Short and long-lasting behavioral consequences of agonistic encounters between male Drosophila melanogaster

In many animal species, learning and memory have been found to play important roles in regulating intra- and interspecific behavioral interactions in varying environments. In such contexts, aggression is commonly used to obtain desired resources. Previous defeats or victories during aggressive interactions have been shown to influence the outcome of later contests, revealing loser and winner effects. In this study, we asked whether short- and/or long-term behavioral consequences accompany victories and defeats in dyadic pairings between male Drosophila melanogaster and how long those effects remain. The results demonstrated that single fights induced important behavioral changes in both combatants and resulted in the formation of short-term loser and winner effects. These decayed over several hours, with the duration depending on the level of familiarity of the opponents. Repeated defeats induced a long-lasting loser effect that was dependent on de novo protein synthesis, whereas repeated victories had no long-term behavioral consequences. This suggests that separate mechanisms govern the formation of loser and winner effects. These studies aim to lay a foundation for future investigations exploring the molecular mechanisms and circuitry underlying the nervous system changes induced by winning and losing bouts during agonistic encounters.Across the animal kingdom, aggression between conspecifics often accompanies the competition for food, mates, and territory. Although an innate behavior, aggression is a highly adaptive trait as well, with animals learning from previous experience and changing their behavior in response to new challenges. In competition for rank, for example, previous fighting experience influences the outcome of subsequent contests: prior defeat decreases whereas prior victory increases the probability of winning later contests. These have been called “loser” and “winner” effects (1). Such effects have been observed in many species, including fish (2), birds (3), and mammals (4). In general, loser effects persist longer than winner effects (5). The durational asymmetry observed between loser and winner effects has been hypothesized to participate in stabilizing social hierarchies among conspecifics (6).Fruit flies (Drosophila melanogaster) exhibit a variety of simple and complex social behaviors, including aggregation (7), courtship (8), and aggression (9) in which learning and memory have been demonstrated or postulated to serve important roles (10–12). Thus, characterizing the molecular basis of memory formation, retention, and retrieval is crucial to ultimately understanding the adaptability of these social behaviors. In Drosophila, a variety of operant and classical training paradigms have been used to subdivide memory into distinct categories. Short-term memory (STM) lasting minutes to hours is induced by a single training session, whereas long-term memory (LTM) lasting days usually requires repeated training sessions and involves de novo protein synthesis (13). A large number of studies have been carried out using olfactory, visual, social, and place memory paradigms. These have allowed the functional and molecular characterization of neuronal circuits and the identification of numerous genes underlying learning and memory (14–16). Included are mutations in rutabaga (rut, type 1 adenylyl cyclase) that interfere with learning and STM formation (17); amnesiac (amn, peptide regulator of adenylyl cyclase) that affect STM retention (18); and crammer (cer, inhibitor of a cathepsin subfamily) that prevent LTM formation (19). Whether rut, amn, and cer serve roles in the learning and memory that accompanies aggression remains unknown.Male–male aggression in fruit flies was first described almost 100 y ago (20). Since then, considerable progress has been made in understanding its expression and regulation (21–26). In competition for food resources and territory, male–male agonistic encounters, composed of stereotyped behavioral patterns, usually result in the formation of dominance relationships (9). During the progression of fights, both flies modify their fighting strategies: The ultimate winners chase and lunge at their opponents to gain sole access to the resources, whereas the losers retreat from the resources after receiving such attacks (9, 10).In second fights (30 min after first fights), losing flies display greater submissive behavior and never win against naïve or experienced opponents, revealing short-term loser effects (10). Evidence for winner effects, however, was not found (11). Recently, in olive fruit flies (Bactrocera olea) it was found that previous losing and winning experiences both increased the aggressiveness of the flies. This suggests that the consequences of losing or winning may vary across species (27).We previously suggested that fights between male flies function as operant learning situations in which males learn to use their most advantageous fighting strategy during fights and then continue to do so in subsequent contests (28). In an attempt to optimize the learning and memory associated with aggression, we designed new “handling-free” behavioral chambers (29). These proved to be more desirable for studying the formation of loser effects (12). By using these experimental arenas and pairing familiar opponents in second fights we previously showed that changes in fighting strategies could be developed by both winning and losing flies. This allowed us to suggest the existence of short-term winner effects along with the previously demonstrated loser effects (12). A more detailed examination of these short-term effects is presented here along with experiments attempting to measure the intrinsic changes in fighting abilities of losing and winning flies.In the present study, we ask (i) whether a single fight can lead to the formation of loser and winner effects and how long these effects persist, (ii) whether flies adopt different fighting strategies in second fights depending on their opponents, (iii) whether longer-lasting behavioral effects result from sequential repeated defeats or victories, (iv) whether protein synthesis is required for the short- or long-term effects observed, and (v) whether mutations in genes involved in learning and memory affect aggressive behavior.     

