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The fruit fly, Drosophila melanogaster, is an established and powerful model system for neuroscience research with wide relevance in biology and medicine. Until recently, research on the Drosophila brain was hindered by the lack of a complete and uniform nomenclature. Recognising this problem, the Insect Brain Name Working Group produced an authoritative hierarchical nomenclature system for the adult insect brain, using Drosophila melanogaster as the reference framework, with other taxa considered to ensure greater consistency and expandability (Ito et al., 2014). Here, we extend this nomenclature system to the sub-gnathal regions of the adult Drosophila nervous system, thus providing a systematic anatomical description of the ventral nervous system (VNS). This portion of the nervous system includes the thoracic and abdominal neuromeres that were not included in the original work and contains the motor circuits that play essential roles in most fly behaviours.

To navigate through the world, animals must stabilize their path against disturbances and change direction to avoid obstacles and to search for resources [1 ; 2]. Locomotion is thus guided by sensory cues but also depends on intrinsic processes, such as motivation and physiological state. Flies, for example, turn with the direction of large-field rotatory motion, an optomotor reflex that is thought to help them fly straight [3; 4 ; 5]. Occasionally, however, they execute fast turns, called body saccades, either spontaneously or in response to patterns of visual motion such as expansion [6; 7 ; 8]. These turns can be measured in tethered flying Drosophila [ 3; 4 ; 9], which facilitates the study of underlying neural mechanisms. Whereas there is evidence for an efference copy input to visual interneurons during saccades [10], the circuits that control spontaneous and visually elicited saccades are not well known. Using two-photon calcium imaging and electrophysiological recordings in tethered flying Drosophila, we have identified a descending neuron whose activity is correlated with both spontaneous and visually elicited turns during tethered flight. The cell’s activity in open- and closed-loop experiments suggests that it does not underlie slower compensatory responses to horizontal motion but rather controls rapid changes in flight path. The activity of this neuron can explain some of the behavioral variability observed in response to visual motion and appears sufficient for eliciting turns when artificially activated. This work provides an entry point into studying the circuits underlying the control of rapid steering maneuvers in the fly brain.

Flies execute their remarkable aerial maneuvers using a set of wing steering muscles, which are activated at specific phases of the stroke cycle [1, 2, 3]. The activation phase of these muscles—which determines their biomechanical output [4, 5, 6]—arises via feedback from mechanoreceptors at the base of the wings and structures unique to flies called halteres [7, 8, 9]. Evolved from the hindwings, the tiny halteres oscillate at the same frequency as the wings, although they serve no aerodynamic function [10] and are thought to act as gyroscopes [10, 11, 12, 13, 14, 15]. Like the wings, halteres possess minute control muscles whose activity is modified by descending visual input [16], raising the possibility that flies control wing motion by adjusting the motor output of their halteres, although this hypothesis has never been directly tested. Here, using genetic techniques possible in Drosophila melanogaster, we tested the hypothesis that visual input during flight modulates haltere muscle activity and that this, in turn, alters the mechanosensory feedback that regulates the wing steering muscles. Our results suggest that rather than acting solely as a gyroscope to detect body rotation, halteres also function as an adjustable clock to set the spike timing of wing motor neurons, a specialized capability that evolved from the generic flight circuitry of their four-winged ancestors. In addition to demonstrating how the efferent control loop of a sensory structure regulates wing motion, our results provide insight into the selective scenario that gave rise to the evolution of halteres.

Insects maintain a constant bearing across a wide range of spatial scales. Monarch butterflies and locusts traverse continents [[1] and [2]], and foraging bees and ants travel hundreds of meters to return to their nests [[1], [3] and [4]], whereas many other insects fly straight for only a few centimeters before changing direction. Despite this variation in spatial scale, the brain region thought to underlie long-distance navigation is remarkably conserved [[5] and [6]], suggesting that the use of a celestial compass is a general and perhaps ancient capability of insects. Laboratory studies of Drosophila have identified a local search mode in which short, straight segments are interspersed with rapid turns [[7] and [8]]. However, this flight mode is inconsistent with measured gene flow between geographically separated populations [[9], [10] and [11]], and individual Drosophila can travel 10 km across desert terrain in a single night [[9], [12] and [13]]—a feat that would be impossible without prolonged periods of straight flight. To directly examine orientation behavior under outdoor conditions, we built a portable flight arena in which a fly viewed the natural sky through a liquid crystal device that could experimentally rotate the polarization angle. Our findings indicate that Drosophila actively orient using the sky's natural polarization pattern.

