Adult Fly Activity Research Articles

Effect of Malathion on Catalase Enzyme and Acetylcholinesterase Activity in Adult Flies and 3rd Instar Larvae of Drosophila melanogaster

Effect of Malathion on Catalase Enzyme and Acetylcholinesterase Activity in Adult Flies and 3rd Instar Larvae of Drosophila melanogaster

Optogenetic cleavage of the Miro GTPase reveals the direct consequences of real-time loss of function in Drosophila.

Miro GTPases control mitochondrial morphology, calcium homeostasis, and regulate mitochondrial distribution by mediating their attachment to the kinesin and dynein motor complex. It is not clear, however, how Miro proteins spatially and temporally integrate their function as acute disruption of protein function has not been performed. To address this issue, we have developed an optogenetic loss of function "Split-Miro" allele for precise control of Miro-dependent mitochondrial functions in Drosophila. Rapid optogenetic cleavage of Split-Miro leads to a striking rearrangement of the mitochondrial network, which is mediated by mitochondrial interaction with the microtubules. Unexpectedly, this treatment did not impact the ability of mitochondria to buffer calcium or their association with the endoplasmic reticulum. While Split-Miro overexpression is sufficient to augment mitochondrial motility, sustained photocleavage shows that Split-Miro is surprisingly dispensable to maintain elevated mitochondrial processivity. In adult fly neurons in vivo, Split-Miro photocleavage affects both mitochondrial trafficking and neuronal activity. Furthermore, functional replacement of endogenous Miro with Split-Miro identifies its essential role in the regulation of locomotor activity in adult flies, demonstrating the feasibility of tuning animal behaviour by real-time loss of protein function.

Artificial diet alters activity and rest patterns in the olive fruit fly.

Olive fruit flies, Bactrocera oleae (Diptera: Tephritidae) reared in the laboratory on an artificial diet are essential for the genetic control techniques against this pest. However, the colony's laboratory adaptation can affect the quality of the reared flies. We used the Locomotor Activity Monitor to track the activity and rest patterns of adult olive fruit flies reared as immatures in olives (F2-F3 generation) and in artificial diet (>300 generations). Counts of beam breaks caused by the adult fly activity were used as an estimation of its locomotor activity levels during the light and dark period. Bouts of inactivity with duration longer than five minutes were considered a rest episode. Locomotor activity and rest parameters were found to be dependent on sex, mating status and rearing history. In virgin flies reared on olives, males were more active than females and increased their locomotor activity towards the end of the light period. Mating decreased the locomotor activity levels of males, but not of female olive-reared flies. Laboratory flies reared on artificial diet had lower locomotor activity levels during the light period and more rest episodes of shorter duration during the dark period compared to flies reared on olives. We describe the diurnal locomotor activity patterns of B. oleae adults reared on olive fruit and on artificial diet. We discuss how locomotor activity and rest pattern differences may affect the laboratory flies' ability to compete with wild males in the field.

The CRTC-CREB axis functions as a transcriptional sensor to protect against proteotoxic stress in Drosophila

cAMP Responsible Element Binding Protein (CREB) is an evolutionarily conserved transcriptional factor that regulates cell growth, synaptic plasticity and so on. In this study, we unexpectedly found proteasome inhibitors, such as MLN2238, robustly increase CREB activity in adult flies through a large-scale compound screening. Mechanistically, reactive oxidative species (ROS) generated by proteasome inhibition are required and sufficient to promote CREB activity through c-Jun N-terminal kinase (JNK). In 293 T cells, JNK activation by MLN2238 is also required for increase of CREB phosphorylation at Ser133. Meanwhile, transcriptome analysis in fly intestine identified a group of genes involved in redox and proteostatic regulation are augmented by overexpressing CRTC (CREB-regulated transcriptional coactivator). Intriguingly, CRTC overexpression in muscles robustly restores protein folding and proteasomal activity in a fly Huntington’s disease (HD) model, and ameliorates HD related pathogenesis, such as protein aggregates, motility, and lifespan. Moreover, CREB activity increases during aging, and further enhances its activity can suppress protein aggregates in aged muscles. Together, our results identified CRTC/CREB downstream ROS/JNK signaling as a conserved sensor to tackle oxidative and proteotoxic stresses. Boosting CRTC/CREB activity is a potential therapeutic strategy to treat aging related protein aggregation diseases.