In between: Gypsy in Drosophila melanogaster Reveals New Insights into Endogenous Retrovirus Evolution

Retroviruses are RNA viruses that are able to synthesize a DNA copy of their genome and insert it into a chromosome of the host cell. Sequencing of different eukaryote genomes has revealed the presence of many such endogenous retroviral sequences. The mechanisms by which these retroviral sequences have colonized the genome are still unknown, and the endogenous retrovirus gypsy of Drosophila melanogaster is a powerful experimental model for deciphering this process in vivo. Gypsy is expressed in a layer of somatic cells, and then transferred into the oocyte by an unknown mechanism. This critical step is the start of the endogenization process. Moreover gypsy has been shown to have infectious properties, probably due to its envelope gene acquired from a baculovirus. Recently we have also shown that gypsy maternal transmission is reduced in the presence of the endosymbiotic bacterium Wolbachia. These studies demonstrate that gypsy is a unique and powerful model for understanding the endogenization of retroviruses.     

A search for principles of disability using experimental impairment of Drosophila melanogaster

The results of life table experiments to determine the effects of artificial impairment (leg amputation) in 7500 Drosophila melanogaster adults revealed that the extent to which life expectancy was reduced in impaired individuals was conditional on: (1) leg location and number amputated--front leg had greatest impact and the number of legs amputated directly correlated with mortality impact; (2) age of amputation--the greatest relative reduction in remaining life expectancy occurred when young flies were impaired; (3) vial orientation--mortality in impaired flies was the least when vials held upside-down (most friendly environment) and the greatest when they were right-side up (least friendly environment); and (4) sex--male mortality was increased more than female mortality in nearly all impairment treatments. These results were used to formulate a set of general principles of disability that would apply not only to humans but to all organisms.     

Decanalization of wing development accompanied the evolution of large wings in high-altitude Drosophila