SummaryA key feature of reactive behaviors is the ability to spatially localize a salient stimulus and act accordingly. Such sensory-motor transformations must be particularly fast and well tuned in escape behaviors, in which both the speed and accuracy of the evasive response determine whether an animal successfully avoids predation [1]. We studied the escape behavior of the fruit fly, Drosophila, and found that flies can use visual information to plan a jump directly away from a looming threat. This is surprising, given the architecture of the pathway thought to mediate escape [2, 3]. Using high-speed videography, we found that approximately 200 ms before takeoff, flies begin a series of postural adjustments that determine the direction of their escape. These movements position their center of mass so that leg extension will push them away from the expanding visual stimulus. These preflight movements are not the result of a simple feed-forward motor program because their magnitude and direction depend on the flies' initial postural state. Furthermore, flies plan a takeoff direction even in instances when they choose not to jump. This sophisticated motor program is evidence for a form of rapid, visually mediated motor planning in a genetically accessible model organism.

Modern neuroscience and the interest in biomimetic control design demand increasingly sophisticated experimental techniques that can be applied in freely moving animals under realistic behavioral conditions. To explore sensorimotor flight control mechanisms in free-flying fruit flies ( Drosophila melanogaster ), we equipped a wind tunnel with a Virtual Reality (VR) display system based on standard digital hardware and a 3D path tracking system. We demonstrate the experimental power of this approach by example of a ‘one-parameter open loop’ testing paradigm. It provided (1) a straightforward measure of transient responses in presence of open loop visual stimulation; (2) high data throughput and standardized measurement conditions from process automation; and (3) simplified data analysis due to well-defined testing conditions. Being based on standard hardware and software techniques, our methods provide an affordable, easy to replicate and general solution for a broad range of behavioral applications in freely moving animals. Particular relevance for advanced behavioral research tools originates from the need to perform detailed behavioral analyses in genetically modified organisms and animal models for disease research.

Flying insects exhibit stunning behavioral repertoires that are largely mediated by the visual control of flight. For this reason, presenting a controlled visual environment to tethered insects has been and continues to be a powerful tool for studying the sensory control of complex behaviors. To create an easily controlled, scalable, and customizable visual stimulus, we have designed a modular system, based on panels composed of an 8 x 8 array of individual LEDs, that may be connected together to 'tile' an experimental environment with controllable displays. The panels have been designed to be extremely bright, with the added flexibility of individual-pixel brightness control, allowing experimentation over a broad range of behaviorally relevant conditions. Patterns to be displayed may be designed using custom software, downloaded to a controller board, and displayed on the individually addressed panels via a rapid communication interface. The panels are controlled by a microprocessor-based display controller which, for most experiments, will not require a computer in the loop, greatly reducing the experimental infrastructure. This technology allows an experimenter to build and program a visual arena with a customized geometry in a matter of hours. To demonstrate the utility of this system, we present results from experiments with tethered Drosophila melanogaster: (1) in a cylindrical arena composed of 44 panels, used to test the contrast dependence of object orientation behavior, and (2) above a 30-panel floor display, used to examine the effects of ground motion on orientation during flight.

Insects exhibit unparalleled and incredibly robust flight dynamics in the face of uncertainties. A fundamental principle contributing to this amazing behavior is rapid processing and convergence of visual sensory information to flight motor commands via spatial wide-field integration, accomplished by motion pattern sensitive interneurons in the lobula plate portion of the visual ganglia. Within a control-theoretic framework, a model for wide-field integration of retinal image flow is developed, establishing the connection between image flow kernels (retinal motion pattern sensitivities) and the feedback terms they represent. It is demonstrated that the proposed output feedback methodology is sufficient to give rise to experimentally observed navigational heuristics as the centering and forward speed regulation responses exhibited by honeybees.

Flies exhibit a repertoire of aerial acrobatics unmatched in robustness and aerodynamic sophistication. The exquisite control of this complex behavior emerges from encoding intricate patterns of optic flow, and the translation of these visual signals into the mechanical language of the motor system. Recent advances in experimental design toward more naturalistic visual and mechanosensory stimuli have served to reinforce fly flight as a key model system for understanding how feedback from multiple sensory modalities is integrated to control complex and robust motor behaviors across taxa.

