
• arylalkylamine‐N‐acetyltransferase, drosophila, mutant, oxidative stress, paraquat, stress response,
• MeSH
• acetyltransferasy metabolismus   genetika   MeSH
• antioxidancia * metabolismus   MeSH
• Drosophila melanogaster * účinky léků   enzymologie   MeSH
• glutathiontransferasa * metabolismus   genetika   MeSH
• katalasa metabolismus   genetika   MeSH
• melatonin farmakologie   metabolismus   MeSH
• oxidační stres * účinky léků   MeSH
• paraquat * farmakologie   MeSH
• proteiny Drosophily metabolismus   genetika   MeSH
• superoxiddismutasa metabolismus   genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetyltransferasy MeSH
• antioxidancia * MeSH
• glutathiontransferasa * MeSH
• katalasa MeSH
• melatonin MeSH
• paraquat * MeSH
• proteiny Drosophily MeSH
• superoxiddismutasa MeSH

Insect prophenoloxidases (PPOs) are important immunity proteins for defending against the invading pathogens and parasites. As a Type-Ⅲ copper-containing proteins, unlike Homo sapiens tyrosinases, the insect PPOs and most bacterial tyrosinases contain no signal peptides for unknown reason, however they can still be released. To this end, we fused different signal peptides to Drosophila melanogaster PPOs for in vitro and in vivo expression, respectively. We demonstrate that an artificial signal peptide can help PPO secretion in vitro. The secreted PPO appeared larger than wild-type PPO on molecular weight sizes due to glycosylation when expressed in S2 cells. Two asparagine residues for potential glycosylation in PPO1 were identified when a signal peptide was fused. After purification, the glycosylated PPO1 lost zymogen activity. When PPO1 containing a signal peptide was over-expressed in Drosophila larvae, the glycosylation and secretion of PPO1 was detected in vivo. Unlike insect PPO, human tyrosinase needs a signal peptide for protein expression and maintaining enzyme activity. An artificial signal peptide fused to bacterial tyrosinase had no influence on the protein expression and enzyme activity. These Type-Ⅲ copper-containing proteins from different organisms may evolve to perform their specific functions. Intriguingly, our study revealed that the addition of calcium inhibits PPO secretion from the transiently cultured larval hindguts in vitro, indicating that the calcium concentration may regulate PPO secretion. Taken together, insect PPOs can maintain enzyme activities without any signal peptide.

• Klíčová slova
• Enzyme activity, Glycosylation, Insect, Prophenoloxidase, Signal peptide,
• MeSH
• buněčné linie MeSH
• Drosophila melanogaster * imunologie   metabolismus   MeSH
• glykosylace MeSH
• hmyzí proteiny metabolismus   genetika   MeSH
• katecholoxidasa * metabolismus   MeSH
• larva metabolismus   MeSH
• lidé MeSH
• prekurzory enzymů * metabolismus   MeSH
• proteinové prekurzory metabolismus   MeSH
• proteiny - lokalizační signály * MeSH
• proteiny Drosophily metabolismus   genetika   MeSH
• tyrosinasa metabolismus   MeSH
• vápník metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hmyzí proteiny MeSH
• katecholoxidasa * MeSH
• prekurzory enzymů * MeSH
• pro-phenoloxidase MeSH Prohlížeč
• proteinové prekurzory MeSH
• proteiny - lokalizační signály * MeSH
• proteiny Drosophily MeSH
• tyrosinasa MeSH
• vápník MeSH

Dietary restriction (DR) slows aging in many animals, while in some cases, the sensory signals from diet alone are sufficient to retard or accelerate lifespan. The digestive tract is a candidate location to sense nutrients, where neuropeptides secreted by enteroendocrine cells (EEC) produce systemic signals in response to food. Here, we measure how Drosophila neuropeptide F (NPF) is secreted into adult circulation by EEC and find that specific EEC differentially respond to dietary sugar and yeast. Female lifespan is increased when gut NPF is genetically depleted, and this manipulation is sufficient to blunt the longevity benefit conferred by DR. Depletion of NPF receptors at insulin-producing neurons of the brain also increases female lifespan, consistent with observations where loss of gut NPF decreases neuronal insulin secretion. The longevity conferred by repressing gut NPF and brain NPF receptors is reversed by treating adults with a juvenile hormone (JH) analog. JH is produced by the adult corpora allata, and inhibition of the insulin receptor at this tissue decreases JH titer and extends lifespan in both males and females, while this longevity is restored to wild type by treating adults with a JH analog. Overall, EEC of the gut modulate Drosophila aging through interorgan communication mediated by a gut-brain-corpora allata axis, and insulin produced in the brain impacts lifespan through its control of JH titer. These data suggest that we consider how human incretins and their analogs, which are used to treat obesity and diabetes, may impact aging.

• Klíčová slova
• aging, incretin, insulin, interorgan communication, juvenile hormone,
• MeSH
• dlouhověkost fyziologie   MeSH
• Drosophila melanogaster metabolismus   MeSH
• enteroendokrinní buňky metabolismus   MeSH
• inzulin * metabolismus   MeSH
• juvenilní hormony * metabolismus   MeSH
• mozek metabolismus   MeSH
• neurony metabolismus   MeSH
• neuropeptidy * metabolismus   MeSH
• osa mozek-střevo * fyziologie   MeSH
• proteiny Drosophily * metabolismus   genetika   MeSH
• stárnutí metabolismus   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• inzulin * MeSH
• juvenilní hormony * MeSH
• neuropeptide F, Drosophila MeSH Prohlížeč
• neuropeptidy * MeSH
• proteiny Drosophily * MeSH

