Crossmodal visual input for odor tracking during fly flight

Flies generate robust and high-performance olfactory and visual behaviors. Adult fruit flies can distinguish small differences in odor concentration across antennae separated by less than 1 mm [1], and a single olfactory sensory neuron is sufficient for near-normal gradient tracking in larvae [2]. During flight a male housefly chasing a female executes a corrective turn within 40 ms after a course deviation by its target [3]. The challenges imposed by flying apparently benefit from the tight integration of unimodal sensory cues. Crossmodal interactions reduce the discrimination threshold for unimodal memory retrieval by enhancing stimulus salience [4], and dynamic crossmodal processing is required for odor search during free flight because animals fail to locate an odor source in the absence of rich visual feedback [5]. The visual requirements for odor localization are unknown. We tethered a hungry fly in a magnetic field, allowing it to yaw freely, presented odor plumes, and examined how visual cues influence odor tracking. We show that flies are unable to use a small-field object or landmark to assist plume tracking, whereas odor activates wide-field optomotor course control to enable accurate orientation toward an attractive food odor.     

A Single-fly Assay for Foraging Behavior in Drosophila

For many animals, hunger promotes changes in the olfactory system in a manner that facilitates the search for appropriate food sources. In this video article, we describe an automated assay to measure the effect of hunger or satiety on olfactory dependent food search behavior in the adult fruit fly Drosophila melanogaster. In a light-tight box illuminated by red light that is invisible to fruit flies, a camera linked to custom data acquisition software monitors the position of six flies simultaneously. Each fly is confined to walk in individual arenas containing a food odor at the center. The testing arenas rest on a porous floor that functions to prevent odor accumulation. Latency to locate the odor source, a metric that reflects olfactory sensitivity under different physiological states, is determined by software analysis. Here, we discuss the critical mechanics of running this behavioral paradigm and cover specific issues regarding fly loading, odor contamination, assay temperature, data quality, and statistical analysis.     

A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila

It has been clear for many years that insects use visual cues to stabilize their heading in a wind stream. Many animals track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Also, the recent deluge of research on the neurophysiology and neurobehavioral genetics of olfaction in Drosophila has motivated ever more technically sophisticated and quantitative behavioral assays. Here, we modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. A fly is glued to a small steel pin and suspended in a magnetic field that enables it to yaw freely. Small diameter food odor plumes are directed downward over the fly s head, eliciting stable tracking by a hungry fly. Here we focus on the critical mechanics of tethering, aligning the magnets, devising the odor plume, and confirming stable odor tracking.Open in a separate windowClick here to view.^{(57M, flv)}     

High-resolution Quantification of Odor-guided Behavior in Drosophila melanogaster Using the Flywalk Paradigm

In their natural environment, insects such as the vinegar fly Drosophila melanogaster are bombarded with a huge amount of chemically distinct odorants. To complicate matters even further, the odors detected by the insect nervous system usually are not single compounds but mixtures whose composition and concentration ratios vary. This leads to an almost infinite amount of different olfactory stimuli which have to be evaluated by the nervous system.To understand which aspects of an odor stimulus determine its evaluation by the fly, it is therefore desirable to efficiently examine odor-guided behavior towards many odorants and odor mixtures. To directly correlate behavior to neuronal activity, behavior should be quantified in a comparable time frame and under identical stimulus conditions as in neurophysiological experiments. However, many currently used olfactory bioassays in Drosophila neuroethology are rather specialized either towards efficiency or towards resolution.Flywalk, an automated odor delivery and tracking system, bridges the gap between efficiency and resolution. It allows the determination of exactly when an odor packet stimulated a freely walking fly, and to determine the animal´s dynamic behavioral reaction.     