The Drosophila myosin regulatory light chain (DMLC2) is homologous to MLC2s of vertebrate organisms, except for the presence of a unique 46-amino acid N-terminal extension. To study the role of the DMLC2 N-terminal extension in Drosophila flight muscle, we constructed a truncated form of the Dmlc2 gene lacking amino acids 2-46 (Dmlc2(Delta2-46)). The mutant gene was expressed in vivo, with no wild-type Dmlc2 gene expression, via P-element-mediated germline transformation. Expression of the truncated DMLC2 rescues the recessive lethality and dominant flightless phenotype of the Dmlc2 null, with no discernible effect on indirect flight muscle (IFM) sarcomere assembly. Homozygous Dmlc2(Delta2-46) flies have reduced IFM dynamic stiffness and elastic modulus at the frequency of maximum power output. The viscous modulus, a measure of the fly's ability to perform oscillatory work, was not significantly affected in Dmlc2(Delta2-46) IFM. In vivo flight performance measurements of Dmlc2(Delta2-46) flies using a visual closed-loop flight arena show deficits in maximum metabolic power (P(*)(CO(2))), mechanical power (P(*)(mech)), and flight force. However, mutant flies were capable of generating flight force levels comparable to body weight, thus enabling them to fly, albeit with diminished performance. The reduction in elastic modulus in Dmlc2(Delta2-46) skinned fibers is consistent with the N-terminal extension being a link between the thick and thin filaments that is parallel to the cross-bridges. Removal of this parallel link causes an unfavorable shift in the resonant properties of the flight system, thus leading to attenuated flight performance.

SummaryBiologists and engineers have long struggled to understand the hovering flight of insects, birds, and bats. The enormous diversity of these groups would suggest they fly using a variety of mechanisms, but a new study shows that hovering bats use the same aerodynamic mechanisms as do moths and other insects.

We examine how the structure and function of indirect flight muscle (IFM) and the entire flight system of Drosophila melanogaster are affected by phosphorylation of the myosin regulatory light chain (MLC2). This integrated study uses site-directed mutagenesis to examine the relationship between removal of the myosin light chain kinase (MLCK) phosphorylation site, in vivo function of the flight system (flight tests, wing kinematics, metabolism, power output), isolated IFM fiber mechanics, MLC2 isoform pattern, and sarcomeric ultrastructure. The MLC2 mutants exhibit graded impairment of flight ability that correlates with a reduction in both IFM and flight system power output and a reduction in the constitutive level of MLC2 phosphorylation. The MLC2 mutants have wild-type IFM sarcomere and cross-bridge structures, ruling out obvious changes in the ultrastructure as the cause of the reduced performance. We describe a viscoelastic model of cross-bridge dynamics based on sinusoidal length perturbation analysis (Nyquist plots) of skinned IFM fibers. The sinusoidal analysis suggests the high power output of Drosophila IFM required for flight results from a phosphorylation-dependent recruitment of power-generating cross-bridges rather than a change in kinetics of the power generating step. The reduction in cross-bridge number appears to affect the way mutant flies generate flight forces of sufficient magnitude to keep them airborne. In two MLC2 mutant strains that exhibit a reduced IFM power output, flies appear to compensate by lowering wingbeat frequency and by elevating wingstroke amplitude (and presumably muscle strain). This behavioral alteration is not seen in another mutant strain in which the power output and estimated number of recruited cross-bridges is similar to that of wild type.

The flight muscles of flies are separated into two physiologically, anatomically, and functionally distinct classes: power muscles and control muscles. The large indirect power muscles sustain the high level of mechanical energy required to flap the wings up and down during flight. The contractions in the asynchronous power muscles are initiated by stretch, and their slow presynaptic motor drive serves only to maintain a tonic level of cytosolic calcium. Although providing the mechanical energy for flight, the power muscles are not directly attached to the wings. Instead, their mechanical energy is transmitted to the base of the wings through the complex linkage system of the wing hinge. In contrast, the small control muscles insert directly onto the skeletal elements at the base of the wing. Through their mechanical effects on the hinge, the control muscles act collectively as a transmission system that determines how the mechanical energy produced by the power muscles is transformed into wing motion. The control muscles are activated by motor spikes in the conventional one-for-one fashion. Thus, although the control muscles can generate little mechanical power, they provide the means by which the nervous system can rapidly alter wing kinematics during sophisticated aerial maneuvers.

In primary dissociated cultures of mouse cerebellum a number of Purkinje cell-specific marker proteins and characteristic ionic currents appear at the appropriate developmental time. During the first week after plating, Purkinje cell dendrites elongate, but as electrical activity emerges the dendrites stop growing and branch. If endogenous electrical activity is inhibited by chronic tetrodotoxin or high magnesium treatment, dendrites continue to elongate, as if they were still immature. At the time that branching begins, intracellular calcium levels become sensitive to tetrodotoxin, suggesting that this cation may be involved in dendrite growth. Even apparently mature Purkinje cells alter their dendritic growth in response to changes in activity, suggesting long-term plasticity.

The neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) is generally believed to selectively ablate serotonergic neurons. Serotonin-containing Retzius cells (RZ) in the leech appear ablated by 5,7-DHT treatment. However, these brown, mis-shapen, non-fluorescent neurons retain their synaptic inputs, action potentials, electronic outputs, a membrane receptor, and axonal projections.