Pheromones convey rich ethological information and guide insects' search behavior. Insects navigating in turbulent environments are tasked with the challenge of coding the temporal structure of an odor plume, obliging recognition of the onset and offset of whiffs of odor. The coding mechanisms that shape odor offset recognition remain elusive. We designed a device to deliver sharp pheromone pulses and simultaneously measured the response dynamics from pheromone-tuned olfactory receptor neurons (ORNs) in male moths and Drosophila. We show that concentration-invariant stimulus duration encoding is implemented in moth ORNs by spike frequency adaptation at two time scales. A linear-nonlinear model fully captures the underlying neural computations and offers an insight into their biophysical mechanisms. Drosophila use pheromone cis-vaccenyl acetate (cVA) only for very short distance communication and are not faced with the need to encode the statistics of the cVA plume. Their cVA-sensitive ORNs are indeed unable to encode odor-off events. Expression of moth pheromone receptors in Drosophila cVA-sensitive ORNs indicates that stimulus-offset coding is receptor independent. In moth ORNs, stimulus-offset coding breaks down for short ( < 200 ms) whiffs. This physiological constraint matches the behavioral latency of switching from the upwind surge to crosswind cast flight upon losing contact with the pheromone.

• MeSH
• acetáty MeSH
• čich fyziologie   MeSH
• čichové buňky * fyziologie   MeSH
• čichové dráhy fyziologie   MeSH
• Drosophila melanogaster fyziologie   MeSH
• Drosophila fyziologie   MeSH
• feromony * MeSH
• kyseliny olejové MeSH
• můry * fyziologie   MeSH
• odoranty analýza   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetáty MeSH
• cis-vaccenyl acetate MeSH Prohlížeč
• feromony * MeSH
• kyseliny olejové MeSH

One of the major functions of programmed cell death (apoptosis) is the removal of cells that suffered oncogenic mutations, thereby preventing cancerous transformation. By making use of a Double-Headed-EP (DEP) transposon, a P element derivative made in our laboratory, we made an insertional mutagenesis screen in Drosophila melanogaster to identify genes that, when overexpressed, suppress the p53-activated apoptosis. The DEP element has Gal4-activatable, outward-directed UAS promoters at both ends, which can be deleted separately in vivo. In the DEP insertion mutants, we used the GMR-Gal4 driver to induce transcription from both UAS promoters and tested the suppression effect on the apoptotic rough eye phenotype generated by an activated UAS-p53 transgene. By DEP insertions, 7 genes were identified, which suppressed the p53-induced apoptosis. In 4 mutants, the suppression effect resulted from single genes activated by 1 UAS promoter (Pka-R2, Rga, crol, and Spt5). In the other 3 (Orct2, Polr2M, and stg), deleting either UAS promoter eliminated the suppression effect. In qPCR experiments, we found that the genes in the vicinity of the DEP insertion also showed an elevated expression level. This suggested an additive effect of the nearby genes on suppressing apoptosis. In the eukaryotic genomes, there are coexpressed gene clusters. Three of the DEP insertion mutants are included, and 2 are in close vicinity of separate coexpressed gene clusters. This raises the possibility that the activity of some of the genes in these clusters may help the suppression of the apoptotic cell death.

• Klíčová slova
• Drosophila, activating insertional mutagenesis, apoptosis, p53, suppression,
• MeSH
• apoptóza * MeSH
• dominantní geny MeSH
• Drosophila melanogaster * genetika   MeSH
• fenotyp MeSH
• inzerční mutageneze * metody   MeSH
• nádorový supresorový protein p53 * genetika   metabolismus   MeSH
• promotorové oblasti (genetika) MeSH
• proteiny Drosophily * genetika   metabolismus   MeSH
• supresorové geny MeSH
• transpozibilní elementy DNA MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• nádorový supresorový protein p53 * MeSH
• p53 protein, Drosophila MeSH Prohlížeč
• proteiny Drosophily * MeSH
• transpozibilní elementy DNA MeSH

Adjustment of the cellular metabolism of pro-inflammatory macrophages is essential for their bactericidal function; however, it underlies the development of many human diseases if induced chronically. Therefore, intervention of macrophage metabolic polarisation has been recognised as a potent strategy for their treatment. Although many small-molecule inhibitors affecting macrophage metabolism have been identified, their in vivo administration requires a tool for macrophage-specific delivery to limit their potential side effects. Here, we establish Drosophila melanogaster as a simple experimental model for in vivo testing of macrophage-specific delivery tools. We found that yeast-derived glucan particles (GPs) are suitable for macrophage-specific delivery of small-molecule inhibitors. Systemic administration of GPs loaded with atorvastatin, the inhibitor of hydroxy-methyl-glutaryl-CoA reductase (Hmgcr), leads to intervention of mevalonate pathway specifically in macrophages, without affecting HMGCR activity in other tissues. Using this tool, we demonstrate that mevalonate pathway is essential for macrophage pro-inflammatory polarisation and individual's survival of infection.

• Klíčová slova
• atorvastatin, glucan particles, macrophage‐specific delivery, mevalonate pathway,
• MeSH
• atorvastatin * farmakologie   aplikace a dávkování   MeSH
• Drosophila melanogaster * MeSH
• glukany farmakologie   MeSH
• hydroxymethylglutaryl-CoA-reduktasy metabolismus   MeSH
• kyselina mevalonová * metabolismus   MeSH
• makrofágy * metabolismus   účinky léků   MeSH
• statiny farmakologie   aplikace a dávkování   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• atorvastatin * MeSH
• glukany MeSH
• hydroxymethylglutaryl-CoA-reduktasy MeSH
• kyselina mevalonová * MeSH
• statiny MeSH