Review of Methods to Monitor House Fly (Musca domestica) Abundance and Activity.

The house fly is a ubiquitous pest commonly associated with animal facilities and urban waste. When present in large numbers, house flies can negatively impact humans and animals through nuisance and the transmission of pathogens. Since the development of fly traps and sticky papers to capture flies in the late 1800s, these and other methods have been used as a means to monitor change in house fly density or fly activity over time. Methods include substrate sampling to record density of immature flies, visual observations of adult fly activity, instantaneous counts of landing or resting flies, accumulation of adult flies on/in traps, or accumulation of fly fecal and regurgitation spots deposited by flies onto white cards. These methods do not estimate true house fly density, but rather provide an index of house fly activity that is related to both fly density and the frequency of individual fly behavior (e.g., frequency of flight, landing events) and which is likely more predictive of negative impacts such as nuisance and pathogen transmission. Routine monitoring of house fly activity is a critical component of a house fly management program. Fly activity should be held to a level below a predetermined activity threshold ('action threshold') above which negative impacts are anticipated to occur. This article is a review of methods utilized for monitoring house fly (Diptera: Muscidae) activity.

Health inequality and COVID-19: the culmination of two centuries of social murder.

<h3>ABSTRACT</h3> Miro GTPases control mitochondrial morphology, calcium homeostasis and regulate mitochondrial distribution by mediating their attachment to the kinesin and dynein motor complex. It is not clear, however, how Miro proteins spatially and temporally integrate their function as acute disruption of protein function has not been performed. To address this issue, we have developed an optogenetic loss of function ‘Split-Miro’ allele for precise control of Miro-dependent mitochondrial functions in <i>Drosophila</i>. Rapid optogenetic cleavage of Split-Miro leads to a striking rearrangement of the mitochondrial network, which is mediated by mitochondrial interaction with the microtubules. Unexpectedly, this treatment did not impact the ability of mitochondria to buffer calcium. While Split-Miro overexpression is sufficient to augment mitochondrial motility, sustained photocleavage shows Split-Miro is surprisingly dispensable to maintain elevated mitochondrial processivity. Furthermore, functional replacement of endogenous Miro with Split-Miro identifies its essential role in the regulation of locomotor activity in adult flies, demonstrating the feasibility of tuning animal behaviour by real-time loss of protein function.

The Phosphatase CSW Controls Life Span by Insulin Signaling and Metabolism Throughout Adult Life in Drosophila.

Noonan syndrome and related disorders are caused by mutations in genes encoding for proteins of the RAS-ERK1/2 signaling pathway, which affect development by enhanced ERK1/2 activity. However, the mutations’ effects throughout adult life are unclear. In this study, we identify that the protein most commonly affected in Noonan syndrome, the phosphatase SHP2, known in Drosophila as corkscrew (CSW), controls life span, triglyceride levels, and metabolism without affecting ERK signaling pathway. We found that CSW loss-of-function mutations extended life span by interacting with components of the insulin signaling pathway and impairing AKT activity in adult flies. By expressing csw-RNAi in different organs, we determined that CSW extended life span by acting in organs that regulate energy availability, including gut, fat body and neurons. In contrast to that in control animals, loss of CSW leads to reduced homeostasis in metabolic rate during activity. Clinically relevant gain-of-function csw allele reduced life span, when expressed in fat body, but not in other tissues. However, overexpression of a wild-type allele did not affect life span, showing a specific effect of the gain-of-function allele independently of a gene dosage effect. We concluded that CSW normally regulates life span and that mutations in SHP2 are expected to have critical effects throughout life by insulin-dependent mechanisms in addition to the well-known RAS-ERK1/2-dependent developmental alterations.