视觉和嗅觉信号对果蝇食物搜寻行为的协同作用

为了探索视觉和嗅觉信号在昆虫食物搜寻过程中的作用, 本研究利用杨梅和橘子为引诱物, 在实验室条件下测定了嗅觉和视觉信号诱集到的黑腹果蝇Drosophila melanogaster数量, 分析了嗅觉经历对果蝇嗅觉和视觉食物搜寻的影响。发现同源性嗅觉和视觉信号存在的杨梅诱集到的果蝇数量显著大于单一的视觉信号和嗅觉信号, 但异源性嗅觉和视觉信号组合诱集到的果蝇数量和单独的嗅觉信号相似。嗅觉信号预处理不仅能够显著增加嗅觉信号诱集到的果蝇数量, 其中杨梅嗅觉信号对杨梅预处理果蝇的吸引能力与视觉和嗅觉信号存在的杨梅相似, 而且异源性嗅觉和视觉信号组合诱集到的预处理果蝇数量也不低于视觉和嗅觉信号存在的杨梅。另外杨梅嗅觉信号预处理也能够显著增强杨梅视觉信号诱集到的果蝇数量。但嗅觉预处理并不会改变同源性视觉和嗅觉信号组合诱集到的果蝇数量。本研究表明, 果蝇同时利用视觉和嗅觉信号进行食物搜寻, 因此同源性视觉和嗅觉信号在果蝇诱集过程中具有协同作用。另外果蝇具有较强的记忆和学习能力, 能够将记忆中的嗅觉信号应用于食物搜寻。本研究结果不仅有利于我们了解果蝇在自然状态下的食物搜寻机制, 而且有利于开发更有效的果蝇新型诱捕器。     

Behavioral responses of female oriental fruit flies to the odor of papayas at three ripeness stages in a laboratory flight tunnel (Diptera: Tephritidae)

Behavioral responses of adult female oriental fruit flies, Dacus dorsalisHendel, to the odor of papayas from three ripeness classes were studied using a threechoice flight tunnel bioassay. Laboratoryreared flies were allowed to respond freely to any of three papaya odors (mature green, colorbreak to one-fourth ripe, and one-half to full ripe) emanating from identical (spherical) fruit models. Five behaviors were measured in assessing the fly's relative attraction to the odors (number of landings), arrestment (total fly seconds on sphere), fly-fly interactions on the fruit model (maximum and modal fly density), and acceptance for oviposition (total eggs laid). Females showed no significant difference in total fly landings based on all age classes combined. Significant differences were noted among age classes. Females spent more total time on the sphere and showed a higher maximum density and modal fly density to ripe fruit than to green fruit odors. Ovipositional acceptance of fruit models based on the total number of eggs laid in a sphere was greater in response to the ripefruit odor than to the other two odor classes. Olfactorystimulated behavioral responses of females to the odor of ripe papayas were significantly different from the other ripeness classes for all behaviors at 8 days postemergence and then declined in 11-day-old flies. Behavioral responses were greater during the afternoon than in the morning. Observations of wild oriental fruit flies to papayas in the field indicated a preference for residing on riper fruit. The results of this study are discussed with regard to the role of olfactory inputs generated by the odor of ripening fruit on female attraction and oviposition behavior resulting in infestation of papayas by oriental fruit fly.     

Flying bats use serial sampling to locate odour sources

Olfactory tracking generally sacrifices speed for sensitivity, but some fast-moving animals appear surprisingly efficient at foraging by smell. Here, we analysed the olfactory tracking strategies of flying bats foraging for fruit. Fruit- and nectar-feeding bats use odour cues to find food despite the sensory challenges derived from fast flight speeds and echolocation. We trained Jamaican fruit-eating bats (Artibeus jamaicensis) to locate an odour reward and reconstructed their flight paths in three-dimensional space. Results confirmed that bats relied upon olfactory cues to locate a reward. Flight paths revealed a combination of odour- and memory-guided search strategies. During ‘inspection flights’, bats significantly reduced flight speeds and flew within approximately 6 cm of possible targets to evaluate the presence or absence of the odour cue. This behaviour combined with echolocation explains how bats maximize foraging efficiency while compensating for trade-offs associated with olfactory detection and locomotion.     

Classical Olfactory Conditioning in the Oriental Fruit Fly,Bactrocera dorsalis

The oriental fruit fly, Bactrocera dorsalis, is a serious pest of fruits and vegetables. Methyl eugenol (ME), a male attractant, is used to against this fly by mass trapping. Control effect may be influenced by learning, which could modify the olfactory response of the fly to this attractant. To collect the behavioral evidence, studies on the capability of this fly for olfactory learning are necessary. We investigated olfactory learning in male flies with a classical olfactory conditioning procedure using restrained individuals under laboratory conditions. The acquisition of the proboscis extension reflex was used as the criterion for conditioning. A high conditioned response level was found in oriental fruit flies when an odor was presented in paired association with a sucrose reward but not when the odor and sucrose were presented unpaired. We also found that the conditioning performance was influenced by the odor concentration, intertrial interval, and starvation time. A slight sensitization elicited by imbibing sucrose was observed. These results indicate that oriental fruit flies have a high capacity to form an olfactory memory as a result of classical conditioning.     

