Asynchronous haltere input drives specific wing and head movements in Drosophila.

Halteres are multifunctional mechanosensory organs unique to the true flies (Diptera). A set of reduced hindwings, the halteres beat at the same frequency as the lift-generating forewings and sense inertial forces via mechanosensory campaniform sensilla. Though haltere ablation makes stable flight impossible, the specific role of wing-synchronous input has not been established. Using small iron filings attached to the halteres of tethered flies and an alternating electromagnetic field, we experimentally decoupled the wings and halteres of flying Drosophila and observed the resulting changes in wingbeat amplitude and head orientation. We find that asynchronous haltere input results in fast amplitude changes in the wing (hitches), but does not appreciably move the head. In multi-modal experiments, we find that wing and gaze optomotor responses are disrupted differently by asynchronous input. These effects of wing-asynchronous haltere input suggest that specific sensory information is necessary for maintaining wing amplitude stability and adaptive gaze control.

Fantastic beasts and how to study them: rethinking experimental animal behavior.

Humans have been trying to understand animal behavior at least since recorded history. Recent rapid development of new technologies has allowed us to make significant progress in understanding the physiological and molecular mechanisms underlying behavior, a key goal of neuroethology. However, there is a tradeoff when studying animal behavior and its underlying biological mechanisms: common behavior protocols in the laboratory are designed to be replicable and controlled, but they often fail to encompass the variability and breadth of natural behavior. This Commentary proposes a framework of 10 key questions that aim to guide researchers in incorporating a rich natural context into their experimental design or in choosing a new animal study system. The 10 questions cover overarching experimental considerations that can provide a template for interspecies comparisons, enable us to develop studies in new model organisms and unlock new experiments in our quest to understand behavior.

Haltere and visual inputs sum linearly to predict wing (but not gaze) motor output in tethered flying Drosophila.

In the true flies (Diptera), the hind wings have evolved into specialized mechanosensory organs known as halteres, which are sensitive to gyroscopic and other inertial forces. Together with the fly's visual system, the halteres direct head and wing movements through a suite of equilibrium reflexes that are crucial to the fly's ability to maintain stable flight. As in other animals (including humans), this presents challenges to the nervous system as equilibrium reflexes driven by the inertial sensory system must be integrated with those driven by the visual system in order to control an overlapping pool of motor outputs shared between the two of them. Here, we introduce an experimental paradigm for reproducibly altering haltere stroke kinematics and use it to quantify multisensory integration of wing and gaze equilibrium reflexes. We show that multisensory wing-steering responses reflect a linear superposition of haltere-driven and visually driven responses, but that multisensory gaze responses are not well predicted by this framework. These models, based on populations, extend also to the responses of individual flies.

Takeoff diversity in Diptera.

The order Diptera (true flies) are named for their two wings because their hindwings have evolved into specialized mechanosensory organs called halteres. Flies use halteres to detect body rotations and maintain stability during flight and other behaviours. The most recently diverged dipteran monophyletic subsection, the Calyptratae, is highly successful, accounting for approximately 12% of dipteran diversity, and includes common families like house flies. These flies move their halteres independently from their wings and oscillate their halteres during walking. Here, we demonstrate that this subsection of flies uses their halteres to stabilize their bodies during takeoff, whereas non-Calyptratae flies do not. We find that flies of the Calyptratae are able to take off more rapidly than non-Calyptratae flies without sacrificing stability. Haltere removal decreased both velocity and stability in the takeoffs of Calyptratae, but not other flies. The loss of takeoff velocity following haltere removal in Calyptratae (but not other flies) is a direct result of a decrease in leg extension speed. A closely related non-Calyptratae species (D. melanogaster) also has a rapid takeoff, but takeoff duration and stability are unaffected by haltere removal. Haltere use thus allows for greater speed and stability during fast escapes, but only in the Calyptratae clade.

Representation of Haltere Oscillations and Integration with Visual Inputs in the Fly Central Complex.