Novel genetic link between the ATP-binding cassette subfamily A gene and hippo gene in Drosophila

The pan-neuron-specific knockdown of dABCA, a Drosophila homologue of the human ATP binding cassette subfamily A member 13 gene, increases social space without affecting climbing ability and induces the early onset of evening activity in adult flies followed by relatively high activity throughout the day. Satellite bouton numbers in the presynaptic terminals of motor neurons are increased in dABCA knockdown flies. In the present study, we further characterized pan-neuron-specific dABCA knockdown flies and found that active zones in the presynaptic terminals of motor neurons increased, whereas learning abilities decreased in larvae. Genetic crossing experiments revealed that the hippo mutation enhanced the hyperactivity phenotype of adults, but suppressed the increased satellite bouton phenotype induced by the dABCA knockdown. Drosophila ABCA is predicted to transport lipid molecules and impair the asymmetric distribution of phospholipids across the plasma membrane, and these local changes are considered to be important for various cellular functions. The disruption of lipid homeostasis in central and peripheral nervous systems by the dABCA knockdown may affect the Hippo-related signaling pathway in order to induce the observed phenotypes.

Wild bacterial probiotics fed to larvae of mass-reared Queensland fruit fly [Bactrocera tryoni (Froggatt)] do not impact long-term survival, mate selection, or locomotor activity.

Queensland fruit fly [Bactrocera tryoni (Froggatt), Diptera, Tephritidae] is the most devastating insect pest impacting Australian horticulture. The Sterile Insect Technique (SIT) is an important component of tephritid pest management programs. However, mass-rearing and irradiation (to render insects sterile) may reduce the fitness and performance of the insect, including the ability of sterile males to successfully compete for wild females. Manipulation of the gut microbiome, including the supplementation with bacterial probiotics shows promise for enhancing the quality of mass-reared sterile flies, however there are fewer published studies targeting the larval stage. In this study, we supplemented the larval stage of mass-reared B. tryoni with bacterial probiotics. We tested several individual bacteria that had been previously isolated and characterized from the gut of wild B. tryoni larvae including Asaia sp., Enterobacter sp., Lactobacillus sp., Leuconostoc sp. We also tested a consortium of all four of these bacterial isolates. The fitness parameters tested included adult survival in field cages, laboratory mate selection of bacteria supplemented males by bacteria nonsupplemented females, and laboratory locomotor activity of adult flies. None of the bacterial probiotic treatments in the current study was significantly different to the control for field survival, mate selection or locomotor activity of adult B. tryoni, which agree with some of the other studies regarding bacterial probiotics fed to the larval stage of tephritids. Future work is needed to determine if feeding the same, and/or other probiotics to adults, as opposed to larvae can positively impact survival, mating performance, mating competitiveness and locomotor activity of B. tryoni. The bacterial group(s) and function of bacterial species that increase fitness and competitiveness is also of interest to tephritid mass-rearing programs.

Novel Drosophila model for psychiatric disorders including autism spectrum disorder by targeting of ATP-binding cassette protein A

Autism spectrum disorder (ASD) is characterized by persistent deficits in social communication and social interactions, as well as restricted, stereotyped patterns of behavior and interests. In addition, alterations in circadian sleep-wake rhythm are common in young children with ASD. Mutations in ATP binding cassette subfamily A member 13 (ABCA13) have been recently identified in a monkey that displays behavior associated with ASD. ABCA13, a member of the ABCA family of proteins, is predicted to transport lipid molecules and is expressed in the human trachea, testis, bone marrow, hippocampus, cortex, and other tissues. However, its physiological function remains unknown. Drosophila CG1718 shows high homology to human ABCA genes including ABCA13 and is thus designated as Drosophila ABCA (dABCA). To elucidate the physiological role of dABCA, we specifically knocked down dABCA in all neurons of flies and investigated their phenotypes. The pan-neuron-specific knockdown of dABCA resulted in increased social space with the closest neighbor in adult male flies but exerted no effect on their climbing ability, indicating that the increase in social space is not due to a defect in their climbing ability. An activity assay with adult male flies revealed that knockdown of dABCA in all neurons induces early onset of evening activity in adult flies followed by relatively high activity during morning peaks, evening peaks, and midday siesta. These phenotypes are similar to defects observed in human ASD patients, suggesting that the established dABCA knockdown flies are a promising model for ASD. In addition, an increase in satellite boutons in presynaptic terminals of motor neurons was observed in dABCA knockdown third instar larvae, suggesting that dABCA regulates the formation and/or maintenance of presynaptic terminals of motor neurons.