Olfactory Deficits in an Alpha-Synuclein Fly Model of Parkinson’s Disease

Parkinson’s disease (PD) is the most common motor neurodegenerative disorder. Olfactory dysfunction is a prevalent feature of PD. It often precedes motor symptoms by several years and is used in assisting PD diagnosis. However, the cellular and molecular bases of olfactory dysfunction in PD are not known. The fruit fly Drosophila melanogaster, expressing human alpha-synuclein protein or its mutant, A30P, captures several hallmarks of PD and has been successfully used to model PD in numerous studies. First, we report olfactory deficits in fly expressing A30P (A30P), showing deficits in two out of three olfactory modalities, tested – olfactory acuity and odor discrimination. The remaining third modality is odor identification/naming. Second, oxidative stress is an important environmental risk factor of PD. We show that oxidative stress exacerbated the two affected olfactory modalities in younger A30P flies. Third, different olfactory receptor neurons are activated differentially by different odors in flies. In a separate experiment, we show that the odor discrimination deficit in A30P flies is general and not restricted to a specific class of chemical structure. Lastly, by restricting A30P expression to dopamine, serotonin or olfactory receptor neurons, we show that A30P expression in dopamine neurons is necessary for development of both acuity and discrimination deficits, while serotonin and olfactory receptor neurons appeared not involved. Our data demonstrate olfactory deficits in a synuclein fly PD model for exploring olfactory pathology and physiology, and for monitoring PD progression and treatment.     

Flower and fruit volatiles assist host‐plant location in the Tomato fruit fly Neoceratitis cyanescens

Mature females of the tomato fruit fly Neoceratitis cyanescens can detect host fruit at a short distance using only visual stimuli, but little is known about the role of airborne volatile cues in the host searching strategy. A series of experiments is conducted in a laboratory wind tunnel, in which the behavioural responses of individual flies to volatiles from Solanaceae host plants (including tomato Lycopersicum esculentum Mill., bug weed Solanum mauritianum Scop. and Turkey berry Solanum torvum Sw.) are observed, according to some environmental (air speed) and physiological (age and mating status of females, time of day) factors. Mature females respond primarily to specific olfactory cues from blends of flowers or host fruit, preferentially unripe fruit for bug weed, as opposed to ripe fruit for Turkey berry or tomato. Males are also highly attracted by the odour of unripe fruit of bug weed. Wind plays a key role, as shown by the proportion of flies that reach the upwind section of the tunnel in the presence of both fruit odour and air flow (66.7%) and in the absence of either fruit odour (13.3%) or wind (36.7%). In response to fruit volatiles carried by wind, flies embark in a ‘plume tracking’ or ‘aim and shoot' flight, consistent with odour‐conditioned anemotaxis. Females respond to host fruit odour regardless of their age, egg load or mating status, and also more consistently in the afternoon, which is their preferential time of day for egg‐laying. Searching behaviour and response to host volatiles in N. cyanescens are discussed in the light of host‐finding and an adaptive strategy.     

Detection of object motion by a fly neuron during simulated flight

Object detection on the basis of relative motion was investigated in the fly at the neuronal level. A representative of the figure detection cells (FD-cells), the FD1b-cell, was characterized with respect to its responses to optic flow which simulated the presence of an object during translatory flight. The figure detection cells reside in the fly's third visual neuropil and are believed to play a central role in mediating object-directed turning behaviour. The dynamical response properties as well as the mean response amplitudes of the FD1b-cell depend on the temporal frequency of object motion and on the presence or absence of background motion. The responses of the FD1b-cell to object motion during simulated translatory flight were compared to behavioural responses of the fly as obtained with identical stimuli in a previous study. The behavioural responses could only partly be explained on the basis of the FD1b-cell's responses. Further processing between the third visual neuropil and the final motor output has to be assumed which involves (1) facilitation of the object-induced responses during translatory background motion at moderate temporal frequencies, and (2) inhibition of the object-induced turning responses during translatory background motion at high temporal frequencies. Accepted: 9 October 1999     

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.Download video file.^{(56M, flv)}     

How locusts separate pattern flow into its rotatory and translatory components (Orthoptera: Acrididae)

The ability of desert locusts,Schistocerca gregaria, to separate pattern flow within the lateral visual fields into its rotatory and translatory components was studied in tethered flight under open-loop conditions. The optomotor turning behavior results from the sum of compensatory steering and upwind/downwind turning induced by the rotatory and translatory component of pattern flow, respectively. Thereby, the analysis of the visual stimulus is supposedly achieved by linear binocular interaction, i.e., by summation and subtraction of the optomotor effectiveness of the pattern flow on either side. Our results indicate that, in addition, locusts take into account the relative contribution of the rotatory and the translatory stimulus component to the sum total of pattern flow. This yields a factor which modifies the gain of the control loop of either of the response components to give a nonlinear response. It results in a weakening of the behavior upon stimuli composed of rotatory and translatory components. We discuss our results as an adaptation by which an animal avoids inappropriate behavior upon ambiguous stimulus situations.     