The reduced hindwings of flies, known as halteres, are specialized mechanosensory organs that detect body rotations during flight. Primary afferents of the haltere encode its oscillation frequency linearly over a wide bandwidth and with precise phase-dependent spiking. However, it is not currently known whether information from haltere primary afferent neurons is sent to higher brain centers where sensory information about body position could be used in decision making, or whether precise spike timing is useful beyond the peripheral circuits that drive wing movements. We show that in cells in the central brain, the timing and rates of neural spiking can be modulated by sensory input from experimental haltere movements (driven by a servomotor). Using multichannel extracellular recording in restrained flesh flies (Sarcophaga bullata of both sexes), we examined responses of central complex cells to a range of haltere oscillation frequencies alone, and in combination with visual motion speeds and directions. Haltere-responsive units fell into multiple response classes, including those responding to any haltere motion and others with firing rates linearly related to the haltere frequency. Cells with multisensory responses showed higher firing rates than the sum of the unisensory responses at higher haltere frequencies. They also maintained visual properties, such as directional selectivity, while increasing response gain nonlinearly with haltere frequency. Although haltere inputs have been described extensively in the context of rapid locomotion control, we find haltere sensory information in a brain region known to be involved in slower, higher-order behaviors, such as navigation.SIGNIFICANCE STATEMENT Many animals use vision for navigation; however, these cues must be interpreted in the context of the body's position. In mammalian brains, hippocampal cells combine visual and vestibular information to encode head direction. A region of the arthropod brain, known as the central complex (CX), similarly encodes heading information, but it is unknown whether proprioceptive information is integrated here as well. We show that CX neurons respond to input from halteres, specialized proprioceptors in flies that detect body rotations. These neurons also respond to visual input, providing one of the few examples of multiple sensory modalities represented in individual CX cells. Haltere stimulation modifies neural responses to visual signals, providing a mechanism for integrating vision with proprioception.

Single mechanosensory neurons encode lateral displacements using precise spike timing and thresholds.

During locomotion, animals rely on multiple sensory modalities to maintain stability. External cues may guide behaviour, but they must be interpreted in the context of the animal's own body movements. Mechanosensory cues that can resolve dynamic internal and environmental conditions, like those from vertebrate vestibular systems or other proprioceptors, are essential for guided movement. How do afferent proprioceptor neurons transform movement into a neural code? In flies, modified hindwings known as halteres detect forces produced by body rotations and are essential for flight. However, the mechanisms by which haltere neurons transform forces resulting from three-dimensional body rotations into patterns of neural spikes are unknown. We use intracellular electrodes to record from haltere primary afferent neurons during a range of haltere motions. We find that spike timing activity of individual neurons changes with displacement and propose a mechanism by which single neurons can encode three-dimensional haltere movements during flight.

Haltere removal alters responses to gravity in standing flies.

Animals detect the force of gravity with multiple sensory organs, from subcutaneous receptors at body joints to specialized sensors like the vertebrate inner ear. The halteres of flies, specialized mechanoreceptive organs derived from hindwings, are known to detect body rotations during flight, and some groups of flies also oscillate their halteres while walking. The dynamics of halteres are such that they could act as gravity detectors for flies standing on substrates, but their utility during non-flight behaviors is not known. We observed the behaviors of intact and haltere-ablated flies during walking and during perturbations in which the acceleration due to gravity suddenly changed. We found that intact halteres are necessary for flies to maintain normal walking speeds on vertical surfaces and to respond to sudden changes in gravity. Our results suggest that halteres can serve multiple sensory purposes during different behaviors, expanding their role beyond their canonical use in flight.

Cross-modal influence of mechanosensory input on gaze responses to visual motion in Drosophila.

