RESUMO
Coordinated activity across networks of neurons is a hallmark of both resting and active behavioural states in many species1-5. These global patterns alter energy metabolism over seconds to hours, which underpins the widespread use of oxygen consumption and glucose uptake as proxies of neural activity6,7. However, whether changes in neural activity are causally related to metabolic flux in intact circuits on the timescales associated with behaviour is unclear. Here we combine two-photon microscopy of the fly brain with sensors that enable the simultaneous measurement of neural activity and metabolic flux, across both resting and active behavioural states. We demonstrate that neural activity drives changes in metabolic flux, creating a tight coupling between these signals that can be measured across brain networks. Using local optogenetic perturbation, we demonstrate that even transient increases in neural activity result in rapid and persistent increases in cytosolic ATP, which suggests that neuronal metabolism predictively allocates resources to anticipate the energy demands of future activity. Finally, our studies reveal that the initiation of even minimal behavioural movements causes large-scale changes in the pattern of neural activity and energy metabolism, which reveals a widespread engagement of the brain. As the relationship between neural activity and energy metabolism is probably evolutionarily ancient and highly conserved, our studies provide a critical foundation for using metabolic proxies to capture changes in neural activity.
Assuntos
Comportamento Animal , Encéfalo/citologia , Encéfalo/fisiologia , Drosophila melanogaster/metabolismo , Drosophila melanogaster/fisiologia , Redes e Vias Metabólicas , Neurônios/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Encéfalo/metabolismo , Drosophila melanogaster/citologia , Metabolismo Energético , Feminino , Masculino , Vias Neurais , Optogenética , DescansoRESUMO
Anatomical connectivity can constrain both a neural circuit's function and its underlying computation. This principle has been demonstrated for many small, defined neural circuits. For example, connectome reconstructions have informed models for direction selectivity in the vertebrate retina1,2 as well as the Drosophila visual system.3 In these cases, the circuit in question is relatively compact, well-defined, and has known functions. However, how the connectome constrains global properties of large-scale networks, across multiple brain regions or the entire brain, is incompletely understood. As the availability of partial or complete connectomes expands to more systems and species4-8 it becomes critical to understand how this detailed anatomical information can inform our understanding of large-scale circuit function.9,10 Here, we use data from the Drosophila connectome4 in conjunction with whole-brain in vivo imaging11 to relate structural and functional connectivity in the central brain. We find a strong relationship between resting-state functional correlations and direct region-to-region structural connectivity. We find that the relationship between structure and function varies across the brain, with some regions displaying a tight correspondence between structural and functional connectivity whereas others, including the mushroom body, are more strongly dependent on indirect connections. Throughout this work, we observe features of structural and functional networks in Drosophila that are strikingly similar to those seen in mammalian cortex, including in the human brain. Given the vast anatomical and functional differences between Drosophila and mammalian nervous systems, these observations suggest general principles that govern brain structure, function, and the relationship between the two.
Assuntos
Conectoma , Animais , Encéfalo , Córtex Cerebral , Drosophila , Humanos , Imageamento por Ressonância Magnética , Rede NervosaRESUMO
A long-standing goal of neuroscience has been to understand how computations are implemented across large-scale brain networks. By correlating spontaneous activity during "resting states" [1], studies of intrinsic brain networks in humans have demonstrated a correspondence with task-related activation patterns [2], relationships to behavior [3], and alterations in processes such as aging [4] and brain disorders [5], highlighting the importance of resting-state measurements for understanding brain function. Here, we develop methods to measure intrinsic functional connectivity in Drosophila, a powerful model for the study of neural computation. Recent studies using calcium imaging have measured neural activity at high spatial and temporal resolution in zebrafish, Drosophila larvae, and worms [6-10]. For example, calcium imaging in the zebrafish brain recently revealed correlations between the midbrain and hindbrain, demonstrating the utility of measuring intrinsic functional connections in model organisms [8]. An important component of human connectivity research is the use of brain atlases to compare findings across individuals and studies [11]. An anatomical atlas of the central adult fly brain was recently described [12]; however, combining an atlas with whole-brain calcium imaging has yet to be performed in vivo in adult Drosophila. Here, we measure intrinsic functional connectivity in Drosophila by acquiring calcium signals from the central brain. We develop an alignment procedure to assign functional data to atlas regions and correlate activity between regions to generate brain networks. This work reveals a large-scale architecture for neural communication and provides a framework for using Drosophila to study functional brain networks.