Field evaluation of a semi-automatic funnel trap targeted the medically important non-biting flies

Bait-trapping is a useful approach for monitoring fly population dynamics, and it is an effective tool for physical control of pest species. The aim of this study was to test a newly developed semi-automatic funnel fly trap with some modifications of the former prototype fly trap to study medically important fly population density. The efficacy of the semi-automatic funnel trap was assessed by field sampling during July 2013-June 2014 using 1-day tainted beef offal as bait. The modified semi-automatic funnel traps were able to capture a total of 151,141 adult flies, belonging to the families: Calliphoridae (n=147,248; 97.4%), Muscidae (n=3,124; 2.1%) and Sarcophagidae (n=769; 0.5%), which are the medically important fly species. Among the total of 35 species collected, Chrysomya megacephala (Diptera: Calliphoridae) (n=88,273; 59.95%), Musca domestica (Diptera: Muscidae) (n=1,324; 42.38%) and Boettcherisca peregrina (Diptera: Sarcophagidae) (n=68; 33.01%) were the predominant species of each family. High number of flies was captured in forest area, representing 42.47% (n=64,197) of total specimens. Female flies were trapped more than male with total sex ratio of 0.37 male/female. Flies were trapped throughout the year with peak population in summer. Peak activity was recorded in the afternoon (12.00–18.00h). In summary, the modified semi-automatic funnel fly trap can be used for field collection of the adult fly. By setting the timer, population dynamics, diversity, and periodic activity of adult flies were determined.

Senecio brasiliensis impairs eclosion rate and induces apoptotic cell death in larvae of Drosophila melanogaster

Senecio brasilienis (Spreng) Less., is a species native from Brazil, popularly known as “Maria mole”, and known to induce hepatotoxicity due to its high content of Pyrrolizidine alkaloids. Despite its toxicity, this plant is widely used in Brazilian folk medicine. Considering the antagonizing effects described for S. brasiliensis, we describe here molecular markers involved in the toxicity of hydroalcoholic extract from leaves of S. brasiliensis (HESB) in Drosophila melanogaster. Phytochemical analysis of HESB revealed the presence of phenolic acids and flavonoids. A significant antioxidant potential against ABTS+ and DPPH radical was found in parallel. Ingestion of extract did not alter the survival and locomotor activity of adult flies. However when ingested along the larval developmental phase, the eclosion rate of flies was interrupted at higher concentration of extract. To comprehend this phenomenon several analysis were conducted in larvae. HESB stimulated activity of antioxidant enzymes SOD and GST, and increased GSH/GSSG ratio and ROS production. Additionally, HESB caused a significant decrease of cell viability. The mRNA expression of Nrf2, TrxR, CAT, Drice and Dilp6 were also significantly up-regulated. HESB caused significant decrease on the phosphorylation of MAPKs and AKT. In parallel, PARP cleavage and caspases 3/7 activity were stimulated. In addition, glucose, glycogen and triglycerides levels were decreased. Taken together our study depicts a disruption in the eclosion of D. melanogaster possibly attributed to the inhibition of kinases implied in developmental process, energetic demand and induction of apoptotic cell death process.

Development of the Oriental Latrine Fly, Chrysomya megacephala (Diptera: Calliphoridae), at Five Constant Temperatures.

Chrysomya megacephala (Fabricius) is a forensically important fly that is found throughout the tropics and subtropics. We calculated the accumulated development time and transition points for each life stage from eclosion to adult emergence at five constant temperatures: 15, 20, 25, 30, and 35 °C. For each transition, the 10th, 50th, and 90th percentiles were calculated with a logistic linear model. The mean transition times and % survivorship were determined directly from the raw laboratory data. Development times of C. megacephala were compared with that of two other closely related species, Chrysomya rufifacies (Macquart) and Phormia regina (Meigen). Ambient and larval mass temperatures were collected from field studies conducted from 2001-2004. Field study data indicated that adult fly activity was reduced at lower ambient temperatures, but once a larval mass was established, heat generation occurred. These development times and durations can be used for estimation of a postmortem interval (PMI).