In higher organisms, the phenotypic impacts of potentially harmful or beneficial mutations are often modulated by complex developmental networks. Stabilizing selection may favor the evolution of developmental canalization—that is, robustness despite perturbation—to insulate development against environmental and genetic variability. In contrast, directional selection acts to alter the developmental process, possibly undermining the molecular mechanisms that buffer a trait’s development, but this scenario has not been shown in nature. Here, we examined the developmental consequences of size increase in highland Ethiopian Drosophila melanogaster. Ethiopian inbred strains exhibited much higher frequencies of wing abnormalities than lowland populations, consistent with an elevated susceptibility to the genetic perturbation of inbreeding. We then used mutagenesis to test whether Ethiopian wing development is, indeed, decanalized. Ethiopian strains were far more susceptible to this genetic disruption of development, yielding 26 times more novel wing abnormalities than lowland strains in F2 males. Wing size and developmental perturbability cosegregated in the offspring of between-population crosses, suggesting that genes conferring size differences had undermined developmental buffering mechanisms. Our findings represent the first observation, to our knowledge, of morphological evolution associated with decanalization in the same tissue, underscoring the sensitivity of development to adaptive change.Canalization describes the property of some biological traits to remain constant in the face of environmental and genetic changes (1–6). This phenomenon has important implications for the relationship between genetic and phenotypic variation. By masking the phenotypic effects of genetic changes, canalization may inhibit phenotypic evolution while allowing hidden genetic variation to accumulate. If canalization is overcome by environmental and/or genetic changes, this reservoir of functional variation may then be exposed. For example, Waddington (7) selected Drosophila for a missing cross-vein trait that initially only appeared in a stressful high-temperature environment but after selection, manifested under normal conditions as well. A molecular case study of canalization was provided by Rutherford and Lindquist (8), who found that Drosophila with a disabled chaperone protein (the heat shock protein Hsp90) showed a suite of developmental abnormalities. These abnormalities varied based on genetic background and environment and could be selected for Hsp90 independence. Other studies have also found that selection in the laboratory can alter developmental stability (9–12), and Hayden et al. (13) found that in vitro directional selection on ribozyme activity led to reduced genetic and environmental robustness.Canalization is difficult to disentangle from selective constraint, which complicates its study in natural populations. A rare potential example comes from the blowfly Lucilia cuprina, in which the evolution of insecticide resistance was accompanied by prolonged development and bristle asymmetry (14). Those disadvantages were subsequently reversed by the evolution of an unlinked modifier locus (15). Here, the initial cost of adaptation may have been because of pleiotropic decanalizing effects of the insecticide resistance mutation itself. Or given the contrast between the adaptive and decanalized phenotypes, a linked deleterious variant might have been fixed along with the resistance allele and later compensated by the modifier gene.Canalization might evolve because of stabilizing selection favoring the same phenotypic optimum in the face of environmental and genetic variability [as shown in the case of environmental robustness (16)], or canalization might arise from inherent properties of the biological system (17). Particularly in the former scenario, it seems possible that directional selection might undermine canalization: if selection for a new phenotypic optimum alters the developmental process, then the molecular mechanisms that had previously buffered the ancestral phenotype might fail to buffer the novel phenotype. Hence, it seems possible that recently evolved traits may show reduced canalization (until new or modified buffering mechanisms can evolve), but no such example has been reported from nature.Here, we describe a natural instance of decanalization associated with a recently evolved morphological structure, focusing on wing size and developmental stability in a high-altitude (>3,000 m) Ethiopian population of Drosophila melanogaster. Although globally distributed today, this human-commensal species probably originated in the lowlands of southern central Africa (18). The species’ arrival in Ethiopia may have roughly coincided with its crossing of the Sahara [∼10,000 y ago (19, 20)]. Highland Ethiopian flies are morphologically divergent from other D. melanogaster populations, featuring striking melanism (21), larger body size, and larger wings (Fig. 1) with distinct shape (22).Open in a separate windowFig. 1.Morphological comparisons of D. melanogaster from the Ethiopian highlands and an ancestral range Zambia population. Ethiopian strains have (A vs. B) larger body size and (C vs. D) wing size. (E) The distribution of wing widths among outbred individuals shows almost no overlap between populations. Detailed size data are given in Dataset S1.Past studies have shown that the Drosophila wing provides a convenient visible readout of development, allowing, for example, the study of variation unmasked by specific mutations (23, 24). This study repurposes mutagenesis as a generalized genetic perturbation to assess whether wing size evolution in Ethiopian D. melanogaster has undermined the stability of wing development. Initially, our observation of frequent wing abnormalities in Ethiopian inbred strains motivated the hypothesis of decanalized wing development. Mutagenesis experiments confirmed that de novo mutations were far more likely to produce wing defects in the Ethiopian strains than in the smaller-winged Zambia population (whereas a control trait showed no such difference), implying less buffered development of Ethiopian wings. A final mutagenesis experiment confirmed that wing size and decanalization were inherited together in the advanced generation offspring of an Ethiopia–Zambia cross, implying that alleles conferring larger Ethiopian wings contributed to destabilized development.     