Assuntos
Mapeamento Encefálico/métodos , Cálcio/metabolismo , Drosophila melanogaster/fisiologia , Animais , Encéfalo/fisiologia , FemininoRESUMO
Classifying sensory experiences as either novel or familiar represents a fundamental challenge to neural processing. In this issue of Cell, Hattori et al. describe a circuit mechanism by which a novel stimulus that initially interests a fruit fly turns into a familiar one.
Assuntos
Reconhecimento Psicológico , HumanosRESUMO
Nutrient deprivation can lead to dramatic changes in feeding behavior, including acceptance of foods that are normally rejected. In flies, this behavioral shift depends in part on reciprocal sensitization and desensitization of sweet and bitter taste, respectively. However, the mechanisms for bitter taste modulation remain unclear. Here, we identify a set of octopaminergic/tyraminergic neurons, named OA-VLs, that directly modulate bitter sensory neuron output in response to starvation. OA-VLs are in close proximity to bitter sensory neuron axon terminals and show reduced tonic firing following starvation. We find that octopamine and tyramine potentiate bitter sensory neuron responses, suggesting that starvation-induced reduction in OA-VL activity depotentiates bitter taste. Consistent with this model, artificial silencing of OA-VL activity induces a starvation-like reduction in bitter sensory neuron output. These results demonstrate that OA-VLs mediate a critical step in starvation-dependent bitter taste modulation, allowing flies to dynamically balance the risks associated with bitter food consumption against the threat of severe starvation.
Assuntos
Drosophila melanogaster/fisiologia , Privação de Alimentos , Depressão Sináptica de Longo Prazo , Percepção Gustatória , Animais , Feminino , Células Receptoras Sensoriais/fisiologiaRESUMO
Hunger and thirst are ancient homeostatic drives for food and water consumption. Although molecular and neural mechanisms underlying these drives are currently being uncovered, less is known about how hunger and thirst interact. Here, we use molecular genetic, behavioral, and anatomical studies in Drosophila to identify four neurons that modulate food and water consumption. Activation of these neurons promotes sugar consumption and restricts water consumption, whereas inactivation promotes water consumption and restricts sugar consumption. By calcium imaging studies, we show that these neurons are directly regulated by a hormone signal of nutrient levels and by osmolality. Finally, we identify a hormone receptor and an osmolality-sensitive ion channel that underlie this regulation. Thus, a small population of neurons senses internal signals of nutrient and water availability to balance sugar and water consumption. Our results suggest an elegant mechanism by which interoceptive neurons oppositely regulate homeostatic drives to eat and drink.
Assuntos
Neurônios/metabolismo , Transdução de Sinais , Animais , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Homeostase , Fome , Concentração Osmolar , Receptores de Glucagon/metabolismo , Sacarose/metabolismo , Sede , Canais de Potencial de Receptor Transitório/metabolismoRESUMO
Linking structural changes in neurons to animal behavior has proven challenging. New findings by Pesakou et al. tie daily cycles of axon arbor extension and retraction, mediated by Rho activity, to circadian and seasonal patterns of behavior in the fruit fly.
Assuntos
Ritmo Circadiano , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Proteínas rho de Ligação ao GTP/metabolismo , AnimaisRESUMO
The sense of taste is critical in determining the nutritional suitability of foods. Sweet and bitter are primary taste modalities in mammals, and their behavioral relevance is similar in flies. Sweet taste drives the appetitive response to energy sources, whereas bitter taste drives avoidance of potential toxins and also suppresses the sweet response [1, 2]. Despite their importance to survival, little is known about the neural circuit mechanisms underlying integration of sweet and bitter taste. Here, we describe a presynaptic gain control mechanism in Drosophila that differentially affects sweet and bitter taste channels and mediates integration of these opposing stimuli. Gain control is known to play an important role in fly olfaction, where GABAB receptor (GABABR) mediates intra- and interglomerular presynaptic inhibition of sensory neuron output [3-5]. In the taste system, we find that gustatory receptor neurons (GRNs) responding to sweet compounds express GABABR, whereas those that respond to bitter do not. GABABR mediates presynaptic inhibition of calcium responses in sweet GRNs, and both sweet and bitter stimuli evoke GABAergic neuron activity in the vicinity of GRN axon terminals. Pharmacological blockade and genetic reduction of GABABR both lead to increased sugar responses and decreased suppression of the sweet response by bitter compounds. We propose a model in which GABA acts via GABABR to expand the dynamic range of sweet GRNs through presynaptic gain control and suppress the output of sweet GRNs in the presence of opposing bitter stimuli.