Modulatory Action by the Serotonergic System: Behavior and Neurophysiology in Drosophila melanogaster.

Serotonin modulates various physiological processes and behaviors. This study investigates the role of 5-HT in locomotion and feeding behaviors as well as in modulation of sensory-motor circuits. The 5-HT biosynthesis was dysregulated by feeding Drosophila larvae 5-HT, a 5-HT precursor, or an inhibitor of tryptophan hydroxylase during early stages of development. The effects of feeding fluoxetine, a selective serotonin reuptake inhibitor, during early second instars were also examined. 5-HT receptor subtypes were manipulated using RNA interference mediated knockdown and 5-HT receptor insertional mutations. Moreover, synaptic transmission at 5-HT neurons was blocked or enhanced in both larvae and adult flies. The results demonstrate that disruption of components within the 5-HT system significantly impairs locomotion and feeding behaviors in larvae. Acute activation of 5-HT neurons disrupts normal locomotion activity in adult flies. To determine which 5-HT receptor subtype modulates the evoked sensory-motor activity, pharmacological agents were used. In addition, the activity of 5-HT neurons was enhanced by expressing and activating TrpA1 channels or channelrhodopsin-2 while recording the evoked excitatory postsynaptic potentials (EPSPs) in muscle fibers. 5-HT2 receptor activation mediates a modulatory role in a sensory-motor circuit, and the activation of 5-HT neurons can suppress the neural circuit activity, while fluoxetine can significantly decrease the sensory-motor activity.

Development and validation of a new technique for estimating a minimum postmortem interval using adult blow fly (Diptera: Calliphoridae) carcass attendance.

Understanding the onset and duration of adult blow fly activity is critical to accurately estimating the period of insect activity or minimum postmortem interval (minPMI). Few, if any, reliable techniques have been developed and consequently validated for using adult fly activity to determine a minPMI. In this study, adult blow flies (Diptera: Calliphoridae) of Cochliomyia macellaria and Chrysomya rufifacies were collected from swine carcasses in rural central Texas, USA, during summer 2008 and Phormia regina and Calliphora vicina in the winter during 2009 and 2010. Carcass attendance patterns of blow flies were related to species, sex, and oocyte development. Summer-active flies were found to arrive 4-12 h after initial carcass exposure, with both C. macellaria and C. rufifacies arriving within 2 h of one another. Winter-active flies arrived within 48 h of one another. There was significant difference in degree of oocyte development on each of the first 3 days postmortem. These frequency differences allowed a minPMI to be calculated using a binomial analysis. When validated with seven tests using domestic and feral swine and human remains, the technique correctly estimated time of placement in six trials.

Extremely low frequency magnetic field (50 Hz, 0.5 mT) modifies fitness components and locomotor activity of Drosophila subobscura

Purpose: Extremely low frequency (ELF) magnetic fields are essential ecological factors which may induce changes in many organisms. The aim of this study was to examine the effects in Drosophila subobscura exposed for 48 h to ELF magnetic field (50 Hz, 0.5 mT) at different developmental stages.Materials and methods: Egg-first instar larvae developmental stage of D. subobscura isofemale lines was exposed to ELF magnetic field, and fitness components (developmental time, developmental dynamics, viability and sex ratio) and locomotor activity of three-day-old males and females were monitored. Also, just eclosed D. subobscura isofemale adults were exposed to ELF magnetic field and their locomotor activity was monitored just after.Results: ELF magnetic field shortens developmental time, increases viability and does not affect sex ratio of D. subobscura. No matter which developmental stage is exposed, ELF magnetic field significantly decreases locomotor activity of adult flies, but after exposure of just eclosed adults observed change lasts longer.Conclusions: Applied ELF magnetic field modifies fitness components and locomotor activity of D. subobscura. Observed effects can be attributed to the influence of magnetic field on different stages of development where the hormonal and nervous systems play important role in the control of examined parameters.