Tissue nonautonomous effects of fat body methionine metabolism on imaginal disc repair in Drosophila

Regulatory mechanisms for tissue repair and regeneration within damaged tissue have been extensively studied. However, the systemic regulation of tissue repair remains poorly understood. To elucidate tissue nonautonomous control of repair process, it is essential to induce local damage, independent of genetic manipulations in uninjured parts of the body. Herein, we develop a system in Drosophila for spatiotemporal tissue injury using a temperature-sensitive form of diphtheria toxin A domain driven by the Q system to study factors contributing to imaginal disc repair. Using this technique, we demonstrate that methionine metabolism in the fat body, a counterpart of mammalian liver and adipose tissue, supports the repair processes of wing discs. Local injury to wing discs decreases methionine and S-adenosylmethionine, whereas it increases S-adenosylhomocysteine in the fat body. Fat body-specific genetic manipulation of methionine metabolism results in defective disc repair but does not affect normal wing development. Our data indicate the contribution of tissue interactions to tissue repair in Drosophila, as local damage to wing discs influences fat body metabolism, and proper control of methionine metabolism in the fat body, in turn, affects wing regeneration.Tissue repair is the necessary ability of an organism to maintain tissue homeostasis. The self-repair mechanisms of injured tissues have been extensively studied (1, 2). The homeostatic mechanisms of tissue repair, on the other hand, are not confined to the damaged tissues, but rather involve organismal regulation with contributions from multiple systems, including the circulatory, nervous, and endocrine systems. The importance of interactions between damaged tissue and other tissues has been suggested. For example, nerve-derived factor is essential for limb regeneration in the newt and axolotl (3, 4). Studies of parabiosis, the fusion of two organisms that consequently share a vascular system, revealed that regenerative ability was affected by internal environmental conditions based on the circulation of body fluids (5, 6). However, the molecular mechanisms of tissue nonautonomous regulation of the repair process are only beginning to be understood.Drosophila is a useful model animal for studying tissue interactions because of the availability of tissue-specific gene manipulation systems, such as the Gal4/Upstream Activation Sequence (UAS) system (7) or the Q system (8, 9). In Drosophila larvae, the epithelial sheets of imaginal discs are known to have a remarkable ability to repair massive tissue damage. A classic example is their regenerative capacity following surgical ablation (10–12). To overcome the technical difficulties of surgical ablation/regeneration experiments, a system was established for studying imaginal discs repair following genetic ablation, using a Gal4/UAS/Gal80^ts system (13). Despite the great advantages provided by this system, the use of Gal80^ts limits the manipulation of genes with UAS constructs to only the ablating cells at the time of ablation. One strategy for overcoming this problem is to conduct tissue ablation independent of the Gal4/UAS/Gal80^ts system, which allows for utilization of the Gal4/UAS system to manipulate gene expression in uninjured parts of the body. In the present study, we established a cell ablation system using a temperature-sensitive form of the diphtheria toxin A domain (DtA^ts) (14). It has been demonstrated that DtA^ts is active at low temperatures (18 °C) and induces cell death through nuclease activity, but is inactivated at high temperatures (29 °C) (14, 15). Inducing DtA^ts expression by using the Q system enabled us to manipulate Gal4/UAS-mediated gene expression in other organs, independent of the temporal tissue damage in wing discs.Several studies using Drosophila have demonstrated that damaged tissues send signals to surrounding tissues. Cytokine signaling from UV-damaged epidermal cells mediates nociceptive sensitization in larvae (16). Disc injury caused by aseptic wounds leads to production of Unpaired 3 (Upd3), an IL-6 like cytokine in the hemocytes and fat body, which triggers hemocyte proliferation, probably to accelerate clearance of injured cells (17). Hemocyte ablation, however, had no effect on wound closure in the injured larval epidermis (18), raising the question in terms of the functional contribution of hemocytes to tissue repair. In addition, damaged imaginal discs secrete Drosophila insulin-like peptide 8 (Dilp8), which inhibits ecdysone biosynthesis in the ring gland and arrests developmental processes, probably to gain time for repair (19, 20).We previously revealed that local necrosis in adult wings resulted in reduced levels of systemic S-adenosylmethionine (SAM) through the up-regulation of glycine N-methyltransferase (gnmt) in the fat body (21). This report is an example of systemic regulation of SAM metabolism in the fat body on epithelial damage. SAM is a metabolite present in all living cells; it plays various roles, including that of a precursor for transmethylation, transsulfuration, and polyamine biosynthesis (22). It is produced by sole SAM synthase (Sams) from the essential amino acid methionine (Met) and ATP, and SAM levels are strongly regulated by Gnmt in Drosophila fat body (21, 22). Gnmt consumes excess SAM and produces S-adenosylhomocysteine (SAH), which is further metabolized to homocysteine (Hcy). Changes in SAM levels in response to necrotic tissue led us to hypothesize that tissue damage in larval discs also affects methionine metabolism in the fat body, which turned out to be the case in the present study using DtA^ts-ablation system. We further tested whether the modulation of methionine metabolism in the fat body could affect imaginal disc repair in a tissue nonautonomous manner. Our study indicated that proper control of methionine metabolism in the fat body is crucial for repair of wing discs, highlighting the significance of systemic regulation for epithelial tissue repair.     

