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Commensal microbes influence various aspects of vertebrate and invertebrate brain function. We previously reported that Lactiplantibacillus plantarum SBT2227 promotes sleep in the fruit fly, Drosophila melanogaster. However, how widely the sleep-promoting effects are conserved in gut bacterial species remains unknown. In this study, we orally administered human intestinal and food-associated bacterial species (39 in total) to flies and investigated their effects on sleep. Six species of bacteria were found to have significant sleep-promoting effects. Of these, we further investigated Bifidobacterium adolescentis, which had the greatest sleep-promoting effect, and found that the strength of the sleep effect varied among strains of the same bacterial species. The B. adolescentis strains BA2786 and BA003 showed strong and weak effects on sleep, respectively. Transcriptome characteristics compared between the heads of flies treated with BA2786 or BA003 revealed that the gene expression of the insulin-like receptor (InR) was increased in BA2786-fed flies. Furthermore, a heterozygous mutation in InR suppressed the sleep-promoting effect of BA2786. These results suggest that orally administered sleep-promoting bacteria (at least BA2786), may act on insulin signaling to modulate brain function for sleep.
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Drosophila melanogaster , Sono , Animais , Humanos , Drosophila melanogaster/genética , Sono/genética , Bactérias , InsulinaRESUMO
During courtship, multiple information sources are integrated in the brain to reach a final decision, i.e., whether or not to mate. The brain functions for this complex behavior can be investigated by genetically manipulating genes and neurons, and performing anatomical, physiological, and behavioral analyses. Drosophila is a powerful model experimental system for such studies, which need to be integrated from molecular and cellular levels to the behavioral level, and has enabled pioneering research to be conducted. In male flies, which exhibit a variety of characteristic sexual behaviors, we have accumulated knowledge of many genes and neural circuits that control sexual behaviors. On the other hand, despite the importance of the mechanisms of mating decision-making in females from an evolutionary perspective (such as sexual selection), research on the mechanisms that control sexual behavior in females has progressed somewhat slower. In this review, we focus on the pre-mating behavior of female Drosophila melanogaster, and introduce previous key findings on the neuronal and molecular mechanisms that integrate sensory information and selective expression of behaviors toward the courting male.
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Drosophila melanogaster/fisiologia , Motivação/fisiologia , Neurônios/fisiologia , Comportamento Sexual Animal/fisiologia , Animais , Feminino , HumanosRESUMO
Many animals use acoustic signals to attract a potential mating partner. In fruit flies (Drosophila melanogaster), the courtship pulse song has a species-specific interpulse interval (IPI) that activates mating. Although a series of auditory neurons in the fly brain exhibit different tuning patterns to IPIs, it is unclear how the response of each neuron is tuned. Here, we studied the neural circuitry regulating the activity of antennal mechanosensory and motor center (AMMC)-B1 neurons, key secondary auditory neurons in the excitatory neural pathway that relay song information. By performing Ca2+ imaging in female flies, we found that the IPI selectivity observed in AMMC-B1 neurons differs from that of upstream auditory sensory neurons [Johnston's organ (JO)-B]. Selective knock-down of a GABAA receptor subunit in AMMC-B1 neurons increased their response to short IPIs, suggesting that GABA suppresses AMMC-B1 activity at these IPIs. Connection mapping identified two GABAergic local interneurons that synapse with AMMC-B1 and JO-B. Ca2+ imaging combined with neuronal silencing revealed that these local interneurons, AMMC-LN and AMMC-B2, shape the response pattern of AMMC-B1 neurons at a 15 ms IPI. Neuronal silencing studies further suggested that both GABAergic local interneurons suppress the behavioral response to artificial pulse songs in flies, particularly those with a 15 ms IPI. Altogether, we identified a circuit containing two GABAergic local interneurons that affects the temporal tuning of AMMC-B1 neurons in the song relay pathway and the behavioral response to the courtship song. Our findings suggest that feedforward inhibitory pathways adjust the behavioral response to courtship pulse songs in female flies.SIGNIFICANCE STATEMENT To understand how the brain detects time intervals between sound elements, we studied the neural pathway that relays species-specific courtship song information in female Drosophila melanogaster We demonstrate that the signal transmission from auditory sensory neurons to key secondary auditory neurons antennal mechanosensory and motor center (AMMC)-B1 is the first-step to generate time interval selectivity of neurons in the song relay pathway. Two GABAergic local interneurons are suggested to shape the interval selectivity of AMMC-B1 neurons by receiving auditory inputs and in turn providing feedforward inhibition onto AMMC-B1 neurons. Furthermore, these GABAergic local interneurons suppress the song response behavior in an interval-dependent manner. Our results provide new insights into the neural circuit basis to adjust neuronal and behavioral responses to a species-specific communication sound.