Animals typically combine inertial and visual information to stabilize their gaze against confounding self-generated visual motion, and to maintain a level gaze when the body is perturbed by external forces. In vertebrates, an inner ear vestibular system provides information about body rotations and accelerations, but gaze stabilization is less understood in insects, which lack a vestibular organ. In flies, the halteres, reduced hindwings imbued with hundreds of mechanosensory cells, sense inertial forces and provide input to neck motoneurons that control gaze. These neck motoneurons also receive input from the visual system. Head movement responses to visual motion and physical rotations of the body have been measured independently, but how inertial information might influence gaze responses to visual motion has not been fully explored. We measured the head movement responses to visual motion in intact and haltere-ablated tethered flies to explore the role of the halteres in modulating visually guided head movements in the absence of rotation. We note that visually guided head movements occur only during flight. Although halteres are not necessary for head movements, the amplitude of the response is smaller in haltereless flies at higher speeds of visual motion. This modulation occurred in the absence of rotational body movements, demonstrating that the inertial forces associated with straight tethered flight are important for gaze-control behavior. The cross-modal influence of halteres on the fly's responses to fast visual motion indicates that the haltere's role in gaze stabilization extends beyond its canonical function as a sensor of angular rotations of the thorax.

Haltere mechanosensory influence on tethered flight behavior in Drosophila.

In flies, mechanosensory information from modified hindwings known as halteres is combined with visual information for wing-steering behavior. Haltere input is necessary for free flight, making it difficult to study the effects of haltere ablation under natural flight conditions. We thus used tethered Drosophila melanogaster flies to examine the relationship between halteres and the visual system, using wide-field motion or moving figures as visual stimuli. Haltere input was altered by surgically decreasing its mass, or by removing it entirely. Haltere removal does not affect the flies' ability to flap or steer their wings, but it does increase the temporal frequency at which they modify their wingbeat amplitude. Reducing the haltere mass decreases the optomotor reflex response to wide-field motion, and removing the haltere entirely does not further decrease the response. Decreasing the mass does not attenuate the response to figure motion, but removing the entire haltere does attenuate the response. When flies are allowed to control a visual stimulus in closed-loop conditions, haltereless flies fixate figures with the same acuity as intact flies, but cannot stabilize a wide-field stimulus as accurately as intact flies can. These manipulations suggest that the haltere mass is influential in wide-field stabilization, but less so in figure tracking. In both figure and wide-field experiments, we observe responses to visual motion with and without halteres, indicating that during tethered flight, intact halteres are not strictly necessary for visually guided wing-steering responses. However, the haltere feedback loop may operate in a context-dependent way to modulate responses to visual motion.

Kinematic diversity suggests expanded roles for fly halteres.

The halteres of flies are mechanosensory organs that provide information about body rotations during flight. We measured haltere movements in a range of fly taxa during free walking and tethered flight. We find a diversity of wing-haltere phase relationships in flight, with higher variability in more ancient families and less in more derived families. Diverse haltere movements were observed during free walking and were correlated with phylogeny. We predicted that haltere removal might decrease behavioural performance in those flies that move them during walking and provide evidence that this is the case. Our comparative approach reveals previously unknown diversity in haltere movements and opens the possibility of multiple functional roles for halteres in different fly behaviours.

Figure-ground discrimination behavior in Drosophila. II. Visual influences on head movement behavior.

Visual identification of small moving targets is a challenge for all moving animals. Their own motion generates displacement of the visual surroundings, inducing wide-field optic flow across the retina. Wide-field optic flow is used to sense perturbations in the flight course. Both ego-motion and corrective optomotor responses confound any attempt to track a salient target moving independently of the visual surroundings. What are the strategies that flying animals use to discriminate small-field figure motion from superimposed wide-field background motion? We examined how fruit flies adjust their gaze in response to a compound visual stimulus comprising a small moving figure against an independently moving wide-field ground, which they do by re-orienting their head or their flight trajectory. We found that fixing the head in place impairs object fixation in the presence of ground motion, and that head movements are necessary for stabilizing wing steering responses to wide-field ground motion when a figure is present. When a figure is moving relative to a moving ground, wing steering responses follow components of both the figure and ground trajectories, but head movements follow only the ground motion. To our knowledge, this is the first demonstration that wing responses can be uncoupled from head responses and that the two follow distinct trajectories in the case of simultaneous figure and ground motion. These results suggest that whereas figure tracking by wing kinematics is independent of head movements, head movements are important for stabilizing ground motion during active figure tracking.