Dissecting neural pathways for forgetting in Drosophila olfactory aversive memory

Recent studies have identified molecular pathways driving forgetting and supported the notion that forgetting is a biologically active process. The circuit mechanisms of forgetting, however, remain largely unknown. Here we report two sets of Drosophila neurons that account for the rapid forgetting of early olfactory aversive memory. We show that inactivating these neurons inhibits memory decay without altering learning, whereas activating them promotes forgetting. These neurons, including a cluster of dopaminergic neurons (PAM-β′1) and a pair of glutamatergic neurons (MBON-γ4>γ1γ2), terminate in distinct subdomains in the mushroom body and represent parallel neural pathways for regulating forgetting. Interestingly, although activity of these neurons is required for memory decay over time, they are not required for acute forgetting during reversal learning. Our results thus not only establish the presence of multiple neural pathways for forgetting in Drosophila but also suggest the existence of diverse circuit mechanisms of forgetting in different contexts.Although forgetting commonly has a negative connotation, it is a functional process that shapes memory and cognition (1–4). Recent studies, including work in relatively simple invertebrate models, have started to reveal basic biological mechanisms underlying forgetting (5–15). In Drosophila, single-session Pavlovian conditioning by pairing an odor (conditioned stimulus, CS) with electric shock (unconditioned stimulus, US) induces aversive memories that are short-lasting (16). The memory performance of fruit flies is observed to drop to a negligible level within 24 h, decaying rapidly early after training and slowing down thereafter (17). Memory decay or forgetting requires the activation of the small G protein Rac, a signaling protein involved in actin remodeling, in the mushroom body (MB) intrinsic neurons (6). These so-called Kenyon cells (KCs) are the neurons that integrate CS–US information (18, 19) and support aversive memory formation and retrieval (20–22). In addition to Rac, forgetting also requires the DAMB dopamine receptor (7), which has highly enriched expression in the MB (23). Evidence suggests that the dopamine-mediated forgetting signal is conveyed to the MB by dopamine neurons (DANs) in the protocerebral posterior lateral 1 (PPL1) cluster (7, 24). Therefore, forgetting of olfactory aversive memory in Drosophila depends on a particular set of intracellular molecular pathways within KCs, involving Rac, DAMB, and possibly others (25), and also receives modulation from extrinsic neurons. Although important cellular evidence supporting the hypothesis that memory traces are erased under these circumstances is still lacking, these findings lend support to the notion that forgetting is an active, biologically regulated process (17, 26).Although existing studies point to the MB circuit as essential for forgetting, several questions remain to be answered. First, whereas the molecular pathways for learning and forgetting of olfactory aversive memory are distinct and separable (6, 7), the neural circuits seem to overlap. Rac-mediated forgetting has been localized to a large population of KCs (6), including the γ-subset, which is also critical for initial memory formation (21, 27). The site of action of DAMB for forgetting has yet to be established; however, the subgroups of PPL1-DANs implicated in forgetting are the same as those that signal aversive reinforcement and are required for learning (28–30). It leaves open the question of whether the brain circuitry underlying forgetting and learning is dissociable, or whether forgetting and learning share the same circuit but are driven by distinct activity patterns and molecular machinery (26). Second, shock reinforcement elicits multiple memory traces through at least three dopamine pathways to different subdomains in the MB lobes (28, 29). Functional imaging studies have also revealed Ca²⁺-based memory traces in different KC populations (31). It is poorly understood how forgetting of these memory traces differs, and it remains unknown whether there are multiple regulatory neural pathways. Notably, when PPL1-DANs are inactivated, forgetting still occurs, albeit at a lower rate (7). This incomplete block suggests the existence of an additional pathway(s) that conveys forgetting signals to the MB. Third, other than memory decay over time, forgetting is also observed through interference (32, 33), when new learning or reversal learning is introduced after training (6, 34, 35). Time-based and interference-based forgetting shares a similar dependence on Rac and DAMB (6, 7). However, it is not known whether distinct circuits underlie forgetting in these different contexts.In the current study, we focus on the diverse set of MB extrinsic neurons (MBENs) that interconnect the MB lobes with other brain regions, which include 34 MB output neurons (MBONs) of 21 types and ∼130 dopaminergic neurons of 20 types in the PPL1 and protocerebral anterior medial (PAM) clusters (36, 37). These neurons have been intensively studied in olfactory memory formation, consolidation, and retrieval in recent years (e.g., 24, 28–30, 38–48); however, their roles in forgetting have not been characterized except for the aforementioned PPL1-DANs. In a functional screen, we unexpectedly found that several Gal4 driver lines of MBENs showed significantly better 3-h memory retention when the Gal4-expressing cells were inactivated. The screen has thus led us to identify two types of MBENs that are not involved in initial learning but play important and additive roles in mediating memory decay. Furthermore, neither of these MBEN types is required for reversal learning, supporting the notion that there is a diversity of neural circuits that drive different forms of forgetting.     