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Drosophila melanogaster/fisiologia , Interneurônios/fisiologia , Comportamento Sexual Animal/fisiologia , Vocalização Animal/fisiologia , Ácido gama-Aminobutírico/fisiologia , Animais , Antenas de Artrópodes/fisiologia , Sinalização do Cálcio , Copulação , Feminino , Mecanorreceptores/fisiologia , Vias Neurais/fisiologia , Neurônios/fisiologia , Receptores de GABA-A/fisiologiaRESUMO
Many animals utilize auditory signals to communicate with conspecific individuals. During courtship, males of the fruit fly Drosophila melanogaster and related species produce a courtship song comprised of sine and pulse songs by vibrating their wings. The pulse song increases female receptivity and male courtship activity, indicating that it functions as a sexual signal. One song parameter, interpulse interval (IPI), varies among closely related species. In D. melanogaster, a song with a conspecific IPI induces a stronger behavioral response than heterospecific songs, indicating the ability of the flies to discriminate conspecific IPI. Traditionally, the fly's response to the song is measured under grouped conditions, in which the effect of sensory modalities other than audition cannot be excluded. Here, to quantify the individual ability to discriminate a conspecific song, we systematically analyzed the auditory response of single male flies to sound with various parameters. Moreover, we applied this method, termed SMART (Single Male Auditory Response Test), to two sister species for potential application in a comparative approach. By quantifying the locomotor activity of single D. melanogaster males during sound exposure, we detected increased locomotor activity in response to pulse songs, but not to white noise or pure tone. The conspecific song evoked stronger response than the heterospecific songs, and ablation of their antennal receivers severely suppressed the locomotor increase. A pulse song with a small IPI variation evoked a continuous response, while the response to songs with highly variable IPIs tends to be rapidly decayed. This provides the first evidence that fruit flies discriminate IPI variations, which possibly inform the age and social contexts of the singer. Sister species, D. sechellia, exhibited a locomotor response to pulse song, while D. simulans exhibited no behavioral response. This suggests that auditory and other stimuli that elicit this behavioral response are diversified among Drosophila species.
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Comunicação Animal , Percepção Auditiva/fisiologia , Comportamento Animal/fisiologia , Drosophila melanogaster/fisiologia , Estimulação Acústica , Animais , Corte , Locomoção/fisiologia , Masculino , Comportamento Sexual Animal/fisiologiaRESUMO
Mechanosensation provides animals with important sensory information in addition to olfaction and gustation during feeding behavior. Here, we used Drosophila melanogaster larvae to investigate the role of softness sensing in behavior and learning. In the natural environment, larvae need to dig into soft foods for feeding. Finding foods that are soft enough to dig into is likely to be essential for their survival. We report that larvae can discriminate between different agar concentrations and prefer softer agar. Interestingly, we show that larvae on a harder surface search for a softer surface using memory associated with an odor, and that they evaluate foods by balancing softness and sweetness. These findings suggest that larvae integrate mechanosensory information with chemosensory input while foraging. Moreover, we found that the larval preference for softness is affected by genetic background.