Smelling on the fly: sensory cues and strategies for olfactory navigation in Drosophila

Navigating toward (or away from) a remote odor source is a challenging problem that requires integrating olfactory information with visual and mechanosensory cues. Drosophila melanogaster is a useful organism for studying the neural mechanisms of these navigation behaviors. There are a wealth of genetic tools in this organism, as well as a history of inventive behavioral experiments. There is also a large and growing literature in Drosophila on the neural coding of olfactory, visual, and mechanosensory stimuli. Here we review recent progress in understanding how these stimulus modalities are encoded in the Drosophila nervous system. We also discuss what strategies a fly might use to navigate in a natural olfactory landscape while making use of all these sources of sensory information. We emphasize that Drosophila are likely to switch between multiple strategies for olfactory navigation, depending on the availability of various sensory cues. Finally, we highlight future research directions that will be important in understanding the neural circuits that underlie these behaviors.     

A Novel Method for Tracking Individuals of Fruit Fly Swarms Flying in a Laboratory Flight Arena

The growing interest in studying social behaviours of swarming fruit flies, Drosophila melanogaster, has heightened the need for developing tools that provide quantitative motion data. To achieve such a goal, multi-camera three-dimensional tracking technology is the key experimental gateway. We have developed a novel tracking system for tracking hundreds of fruit flies flying in a confined cubic flight arena. In addition to the proposed tracking algorithm, this work offers additional contributions in three aspects: body detection, orientation estimation, and data validation. To demonstrate the opportunities that the proposed system offers for generating high-throughput quantitative motion data, we conducted experiments on five experimental configurations. We also performed quantitative analysis on the kinematics and the spatial structure and the motion patterns of fruit fly swarms. We found that there exists an asymptotic distance between fruit flies in swarms as the population density increases. Further, we discovered the evidence for repulsive response when the distance between fruit flies approached the asymptotic distance. Overall, the proposed tracking system presents a powerful method for studying flight behaviours of fruit flies in a three-dimensional environment.     

果蝇嗅觉分子机理研究进展

黑腹果蝇Drosophila melanogaster是生物学研究的重要模式生物,也是探索研究生物体嗅觉奥秘的理想材料。近年来,由于分子生物学技术在神经科学领域的广泛应用,黑腹果蝇嗅觉机理研究取得了许多重大突破, 对气味分子受体及其识别机理、 嗅觉神经电信号的产生和传递、嗅觉信息的加工、编码以及记忆等方面都有了深入的了解。研究表明, 果蝇约1 300个嗅神经元(olfactory receptor neurons, ORNs)共表达62种不同的气味受体蛋白（olfactory receptor proteins, ORs）, 用以检测和识别其所感受的所有化学气味分子。许多OR所识别的气味分子配体已鉴定出来,普通的气味（如水果的气味）由数种不同的OR组合来识别,而信息素（pheromone）分子则由单种特定的OR来检测。气味信息在嗅神经元内转换成神经电信号,嗅觉电信号沿嗅神经元的轴突传递到触角叶, 再经投射神经元（projection neurons, PNs）将信息送至高级中枢如蘑菇体（mushroom body, MB）和侧角（lateral horn, LH）,最终引发行为反应。在黑腹果蝇嗅觉信息传递通路中,某些蛋白如Dock,N-cadherin,Fruitless等起着重要作用,缺失这些蛋白会导致嗅觉异常。本文对这些研究进展作一综述。     

A cyclic nucleotide-gated channel in the brain regulates olfactory modulation through neuropeptide F in fruit fly Drosophila melanogaster