Effect of X-irradiation at larval stage on adult lifespan in Drosophila melanogaster

The effects of X-ray irradiation at larval stage with doses of 1.2, 2.1, 4.2, 7.5 and 17.1 Gy on adult longevity and fecundity in Drosophila melanogaster fruit flies were studied. A significant negative trend with increasing dose of irradiation was detected for the median lifespan in both sexes. In all experimental groups, both male and female mortality rates in irradiated flies were above control levels approximately for one month after emergence, and below control levels at older ages. The irradiation with 1.2 and 2.1 Gy resulted in 11.5% and 12.7% increase of male maximum lifespan, respectively. Irradiated females had in most cases a lower fecundity than control females. In all studied age groups, the decrease of fecundity was dose-related, and the negative effect of irradiation on fecundity was no longer observed in flies older than two weeks of age. Mean fecundity for the 4-25-day period of the irradiated females was shortened and dose-related [one-way ANOVA: F(5,414) = 10.56, P < 0.001], but significant differences from control were observed only for flies irradiated with doses of 4.2, 7.5 and 17.1 Gy. Mean fecundity for females irradiated with doses of 1.2 and 2.1 Gy did not differ from that of control females.     

Profiling early infection responses: Pseudomonas aeruginosa eludes host defenses by suppressing antimicrobial peptide gene expression