Figure-ground discrimination behavior in Drosophila. I. Spatial organization of wing-steering responses.

The behavioral algorithms and neural subsystems for visual figure-ground discrimination are not sufficiently described in any model system. The fly visual system shares structural and functional similarity with that of vertebrates and, like vertebrates, flies robustly track visual figures in the face of ground motion. This computation is crucial for animals that pursue salient objects under the high performance requirements imposed by flight behavior. Flies smoothly track small objects and use wide-field optic flow to maintain flight-stabilizing optomotor reflexes. The spatial and temporal properties of visual figure tracking and wide-field stabilization have been characterized in flies, but how the two systems interact spatially to allow flies to actively track figures against a moving ground has not. We took a systems identification approach in flying Drosophila and measured wing-steering responses to velocity impulses of figure and ground motion independently. We constructed a spatiotemporal action field (STAF) - the behavioral analog of a spatiotemporal receptive field - revealing how the behavioral impulse responses to figure tracking and concurrent ground stabilization vary for figure motion centered at each location across the visual azimuth. The figure tracking and ground stabilization STAFs show distinct spatial tuning and temporal dynamics, confirming the independence of the two systems. When the figure tracking system is activated by a narrow vertical bar moving within the frontal field of view, ground motion is essentially ignored despite comprising over 90% of the total visual input.

Flies tune the activity of their multifunctional gyroscope.

Members of the order Diptera, the true flies, are among the most maneuverable flying animals. These aerial capabilities are partially attributed to flies' possession of halteres, tiny club-shaped structures that evolved from the hindwings and play a crucial role in flight control. Halteres are renowned for acting as biological gyroscopes that rapidly detect rotational perturbations and help flies maintain a stable flight posture. Additionally, halteres provide rhythmic input to the wing steering system that can be indirectly modulated by the visual system. The multifunctional capacity of the haltere is thought to depend on arrays of embedded mechanosensors called campaniform sensilla that are arranged in distinct groups on the haltere's dorsal and ventral surfaces. Although longstanding hypotheses suggest that each array provides different information relevant to the flight control circuitry, we know little about how the haltere campaniforms are functionally organized. Here, we use in vivo calcium imaging during tethered flight to obtain population-level recordings of the haltere sensory afferents in specific fields of sensilla. We find that haltere feedback from both dorsal fields is continuously active, modulated under closed-loop flight conditions, and recruited during saccades to help flies actively maneuver. We also find that the haltere's multifaceted role may arise from the steering muscles of the haltere itself, regulating haltere stroke amplitude to modulate campaniform activity. Taken together, our results underscore the crucial role of efferent control in regulating sensor activity and provide insight into how the sensory and motor systems of flies coevolved.

Encoding properties of haltere neurons enable motion feature detection in a biological gyroscope.

The halteres of dipteran insects are essential sensory organs for flight control. They are believed to detect Coriolis and other inertial forces associated with body rotation during flight. Flies use this information for rapid flight control. We show that the primary afferent neurons of the haltere's mechanoreceptors respond selectively with high temporal precision to multiple stimulus features. Although we are able to identify many stimulus features contributing to the response using principal component analysis, predictive models using only two features, common across the cell population, capture most of the cells' encoding activity. However, different sensitivity to these two features permits each cell to respond to sinusoidal stimuli with a different preferred phase. This feature similarity, combined with diverse phase encoding, allows the haltere to transmit information at a high rate about numerous inertial forces, including Coriolis forces.

Insect neuroethology: Flight constructs a compass for monarch migration.

A new study shows that a brain map in the monarch butterfly can be re-drawn during flight. Migrating butterflies integrate efferent and visual signals to create an adaptable compass in their central brain.

Inertial Sensing and Encoding of Self-Motion: Structural and Functional Similarities across Metazoan Taxa.