Assuntos
Drosophila melanogaster/fisiologia , Percepção Gustatória , Animais , Feminino , Células Receptoras Sensoriais/fisiologia , Sacarose/químicaRESUMO
Feeding is dynamically regulated by the palatability of the food source and the physiological needs of the animal. How consumption is controlled by external sensory cues and internal metabolic state remains under intense investigation. Here, we identify four GABAergic interneurons in the Drosophila brain that establish a central feeding threshold which is required to inhibit consumption. Inactivation of these cells results in indiscriminate and excessive intake of all compounds, independent of taste quality or nutritional state. Conversely, acute activation of these neurons suppresses consumption of water and nutrients. The output from these neurons is required to gate activity in motor neurons that control meal initiation and consumption. Thus, our study reveals a layer of inhibitory control in feeding circuits that is required to suppress a latent state of unrestricted and nonselective consumption.
Assuntos
Comportamento Alimentar/fisiologia , Neurônios GABAérgicos/fisiologia , Interneurônios/fisiologia , Animais , Animais Geneticamente Modificados , Drosophila , Feminino , Trato Gastrointestinal/inervação , Trato Gastrointestinal/fisiologiaRESUMO
The decision to engage in one behavior often precludes the selection of others, suggesting cross-inhibition between incompatible behaviors. For example, the likelihood to initiate feeding might be influenced by an animal's commitment to other behaviors. Here, we examine the modulation of feeding behavior in the fruit fly, Drosophila melanogaster, and identify a pair of interneurons in the ventral nerve cord that is activated by stimulation of mechanosensory neurons and inhibits feeding initiation, suggesting that these neurons suppress feeding while the fly is walking. Conversely, inhibiting activity in these neurons promotes feeding initiation and inhibits locomotion. These studies demonstrate the mutual exclusivity between locomotion and feeding initiation in the fly, isolate interneurons that influence this behavioral choice, and provide a framework for studying the neural basis for behavioral exclusivity in Drosophila.
Assuntos
Comportamento de Escolha/fisiologia , Comportamento Alimentar/fisiologia , Interneurônios/fisiologia , Locomoção/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/genética , Análise de Variância , Animais , Animais Geneticamente Modificados , Encéfalo/citologia , Calmodulina/genética , Calmodulina/metabolismo , Relação Dose-Resposta a Droga , Drosophila , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Comportamento Alimentar/efeitos dos fármacos , Privação de Alimentos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Interneurônios/classificação , Locomoção/efeitos dos fármacos , Quinase de Cadeia Leve de Miosina/genética , Quinase de Cadeia Leve de Miosina/metabolismo , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Rodopsina/genética , Sacarose/administração & dosagem , Edulcorantes/administração & dosagem , Temperatura , Proteínas Virais de Fusão/genética , Proteínas Virais de Fusão/metabolismoRESUMO
For an animal to survive in a constantly changing environment, its behavior must be shaped by the complex milieu of sensory stimuli it detects, its previous experience, and its internal state. Although taste behaviors in the fly are relatively simple, with sugars eliciting acceptance behavior and bitter compounds avoidance, these behaviors are also plastic and are modified by intrinsic and extrinsic cues, such as hunger and sensory stimuli. Here, we show that dopamine modulates a simple taste behavior, proboscis extension to sucrose. Conditional silencing of dopaminergic neurons reduces proboscis extension probability, and increased activation of dopaminergic neurons increases extension to sucrose, but not to bitter compounds or water. One dopaminergic neuron with extensive branching in the primary taste relay, the subesophageal ganglion, triggers proboscis extension, and its activity is altered by satiety state. These studies demonstrate the marked specificity of dopamine signaling and provide a foundation to examine neural mechanisms of feeding modulation in the fly.