Insights into the host factors and mechanisms mediating the primary host responses after pathogen presentation remain limited, due in part to the complexity and genetic intractability of host systems. Here, we employ the model Drosophila melanogaster to dissect and identify early host responses that function in the initiation and progression of Pseudomonas aeruginosa pathogenesis. First, we use immune potentiation and genetic studies to demonstrate that flies mount a heightened defense against the highly virulent P. aeruginosa strain PA14 when first inoculated with strain CF5, which is avirulent in flies; this effect is mediated via the Imd and Toll signaling pathways. Second, we use whole-genome expression profiling to assess and compare the Drosophila early defense responses triggered by the PA14 vs. CF5 strains to identify genes whose expression patterns are different in susceptible vs. resistant host-pathogen interactions, respectively. Our results identify pathogenesis- and defense-specific genes and uncover a previously undescribed mechanism used by P. aeruginosa in the initial stages of its host interaction: suppression of Drosophila defense responses by limiting antimicrobial peptide gene expression. These results provide insights into the genetic factors that mediate or restrict pathogenesis during the early stages of the bacterial-host interaction to advance our understanding of P. aeruginosa-human infections.     

Mechanical feedback as a possible regulator of tissue growth

Regulation of cell growth and proliferation has a fundamental role in animal and plant development and in the progression of cancer. In the context of development, it is important to understand the mechanisms that coordinate growth and patterning of tissues. Imaginal discs, which are larval precursors of fly limbs and organs, have provided much of what we currently know about these processes. Here, we consider the mechanism that is responsible for the observed uniformity of growth in wing imaginal discs, which persists in the presence of gradients in growth inducing morphogens in spite of the stochastic nature of cell division. The phenomenon of “cell competition,” which manifests in apoptosis of slower-growing cells in the vicinity of faster growing tissue, suggests that uniform growth is not a default state but a result of active regulation. How can a patch of tissue compare its growth rate with that of its surroundings? A possible way is furnished by mechanical interactions. To demonstrate this mechanism, we formulate a mathematical model of nonuniform growth in a layer of tissue and examine its mechanical implications. We show that a clone growing faster or slower than the surrounding tissue is subject to mechanical stress, and we propose that dependence of the rate of cell division on local stress could provide an “integral-feedback” mechanism stabilizing uniform growth. The proposed mechanism of growth control is not specific to imaginal disc growth and could be of general relevance. Several experimental tests of the proposed mechanism are suggested.     

Redundant mechanisms mediate bristle patterning on the Drosophila thorax

The thoracic bristle pattern of Drosophila results from the spatially restricted expression of the achaete-scute (ac-sc) genes in clusters of cells, mediated by the activity of many discrete cis-regulatory sequences. However, ubiquitous expression of sc or asense (ase) achieved with a heterologous promoter, in the absence of endogenous ac-sc expression, and the activity of the cis-regulatory elements, allows the development of bristles positioned at wild-type locations. We demonstrate that the products of the genes stripe, hairy, and extramacrochaetae contribute to rescue by antagonizing the activity of Sc and Ase. The three genes are expressed in specific but overlapping spatial domains of expression that form a prepattern that allows precise positioning of bristles. The redundant mechanisms might contribute to the robustness of the pattern. We discuss the possibility that patterning in trans by antagonism is ancestral and that the positional cis-regulatory sequences might be of recent origin.     

Commensal bacteria play a role in mating preference of Drosophila melanogaster

Development of mating preference is considered to be an early event in speciation. In this study, mating preference was achieved by dividing a population of Drosophila melanogaster and rearing one part on a molasses medium and the other on a starch medium. When the isolated populations were mixed, "molasses flies" preferred to mate with other molasses flies and "starch flies" preferred to mate with other starch flies. The mating preference appeared after only one generation and was maintained for at least 37 generations. Antibiotic treatment abolished mating preference, suggesting that the fly microbiota was responsible for the phenomenon. This was confirmed by infection experiments with microbiota obtained from the fly media (before antibiotic treatment) as well as with a mixed culture of Lactobacillus species and a pure culture of Lactobacillus plantarum isolated from starch flies. Analytical data suggest that symbiotic bacteria can influence mating preference by changing the levels of cuticular hydrocarbon sex pheromones. The results are discussed within the framework of the hologenome theory of evolution.     