To properly orient and navigate, moving animals must obtain information about the position and motion of their bodies. Animals detect inertial signals resulting from body accelerations and rotations using a variety of sensory systems. In this review, we briefly summarize current knowledge on inertial sensing across widely disparate animal taxa with an emphasis on neuronal coding and sensory transduction. We outline systems built around mechanosensory hair cells, including the chordate vestibular complex and the statocysts seen in many marine invertebrates. We next compare these to schemes employed by flying insects for managing inherently unstable aspects of flapping flight, built around comparable mechanosensory cells but taking unique advantage of the physics of rotating systems to facilitate motion encoding. Finally, we highlight fundamental similarities across taxa with respect to the partnering of inertial senses with visual senses and conclude with a discussion of the functional utility of maintaining a multiplicity of encoding schemes for self-motion information.

Sensory Feedback and Animal Locomotion: Perspectives from Biology and Biorobotics: An Introduction to the Symposium.

The successful completion of many behaviors relies on sensory feedback. This symposium brought together researchers using novel techniques to study how different stimuli are encoded, how and where multimodal feedback is integrated, and how feedback modulates motor output in diverse modes of locomotion (aerial, aquatic, and terrestrial) in a diverse range of taxa (insects, fish, tetrapods), and in robots. Similar to biological organisms, robots can be equipped with integrated sensors and can rely on sensory feedback to adjust the output signal of a controller. Engineers often look to biology for inspiration on how animals have evolved solutions to problems similar to those experienced in robotic movement. Similarly, biologists too must proactively engage with engineers to apply computer and robotic models to test hypotheses and answer questions on the capacity and roles of sensory feedback in generating effective movement. Through a diverse group of researchers, including both biologists and engineers, the symposium attempted to catalyze new interdisciplinary collaborations and identify future research directions for the development of bioinspired sensory control systems, as well as the use of robots to test hypotheses in neuromechanics.

Haltere morphology and campaniform sensilla arrangement across Diptera.

One of the primary specializations of true flies (order Diptera) is the modification of the hind wings into club-shaped halteres. Halteres are complex mechanosensory structures that provide sensory feedback essential for stable flight control via an array of campaniform sensilla at the haltere base. The morphology of these sensilla has previously been described in a small number of dipteran species, but little is known about how they vary across fly taxa. Using a synoptic set of specimens representing 42 families from all of the major infraorders of Diptera, we used scanning electron microscopy to map the gross and fine structures of halteres, including sensillum shape and arrangement. We found that several features of haltere morphology correspond with dipteran phylogeny: Schizophora generally have smaller halteres with stereotyped and highly organized sensilla compared to nematoceran flies. We also found a previously undocumented high variation of haltere sensillum shape in nematoceran dipterans, as well as the absence of a dorsal sensillum field in multiple families. Overall, variation in haltere sensillar morphology across the dipteran phylogeny provides insight into the evolution of a highly specialized proprioceptive organ and a basis for future studies on haltere sensory function.

Dipteran Halteres: Perspectives on Function and Integration for a Unique Sensory Organ.

The halteres of dipteran insects (true flies) are essential mechanosensory organs for flight. These are modified hindwings with several arrays of sensory cells at their base, and they are one of the characteristic features of flies. Mechanosensory information from the halteres is sent with low latency to wing-steering and head movement motoneurons, allowing direct control of body position and gaze. Analyses of the structure and dynamics of halteres indicate that they experience very small aerodynamic forces but significant inertial forces, including Coriolis forces associated with body rotations. The sensory cells at the base of the haltere detect these forces and allow the fly to correct for perturbations during flight, but new evidence suggests that this may not be their only role. This review will examine our current understanding of how these organs move, encode forces, and transmit information about these forces to the nervous system to guide behavior.

Animal behavior: fly flight moves forward.

A new study has resolved the paradox of how flies maintain reflexive aversion to your approaching swatter, whilst tolerating similar visual signals during normal forward flight.