Assuntos
Dopamina/farmacologia , Comportamento Alimentar/fisiologia , Preferências Alimentares/fisiologia , Sacarose/administração & dosagem , Edulcorantes/administração & dosagem , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/genética , Animais , Animais Geneticamente Modificados , Antígenos CD8/genética , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/fisiologia , Relação Dose-Resposta a Droga , Drosophila , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Feminino , Privação de Alimentos/fisiologia , Gânglios dos Invertebrados/citologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Canais Iônicos , Canais de Potássio Corretores do Fluxo de Internalização/genética , Canais de Potássio Corretores do Fluxo de Internalização/metabolismo , Probabilidade , Proteínas R-SNARE/genética , Proteínas R-SNARE/metabolismo , Resposta de Saciedade/efeitos dos fármacos , Resposta de Saciedade/fisiologia , Canal de Cátion TRPA1 , Canais de Cátion TRPC/genética , Canais de Cátion TRPC/metabolismo , Fatores de Transcrição/genética , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
In insects, increasing evidence suggests that small secreted pheromone binding proteins (PBPs) and odorant binding proteins (OBPs) are important for normal olfactory detection of airborne pheromones and odorants far from their source. In contrast, it is unknown whether extracellular ligand binding proteins participate in perception of less volatile chemicals, including many pheromones, that are detected by direct contact with chemosensory organs. CheB42a, a small Drosophila melanogaster protein unrelated to known PBPs or OBPs, is expressed and likely secreted in only a small subset of gustatory sensilla on males' front legs, the site of gustatory perception of contact pheromones. Here we show that CheB42a is expressed specifically in the sheath cells surrounding the taste neurons expressing Gr68a, a putative gustatory pheromone receptor for female cuticular hydrocarbons that stimulate male courtship. Surprisingly, however, CheB42a mutant males attempt to copulate with females earlier and more frequently than control males. Furthermore, CheB42a mutant males also attempt to copulate more frequently with other males that secrete female-specific cuticular hydrocarbon pheromones, but not with females lacking cuticular hydrocarbons. Together, these data indicate that CheB42a is required for a normal gustatory response to female cuticular hydrocarbon pheromones that modulate male courtship.
Assuntos
Copulação/fisiologia , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Atrativos Sexuais/metabolismo , Animais , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Extremidades/anatomia & histologia , Extremidades/fisiologia , Feminino , Proteínas de Fluorescência Verde/análise , Hidrocarbonetos/metabolismo , MasculinoRESUMO
Odorants and pheromones as well as sweet- and bitter-tasting small molecules are perceived through activation of G protein-coupled chemosensory receptors. In contrast, gustatory detection of salty and sour tastes may involve direct gating of sodium channels of the DEG/ENaC family by sodium and hydrogen ions, respectively. We have found that ppk25, a Drosophila melanogaster gene encoding a DEG/ENaC channel subunit, is expressed at highest levels in the male appendages responsible for gustatory and olfactory detection of female pheromones: the legs, wings, and antennae. Mutations in the ppk25 gene reduce or even abolish male courtship response to females in the dark, conditions under which detection of female pheromones is an essential courtship-activating sensory input. In contrast, the same mutations have no effect on other behaviors tested. Importantly, ppk25 mutant males that show no response to females in the dark execute all of the normal steps of courtship behavior in the presence of visible light, suggesting that ppk25 is required for activation of courtship behavior by chemosensory perception of female pheromones. Finally, a ppk25 mutant allele predicted to encode a truncated protein has dominant-negative properties, suggesting that the normal Ppk25 protein acts as part of a multiprotein complex. Together, these results indicate that ppk25 is necessary for response to female pheromones by D. melanogaster males, and suggest that members of the DEG/ENaC family of genes play a wider role in chemical senses than previously suspected.
Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/metabolismo , Feromônios/farmacologia , Canais de Sódio/metabolismo , Envelhecimento/fisiologia , Alelos , Animais , Corte , Elementos de DNA Transponíveis/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Feminino , Regulação da Expressão Gênica/genética , Íntrons/genética , Masculino , Mutagênese Insercional , Especificidade de Órgãos , Feromônios/metabolismo , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Canais de Sódio/genéticaRESUMO
Control of nitric oxide (NO) synthesis in the constitutive nitric-oxide synthases (NOS) by calcium/calmodulin is exerted through the regulation of electron transfer from NADPH through the reductase domains. This process has been shown previously to involve the calmodulin binding site, the autoinhibitory insertion in the FMN binding domain, and the C-terminal tail. Smaller sequence elements also appear to correlate with control. Although some of these elements appear well positioned to function in control, they are poorly conserved; their role in control is neither well established nor defined by available information. In this study mutations have been induced in the small insertion of the hinge subdomain, which has been shown recently to form a beta hairpin in structural studies of the neuronal NOS reductase domains adjacent to the calmodulin site and the autoinhibitory element. Modification of the small insertion in neuronal NOS tends to increase cytochrome c reduction but not NO synthetic activity; some modifications or deletions in the corresponding region in endothelial NOS modestly increase activity under some conditions. Unexpectedly, some minor changes in the sequence introduce a loss in the content of heme relative to flavin cofactors. Taken together, these results suggest that the small insertion protects the calmodulin binding site and that it may be a modulator of NOS activity.