Bombykol receptors in the silkworm moth and the fruit fly

Male moths are endowed with odorant receptors (ORs) to detect species-specific sex pheromones with remarkable sensitivity and selectivity. We serendipitously discovered that an endogenous OR in the fruit fly, Drosophila melanogaster, is highly sensitive to the sex pheromone of the silkworm moth, bombykol. Intriguingly, the fruit fly detectors are more sensitive than the receptors of the silkworm moth, although its ecological significance is unknown. By expression in the “empty neuron” system, we identified the fruit fly bombykol-sensitive OR as DmelOR7a (= DmOR7a). The profiles of this receptor in response to bombykol in the native sensilla (ab4) or expressed in the empty neuron system (ab3 sensilla) are indistinguishable. Both WT and transgenic flies responded with high sensitivity, in a dose-dependent manner, and with rapid signal termination. In contrast, the same empty neuron expressing the moth bombykol receptor, BmorOR1, demonstrated low sensitivity and slow signal inactivation. When expressed in the trichoid sensilla T1 of the fruit fly, the neuron housing BmorOR1 responded with sensitivity comparable to that of the native trichoid sensilla in the silkworm moth. By challenging the native bombykol receptor in the fruit fly with high doses of another odorant to which the receptor responds with the highest sensitivity, we demonstrate that slow signal termination is induced by overdose of a stimulus. As opposed to the empty neuron system in the basiconic sensilla, the structural, biochemical, and/or biophysical features of the sensilla make the T1 trichoid system of the fly a better surrogate for the moth receptor.     

Cu, Zn superoxide dismutase deficiencyaccelerates the time course of an age-related markerin Drosophila melanogaster

In the oxidative stress hypothesis of aging therandom accumulation of oxidative damage over time ispostulated to cause aging. The pace at whichoxidative damage accrues determines the rate of aging,but it is less clear how the accumulation of randomdamage could cause the stereotypic pattern of aging. It has been proposed that oxidative damage induceschanges in gene expression, translating a random inputof damage into a patterned output. In support of thiswe show that in adult Drosophila melanogaster,with a deficiency in the anti-oxidant enzyme Cu, Znsuperoxide dismutase (Sod), an increase in oxidativestress, and a shortened life span, there isacceleration in the normal age-related temporalpattern of wingless gene expression. Theacceleration in the temporal pattern of winglessgene expression is proportional to the shortened lifespan suggesting that the shortened life span of Soddeficient animals is due, not to an abnormalpathological process, but to an increase in the rateof aging.     

Drosophila melanogaster as a model host to dissect the immunopathogenesis of zygomycosis

Zygomycosis is an emerging frequently fatal opportunistic mycosis whose immunopathogenesis is poorly understood. We developed a zygomycosis model by injecting Drosophila melanogaster flies with a standardized amount of fungal spores from clinical Zygomycetes isolates to study virulence and host defense mechanisms. We found that, as opposed to most other fungi, which are nonpathogenic in D. melanogaster (e.g., Aspergillus fumigatus), Zygomycetes rapidly infect and kill wild-type flies. Toll-deficient flies exhibited increased susceptibility to Zygomycetes, whereas constitutive overexpression of the antifungal peptide Drosomycin in transgenic flies partially restored resistance to zygomycosis. D. melanogaster Schneider 2 phagocytic cells displayed decreased phagocytosis and caused less hyphal damage to Zygomycetes compared with that to A. fumigatus. Furthermore, phagocytosis-defective eater mutant flies displayed increased susceptibility to Zygomycetes infection. Classic enhancers of Zygomycetes virulence in humans, such as corticosteroids, increased iron supply, and iron availability through treatment with deferoxamine dramatically increased Zygomycetes pathogenicity in our model. In contrast, iron starvation induced by treatment with the iron chelator deferasirox significantly protected flies infected with Zygomycetes. Whole-genome expression profiling in wild-type flies after infection with Zygomycetes vs. A. fumigatus identified genes selectively down-regulated by Zygomycetes, which act in pathogen recognition, immune defense, stress response, detoxification, steroid metabolism, or tissue repair or have unknown functions. Our results provide insights into the factors that mediate host-pathogen interactions in zygomycosis and establish D. melanogaster as a promising model to study this important mycosis.