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Drosophila melanogaster/crescimento & desenvolvimento , Comportamento Alimentar , Aprendizagem , Ágar , Animais , Drosophila melanogaster/fisiologia , Larva/fisiologia , Mecanorreceptores , Memória , Olfato , PaladarRESUMO
The coordination of growth with nutritional status is essential for proper development and physiology. Nutritional information is mostly perceived by peripheral organs before being relayed to the brain, which modulates physiological responses. Hormonal signaling ensures this organ-to-organ communication, and the failure of endocrine regulation in humans can cause diseases including obesity and diabetes. In Drosophila melanogaster, the fat body (adipose tissue) has been suggested to play an important role in coupling growth with nutritional status. Here, we show that the peripheral tissue-derived peptide hormone CCHamide-2 (CCHa2) acts as a nutrient-dependent regulator of Drosophila insulin-like peptides (Dilps). A BAC-based transgenic reporter revealed strong expression of CCHa2 receptor (CCHa2-R) in insulin-producing cells (IPCs) in the brain. Calcium imaging of brain explants and IPC-specific CCHa2-R knockdown demonstrated that peripheral-tissue derived CCHa2 directly activates IPCs. Interestingly, genetic disruption of either CCHa2 or CCHa2-R caused almost identical defects in larval growth and developmental timing. Consistent with these phenotypes, the expression of dilp5, and the release of both Dilp2 and Dilp5, were severely reduced. Furthermore, transcription of CCHa2 is altered in response to nutritional levels, particularly of glucose. These findings demonstrate that CCHa2 and CCHa2-R form a direct link between peripheral tissues and the brain, and that this pathway is essential for the coordination of systemic growth with nutritional availability. A mammalian homologue of CCHa2-R, Bombesin receptor subtype-3 (Brs3), is an orphan receptor that is expressed in the islet ß-cells; however, the role of Brs3 in insulin regulation remains elusive. Our genetic approach in Drosophila melanogaster provides the first evidence, to our knowledge, that bombesin receptor signaling with its endogenous ligand promotes insulin production.
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Proteínas de Drosophila/biossíntese , Proteínas de Drosophila/genética , Insulina/metabolismo , Insulinas/biossíntese , Neuropeptídeos/genética , Receptores da Bombesina/genética , Receptores Odorantes/genética , Animais , Animais Geneticamente Modificados , Encéfalo/metabolismo , Drosophila melanogaster , Corpo Adiposo/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Células Secretoras de Insulina/metabolismo , Insulinas/genética , Neuropeptídeos/biossíntese , Receptores Odorantes/biossínteseRESUMO
The neural substrates that the fruitfly Drosophila uses to sense smell, taste and light share marked structural and functional similarities with ours, providing attractive models to dissect sensory stimulus processing. Here we focus on two of the remaining and less understood prime sensory modalities: graviception and hearing. We show that the fly has implemented both sensory modalities into a single system, Johnston's organ, which houses specialized clusters of mechanosensory neurons, each of which monitors specific movements of the antenna. Gravity- and sound-sensitive neurons differ in their response characteristics, and only the latter express the candidate mechanotransducer channel NompC. The two neural subsets also differ in their central projections, feeding into neural pathways that are reminiscent of the vestibular and auditory pathways in our brain. By establishing the Drosophila counterparts of these sensory systems, our findings provide the basis for a systematic functional and molecular dissection of how different mechanosensory stimuli are detected and processed.
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Drosophila melanogaster/fisiologia , Sensação Gravitacional/fisiologia , Audição/fisiologia , Células Receptoras Sensoriais/fisiologia , Animais , Sinalização do Cálcio , Proteínas de Drosophila/genética , Drosophila melanogaster/anatomia & histologia , Drosophila melanogaster/metabolismo , Regulação da Expressão Gênica , Canais Iônicos/genética , Células Receptoras Sensoriais/metabolismo , Transdução de Sinais , Canais de Potencial de Receptor Transitório , VibraçãoRESUMO
Behavioural responses to wind are thought to have a critical role in controlling the dispersal and population genetics of wild Drosophila species, as well as their navigation in flight, but their underlying neurobiological basis is unknown. We show that Drosophila melanogaster, like wild-caught Drosophila strains, exhibits robust wind-induced suppression of locomotion in response to air currents delivered at speeds normally encountered in nature. Here we identify wind-sensitive neurons in Johnston's organ, an antennal mechanosensory structure previously implicated in near-field sound detection (reviewed in refs 5 and 6). Using enhancer trap lines targeted to different subsets of Johnston's organ neurons, and a genetically encoded calcium indicator, we show that wind and near-field sound (courtship song) activate distinct populations of Johnston's organ neurons, which project to different regions of the antennal and mechanosensory motor centre in the central brain. Selective genetic ablation of wind-sensitive Johnston's organ neurons in the antenna abolishes wind-induced suppression of locomotion behaviour, without impairing hearing. Moreover, different neuronal subsets within the wind-sensitive population respond to different directions of arista deflection caused by air flow and project to different regions of the antennal and mechanosensory motor centre, providing a rudimentary map of wind direction in the brain. Importantly, sound- and wind-sensitive Johnston's organ neurons exhibit different intrinsic response properties: the former are phasically activated by small, bi-directional, displacements of the aristae, whereas the latter are tonically activated by unidirectional, static deflections of larger magnitude. These different intrinsic properties are well suited to the detection of oscillatory pulses of near-field sound and laminar air flow, respectively. These data identify wind-sensitive neurons in Johnston's organ, a structure that has been primarily associated with hearing, and reveal how the brain can distinguish different types of air particle movements using a common sensory organ.