Clock accuracy and precision evolve as a consequence of selection for adult emergence in a narrow window of time in fruit flies Drosophila melanogaster

Although circadian clocks are believed to have evolved under the action of periodic selection pressures (selection on phasing) present in the geophysical environment, there is very little rigorous and systematic empirical evidence to support this. In the present study, we examined the effect of selection for adult emergence in a narrow window of time on the circadian rhythms of fruit flies Drosophila melanogaster. Selection was imposed in every generation by choosing flies that emerged during a 1 h window of time close to the emergence peak of baseline/control flies under 12 h:12 h light:dark cycles. To study the effect of selection on circadian clocks we estimated several quantifiable features that reflect inter- and intra-individual variance in adult emergence and locomotor activity rhythms. The results showed that with increasing generations, incidence of adult emergence and activity of adult flies during the 1 h selection window increased gradually in the selected populations. Flies from the selected populations were more homogenous in their clock period, were more coherent in their phase of entrainment, and displayed enhanced accuracy and precision in their emergence and activity rhythms compared with controls. These results thus suggest that circadian clocks in D. melanogaster evolve enhanced accuracy and precision when subjected to selection for emergence in a narrow window of time.

The Insulin-Regulated CREB Coactivator TORC Promotes Stress Resistance in Drosophila

In fasted mammals, glucose homeostasis is maintained through induction of the cAMP response element-binding protein (CREB) coactivator transducer of regulated CREB activity 2 (TORC2), which stimulates the gluconeogenic program in concert with the forkhead factor FOXO1. Here we show that starvation also triggers TORC activation in Drosophila, where it maintains energy balance through induction of CREB target genes in the brain. TORC mutant flies have reduced glycogen and lipid stores and are sensitive to starvation and oxidative stress. Neuronal TORC expression rescued stress sensitivity as well as CREB target gene expression in TORC mutants. During refeeding, increases in insulin signaling inhibited TORC activity through the salt-inducible kinase 2 (SIK2)-mediated phosphorylation and subsequent degradation of TORC. Depletion of neuronal SIK2 increased TORC activity and enhanced stress resistance. As disruption of insulin signaling also augmented TORC activity in adult flies, our results illustrate the importance of an insulin-regulated pathway that functions in the brain to maintain energy balance.

Opportunistic exploitation of turtle eggs by Tripanurga importuna (Walker) (Diptera: Sarcophagidae)

Tripanurga importuna (Walker, 1849) is a sarcophagid fly whose larvae often occur in nests of freshwater turtles. We investigated this sarcophagid fly to determine whether it is an opportunistic scavenger or a potential predator of eggs, embryos, and hatchlings of the spiny softshell turtle ( Apalone spinifera (Lesueur, 1827)). Fly infestation of spiny softshell turtle nests can occur at any time between oviposition and hatching, but estimates based on larval size and development time, along with observations of adult fly activity, suggest that female sarcophagids deposit larvae over the nest primarily during hatching. Observed temperature variance within the turtle clutch mass and physiological and developmental differences among eggs may result in asynchronous hatching, and chemical cues associated with early hatching may attract adult flies. Egg position within the nest affects embryo hatching success independently of fly infestation while also affecting fly infestation. Tripanurga importuna is a habitat specialist able to find and develop in carrion buried in sand, but it is a food opportunist able to develop on other buried carrion as well as turtle eggs. Tripanurga importuna maggots in turtle nests preferentially scavenge necrotic tissue, including damaged turtle eggs, but will opportunistically prey upon live embryos and hatchlings under some circumstances.

Tuning of RNA editing by ADAR is required in Drosophila

RNA editing increases during development in more than 20 transcripts encoding proteins involved in rapid synaptic neurotransmission in Drosophila central nervous system and muscle. Adar (adenosine deaminase acting on RNA) mutant flies expressing only genome-encoded, unedited isoforms of ion-channel subunits are viable but show severe locomotion defects. The Adar transcript itself is edited in adult wild-type flies to generate an isoform with a serine to glycine substitution close to the ADAR active site. We show that editing restricts ADAR function since the edited isoform of ADAR is less active in vitro and in vivo than the genome-encoded, unedited isoform. Ubiquitous expression in embryos and larvae of an Adar transcript that is resistant to editing is lethal. Expression of this transcript in embryonic muscle is also lethal, with above-normal, adult-like levels of editing at sites in a transcript encoding a muscle voltage-gated calcium channel.