Olfactory sensing and its modulation are important for the insects in recognizing diverse odors from the environment and in making correct decisions to survive. Identifying new genes involved in olfactory modulation and unveiling their mechanisms may lead us to understand decision making processes in the central nervous system. Here, we report a novel olfactory function of the cyclic nucleotide-gated (CNG) channel CG42260 in modulating ab3A olfactory sensory neurons, which specifically respond to food-derived odors in fruit fly Drosophila melanogaster. We found that two independent CG42260 mutants show reduced responses in the ab3A neurons. Unlike mammalian CNGs, CG42260 is not expressed in the odorant sensory neurons but broadly in the central nervous system including neuropeptide-producing cells. By using molecular genetic tools, we identified CG42260 expression in one pair of neuropeptide F (NPF) positive L1-l cells known to modulate food odor responsiveness. Knockdown of CG42260 in the NPF neurons reduced production of NPF in Ll-1 cells, which in turn, led to reduction of neuronal responses of the ab3A neurons. Our findings show the novel biological function of CG42260 in modulating olfactory responses to food odor through NPF.     

Local Enhancement of Alighting in the Melon Fly, Bactrocera cucurbitae: Effect of Olfactory, Visual, and Acoustical Stimuli

In studies conducted in Hawaii under both greenhouse and field conditions, we evaluated the propensity of melon fly females, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), to alight on either fruit mimics (agar spheres) or host fruit that were or were not occupied by conspecific resident females. We also examined the extent to which occurrence of local enhancement of alighting found in B. cucurbitae females was affected by a variety of factors such as the presence or the absence of host fruit odor (zucchini or ivy gourd), the number of conspecifics present on a host, the degree of isolation of assayed females from other females prior to testing, and the kinds of stimuli (acoustical, visual, olfactory) emanating from conspecifics present on a host mimic. In addition, we asked whether local enhancement might be operative in the food-foraging behavior of melon flies. We found that in a variety of situations, melon fly females alighted in significantly greater numbers at resources (food, fruit mimics, or host fruit) occupied by conspecific females than at unoccupied resources. Such positive influence of resident conspecific females was more pronounced in greenhouse cage assays when one or two rather than four residents were present on a host mimic (but was more pronounced when four rather than one or two residents were present on a host fruit in a field test), and was more evident when test females were grouped with conspecific females than when test females were isolated from conspecific females for 5 days before testing. Rather than acoustical or olfactory stimuli associated with resident conspecific females, the mere physical presence (visual stimulus) of a motionless dead resident melon fly female provided sufficient stimulation for test females to alight in significantly greater numbers at resources occupied by conspecific females than at unoccupied resources. We consider our findings as good evidence of local enhancement in the melon fly and discuss our results in relation to monitoring tactics for adult melon flies.     

Associative learning and memory in Drosophila: beyond olfactory conditioning

The associative learning abilities of the fruit fly, Drosophila melanogaster, have been demonstrated in both classical and operant conditioning paradigms. Efforts to identify the neural pathways and cellular mechanisms of learning have focused largely on olfactory classical conditioning. Results derived from various genetic and molecular manipulations provide considerable evidence that this form of associative learning depends critically on neural activity and cAMP signaling in brain neuropil structures called mushroom bodies. Three other behavioral learning paradigms in Drosophila serve as the main subject of this review. These are (1) visual and motor learning of flies tethered in a flight simulator, (2) a form of spatial learning that is independent of visual and olfactory cues, and (3) experience-dependent changes in male courtship behavior. The present evidence suggests that at least some of these modes of learning are independent of mushroom bodies. Applying targeted genetic manipulations to these behavioral paradigms should allow for a more comprehensive understanding of neural mechanisms responsible for diverse forms of associative learning and memory.     

Translating olfactomes into attractants: shared volatiles provide attractive bridges for polyphagy in fruit flies

Tephritid flies are serious fruit pests. Despite clear niche differences, many species show considerable overlap in fruit preferences, of which we here analysed the olfactory correlate. Using the volatiles of four unrelated fruit species, antennal responses were quantified to construct a fruit‐odour response database for four tephritid species. Although responses were distinct with a significant niche‐correlated bias, the analyses show that the probability of detection of a volatile strongly increased with its sharedness across fruits. This also held for the unrelated fruit fly Drosophila melanogaster (DoOR repository‐based analyses). We conjectured that shared volatiles signify ‘host’ to the fly ‘nose’ and induce attraction. Indeed, blends of volatiles shared by fruit and detected by all four species were very attractive for tephritid species, more than fruits. Quantitative whole antennal recordings en lieu of, or complementing bottom‐up molecular neurogenetic approaches, enables comparative olfactomics in non‐model species, and facilitate interpretation of olfaction in evolutionary, ecological, and applied contexts.