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Movimentos do Ar , Percepção Auditiva/fisiologia , Drosophila melanogaster/fisiologia , Células Receptoras Sensoriais/fisiologia , Animais , Comportamento Animal/fisiologia , Fenômenos Eletrofisiológicos/fisiologia , Mecanorreceptores/fisiologiaRESUMO
As observed in human language learning and song learning in birds, the fruit fly Drosophila melanogaster changes its auditory behaviors according to prior sound experiences. This phenomenon, known as song preference learning in flies, requires GABAergic input to pC1 neurons in the brain, with these neurons playing a key role in mating behavior. The neural circuit basis of this GABAergic input, however, is not known. Here, we find that GABAergic neurons expressing the sex-determination gene doublesex are necessary for song preference learning. In the brain, only four doublesex-expressing GABAergic neurons exist per hemibrain, identified as pCd-2 neurons. pCd-2 neurons directly, and in many cases mutually, connect with pC1 neurons, suggesting the existence of reciprocal circuits between them. Moreover, GABAergic and dopaminergic inputs to doublesex-expressing GABAergic neurons are necessary for song preference learning. Together, this study provides a neural circuit model that underlies experience-dependent auditory plasticity at a single-cell resolution.
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Acoustic communication plays an important role during the courtship of many mosquito species. Male mosquitoes show strong attraction to female wing beat frequencies, mediated via spectral matching between female wing beat frequency and male ear mechanical tuning frequency. Such acoustic communication typically occurs within swarms, male-dominated aggregations with species-specific properties. Despite hundreds of relevant publications being available, the lack of a central platform hosting all associated data hinders research efforts and limits cross-species comparisons. Here, we introduce MACSFeD (Mosquito Acoustic Communication and Swarming Features Database), an interactive platform for the exploration of our comprehensive database containing 251 unique reports focusing on different aspects of mosquito acoustic communication, including hearing function, wing beat frequency and phonotaxis, as well as male swarming parameters. MACSFeD serves as an easily accessible, efficient, and robust data visualization tool for mosquito acoustic communication research. We envision that further in-depth studies could arise following the use of this new platform. Database URL: https://minmatt.shinyapps.io/MACSFeD/.
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Acústica , Culicidae , Bases de Dados Factuais , Animais , Culicidae/fisiologia , Masculino , Feminino , Comunicação AnimalRESUMO
When Aedes albopictus mosquitoes invade regions predominated by Aedes aegypti, either the latter can be displaced or the species can coexist, with potential consequences on disease transmission. Males from both species identify females by listening for her flight sounds. Comparing male hearing systems may provide insight into how hearing could prevent interspecific mating. Here, we show that species-specific differences in female wing beat frequencies are reflected in differences in male ear mechanical tuning frequencies and sound response profiles. Though Aedes albopictus males are attracted to sound, they do not readily display abdominal bending, unlike Aedes aegypti. We observed interspecific differences in male ear mechanical, but not electrical, tuning, suggesting a conserved primary auditory processing pathway. Our work suggests a potential role for hearing in the premating isolation of Aedes aegypti and Aedes albopictus, with implications for predicting future dynamics in their sympatric relationships and our understanding of mosquito acoustic communication.
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Modification of guanosine to N7-methylguanosine (m7G) in the variable loop region of tRNA is catalyzed by the METTL1/WDR4 heterodimer and stabilizes target tRNA. Here, we reveal essential functions of Mettl1 in Drosophila fertility. Knockout of Mettl1 (Mettl1-KO) causes no major effect on the development of non-gonadal tissues, but abolishes the production of elongated spermatids and mature sperm, which is fully rescued by expression of a Mettl1-transgene, but not a catalytic-dead Mettl1 transgene. This demonstrates that Mettl1-dependent m7G is required for spermatogenesis. Mettl1-KO results in a loss of m7G modification on a subset of tRNAs and decreased tRNA abundance. Ribosome profiling shows that Mettl1-KO led to ribosomes stalling at codons decoded by tRNAs that were reduced in abundance. Mettl1-KO also significantly reduces the translation efficiency of genes involved in elongated spermatid formation and sperm stability. Germ cell-specific expression of Mettl1 rescues disrupted m7G tRNA modification and tRNA abundance in Mettl1-KO testes but not in non-gonadal tissues. Ribosome stalling is much less detectable in non-gonadal tissues than in Mettl1-KO testes. These findings reveal a developmental role for m7G tRNA modification and indicate that m7G modification-dependent tRNA abundance differs among tissues.
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Proteínas de Drosophila , Drosophila melanogaster , Fertilidade , RNA de Transferência , Espermatogênese , Animais , Espermatogênese/genética , Masculino , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , RNA de Transferência/metabolismo , RNA de Transferência/genética , Fertilidade/genética , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Guanosina/metabolismo , Guanosina/análogos & derivados , Metiltransferases/metabolismo , Metiltransferases/genética , Espermatozoides/metabolismo , Ribossomos/metabolismo , Espermátides/metabolismo , Testículo/metabolismo , Técnicas de Inativação de GenesRESUMO
The fruit fly Drosophila melanogaster responds behaviorally to sound, gravity, and wind. Exposure to male courtship songs results in reduced locomotion in females, whereas males begin to chase each other. When agitated, fruit flies tend to move against gravity. When faced with air currents, they 'freeze' in place. Based on recent studies, Johnston's hearing organ, the antennal ear of the fruit fly, serves as a sensor for all of these mechanosensory stimuli. Compartmentalization of sense cells in Johnston's organ into vibration-sensitive and deflection-sensitive neural groups allows this single organ to mediate such varied functions. Sound and gravity/wind signals sensed by these two neuronal groups travel in parallel from the fly ear to the brain, feeding into neural pathways reminiscent of the auditory and vestibular pathways in the human brain. Studies of the similarities between mammals and flies will lead to a better understanding of the principles of how sound and gravity information is encoded in the brain. Here, we review recent advances in our understanding of these principles and discuss the advantages of the fruit fly as a model system to explore the fundamental principles of how neural circuits and their ensembles process and integrate sensory information in the brain.
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Antenas de Artrópodes/inervação , Percepção Auditiva , Drosophila melanogaster/fisiologia , Gravitação , Sensação Gravitacional , Mecanotransdução Celular , Som , Vento , Animais , Vias Auditivas/fisiologia , Comportamento Animal , Voo Animal , Mecanorreceptores/fisiologia , VibraçãoRESUMO
In internal fertilization animals, maintaining a copulation posture facilitates the process of transporting gametes from male to female. Here, we present a protocol to investigate the neural basis for copulation posture of fruit flies using a closed-loop real-time optogenetic system. We describe steps for using deep learning analysis to enable optogenetic manipulation of neural activity only during copulation with high efficiency. This system can be applied to various animal behaviors other than copulation. For complete details on the use and execution of this protocol, please refer to Yamanouchi et al. (2023).1.
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Copulação , Drosophila , Animais , Masculino , Feminino , Optogenética/métodos , Comportamento Animal , Sistemas ComputacionaisRESUMO
In internal fertilization animals, reproductive success depends on maintaining copulation until gametes are transported from male to female. In Drosophila melanogaster, mechanosensation in males likely contributes to copulation maintenance, but its molecular underpinning remains to be identified. Here we show that the mechanosensory gene piezo and its' expressing neurons are responsible for copulation maintenance. An RNA-seq database search and subsequent mutant analysis revealed the importance of piezo for maintaining male copulation posture. piezo-GAL4-positive signals were found in the sensory neurons of male genitalia bristles, and optogenetic inhibition of piezo-expressing neurons in the posterior side of the male body during copulation destabilized posture and terminated copulation. Our findings suggest that the mechanosensory system of male genitalia through Piezo channels plays a key role in copulation maintenance and indicate that Piezo may increase male fitness during copulation in flies.
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Acoustic communication signals diversify even on short evolutionary time scales. To understand how the auditory system underlying acoustic communication could evolve, we conducted a systematic comparison of the early stages of the auditory neural circuit involved in song information processing between closely-related fruit-fly species. Male Drosophila melanogaster and D. simulans produce different sound signals during mating rituals, known as courtship songs. Female flies from these species selectively increase their receptivity when they hear songs with conspecific temporal patterns. Here, we firstly confirmed interspecific differences in temporal pattern preferences; D. simulans preferred pulse songs with longer intervals than D. melanogaster. Primary and secondary song-relay neurons, JO neurons and AMMC-B1 neurons, shared similar morphology and neurotransmitters between species. The temporal pattern preferences of AMMC-B1 neurons were also relatively similar between species, with slight but significant differences in their band-pass properties. Although the shift direction of the response property matched that of the behavior, these differences are not large enough to explain behavioral differences in song preferences. This study enhances our understanding of the conservation and diversification of the architecture of the early-stage neural circuit which processes acoustic communication signals.
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Drosophila melanogaster , Drosophila , Animais , Masculino , Feminino , Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Corte , Evolução Biológica , Neurônios , Drosophila simulans , Comportamento Sexual Animal/fisiologia , Vocalização Animal/fisiologiaRESUMO
Lactic acid bacteria (LAB) influence multiple aspects of host brain function via the production of active metabolites in the gut, which is known as the pre/probiotic action. However, little is known about the biogenic effects of LAB on host brain function. Here, we reported that the Lactobacillus plantarum SBT2227 promoted sleep in Drosophila melanogaster. Administration of SBT2227 primarily increased the amount of sleep and decreased sleep latency at the beginning of night-time. The sleep-promoting effects of SBT2227 were independent of the existing gut flora. Furthermore, heat treatment or mechanical crushing of SBT2227 did not suppress the sleep-promoting effects, indicative of biogenic action. Transcriptome analysis and RNAi mini-screening for gut-derived peptide hormones revealed the requirement of neuropeptide F, a homolog of the mammalian neuropeptide Y, for the action of SBT2227. These biogenic effects of SBT2227 on the host sleep provide new insights into the interaction between the brain and gut bacteria.
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Many animal species form groups. Group characteristics differ between species, suggesting that the decision-making of individuals for grouping varies across species. However, the actual decision-making properties that lead to interspecific differences in group characteristics remain unclear. Here, we compared the group formation processes of two Drosophilinae fly species, Colocasiomyia alocasiae and Drosophila melanogaster, which form dense and sparse groups, respectively. A high-throughput tracking system revealed that C. alocasiae flies formed groups faster than D. melanogaster flies, and the probability of C. alocasiae remaining in groups was far higher than that of D. melanogaster. C. alocasiae flies joined groups even when the group size was small, whereas D. melanogaster flies joined groups only when the group size was sufficiently large. C. alocasiae flies attenuated their walking speed when the inter-individual distance between flies became small, whereas such behavioural properties were not clearly observed in D. melanogaster. Furthermore, depriving C. alocasiae flies of visual input affected grouping behaviours, resulting in a severe reduction in group formation. These findings show that C. alocasiae decision-making regarding grouping, which greatly depends on vision, is significantly different from D. melanogaster, leading to species-specific group formation properties.
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Male Aedes aegypti (Ae. aegypti) mosquitoes rely on hearing to identify conspecific females for mating, with the male attraction to the sound of flying females ("phonotaxis") an important behavior in the initial courtship stage. Hearing thus represents a promising target for novel methods of mosquito control, and hearing behaviors (such as male phonotaxis) can be targeted via the use of sound traps. These traps unfortunately have proven to be relatively ineffective during field deployment. Shifting the target from hearing behavior to hearing function could therefore offer a novel method of interfering with Ae. aegypti mating. Numerous neurotransmitters, including serotonin (5-hydroxytryptamine, or 5-HT) and octopamine, are expressed in the male ear, with modulation of the latter proven to influence the mechanical responses of the ear to sound. The effect of serotonin modulation however remains underexplored despite its significant role in determining many key behaviors and biological processes of animals. Here we investigated the influence of serotonin on the Ae. aegypti hearing function and behaviors. Using immunohistochemistry, we found significant expression of serotonin in the male and female Ae. aegypti ears. In the male ear, presynaptic sites identified via antibody labelling showed only partial overlap with serotonin. Next, we used RT-qPCR to identify and quantify the expression levels of three different serotonin receptor families (5-HT1, 5-HT2, and 5-HT7) in the mosquito heads and ears. Although all receptors were identified in the ears of both sexes, those from the 5-HT7 family were significantly more expressed in the ears relative to the heads. We then thoracically injected serotonin-related compounds into the mosquitoes and found a significant, reversible effect of serotonin exposure on the male ear mechanical tuning frequency. Finally, oral administration of a serotonin-synthesis inhibitor altered male phonotaxis. The mosquito serotonergic system and its receptors thus represent interesting targets for novel methods of mosquito, and thus disease, control.