Activation phase ensures kinematic efficacy in flight-steering muscles of Drosophila melanogaster

During tethered flight in Drosophila melanogaster, spike activity of the second basalar flight-control muscle (M.b2) is correlated with an increase in both the ipsilateral wing beat amplitude and the ipsilateral flight force. The frequency of muscle spikes within a burst is about 100 Hz, or 1 spike for every two wing beat cycles. When M.b2 is active, its spikes tend to occur within a comparatively narrow phase band of the wing beat cycle. To understand the functional role of this phase-lock of firing in the control of flight forces, we stimulated M.b2 in selected phases of the wing beat cycle and recorded the effect on the ipsilateral wing beat amplitude. Varying the phase timing of the stimulus had a significant effect on the wing beat amplitude. A maximum increase of wing beat amplitude was obtained by stimulating M.b2 at the beginning of the upstroke or about 1 ms prior to the narrow phase band in which the muscle spikes typically occur during flight. Assuming a delay of 1 ms between the stimulation of the motor nerve and muscle activation, these results indicate that M.b2 is activated at an instant of the stroke cycle that produces the greatest effect on wing beat amplitude.     

Myosin light chain-2 mutation affects flight, wing beat frequency, and indirect flight muscle contraction kinetics in Drosophila.

We have used a combination of classical genetic, molecular genetic, histological, biochemical, and biophysical techniques to identify and characterize a null mutation of the myosin light chain-2 (MLC-2) locus of Drosophila melanogaster. Mlc2E38 is a null mutation of the MLC-2 gene resulting from a nonsense mutation at the tenth codon position. Mlc2E38 confers dominant flightless behavior that is associated with reduced wing beat frequency. Mlc2E38 heterozygotes exhibit a 50% reduction of MLC-2 mRNA concentration in adult thoracic musculature, which results in a commensurate reduction of MLC-2 protein in the indirect flight muscles. Indirect flight muscle myofibrils from Mlc2E38 heterozygotes are aberrant, exhibiting myofilaments in disarray at the periphery. Calcium-activated Triton X-100-treated single fiber segments exhibit slower contraction kinetics than wild type. Introduction of a transformed copy of the wild type MLC-2 gene rescues the dominant flightless behavior of Mlc2E38 heterozygotes. Wing beat frequency and single fiber contraction kinetics of a representative rescued line are not significantly different from those of wild type. Together, these results indicate that wild type MLC-2 stoichiometry is required for normal indirect flight muscle assembly and function. Furthermore, these results suggest that the reduced wing beat frequency and possibly the flightless behavior conferred by Mlc2E38 is due in part to slower contraction kinetics of sarcomeric regions devoid or partly deficient in MLC-2.     

Alternative N-terminal regions of Drosophila myosin heavy chain tune muscle kinetics for optimal power output

We assessed the influence of alternative versions of a region near the N-terminus of Drosophila myosin heavy chain on muscle mechanical properties. Previously, we exchanged N-terminal regions (encoded by alternative exon 3s) between an embryonic (EMB) isoform and the indirect flight muscle isoform (IFI) of myosin, and demonstrated that it influences solution ATPase rates and in vitro actin sliding velocity. Because each myosin is expressed in Drosophila indirect flight muscle, in the absence of other myosin isoforms, this allows for muscle mechanical and whole organism locomotion assays. We found that exchanging the flight muscle specific exon 3 region into the embryonic isoform (EMB-3b) increased maximum power generation (P(max)) and optimal frequency of power generation (f(max)) threefold and twofold compared to fibers expressing EMB, whereas exchanging the embryonic exon 3 region into the flight muscle isoform (IFI-3a) decreased P(max) and f(max) to approximately 80% of IFI fiber values. Drosophila expressing IFI-3a exhibited a reduced wing beat frequency compared to flies expressing IFI, which optimized power generation from their kinetically slowed flight muscle. However, the slower wing beat frequency resulted in a substantial loss of aerodynamic power as manifest in decreased flight performance of IFI-3a compared to IFI. Thus the N-terminal region is important in tuning myosin kinetics to match muscle speed for optimal locomotory performance.     

The myosin filament superlattice in the flight muscles of flies: A-band lattice optimisation for stretch-activation?

Low-angle X-ray diffraction patterns from relaxed fruitfly (Drosophila) flight muscle recorded on the BioCat beamline at the Argonne Advanced Photon Source (APS) show many features similar to such patterns from the "classic" insect flight muscle in Lethocerus, the giant water bug, but there is a characteristically different pattern of sampling of the myosin filament layer-lines, which indicates the presence of a superlattice of myosin filaments in the Drosophila A-band. We show from analysis of the structure factor for this lattice that the sampling pattern is exactly as expected if adjacent four-stranded myosin filaments, of repeat 116 nm, are axially shifted in the hexagonal A-band lattice by one-third of the 14.5 nm axial spacing between crowns of myosin heads. In addition, electron micrographs of Drosophila and other flies (e.g. the house fly (Musca) and the flesh fly (Sarcophaga)) combined with image processing confirm that the same A-band superlattice occurs in all of these flies; it may be a general property of the Diptera. The different A-band organisation in flies compared with Lethocerus, which operates at a much lower wing beat frequency (approximately 30 Hz) and requires a warm-up period, may be a way of optimising the myosin and actin filament geometry needed both for stretch activation at the higher wing beat frequencies (50 Hz to 1000 Hz) of flies and their need for a rapid escape response.     

Wing‐beat characteristics of birds recorded with tracking radar and cine camera

This study presents wing‐beat frequency data measured mainly by radar, complemented by video and cinematic recordings, for 153 western Palaearctic and two African species. Data on a further 45 Palaearctic species from other sources are provided in an electronic appendix. For 41 species with passerine‐type flight, the duration of flapping and pausing phases is given. The graphical presentations of frequency ranges and wing‐beat patterns show within‐species variation and allow easy comparison between species, taxonomic groups and types of flight. Wing‐beat frequency is described by Pennycuick (J. Exp. Biol. 2001; 204: 3283–3294) as a function of body‐mass, wing‐span, wing‐area, gravity and air density; for birds with passerine‐type flight the power‐fraction has also to be considered. We tested Pennycuick’s general allometric model and estimated the coefficients based on our data. The general model explained a high proportion of variation in wing‐beat frequency and the coefficients differed only slightly from Pennycuick’s original values. Modelling continuous‐flapping flyers alone resulted in coefficients not different from those predicted (within 95% intervals). Doing so for passerine‐type birds resulted in a model with non‐significant contributions of body‐mass and wing‐span to the model. This was mainly due to the very high correlation between body‐mass, wing‐span and wing‐area, revealing similar relative scaling properties within this flight type. However, wing‐beat frequency increased less than expected with respect to power‐fraction, indicating that the drop in flight level during the non‐flapping phases, compensated by the factor (g/q)^0.5 in Pennycuick’s model, is smaller than presumed. This may be due to lift produced by the body during the bounding phase or by only partial folding of the wings.     

Morphological and kinematic basis of the hummingbird flight stroke: scaling of flight muscle transmission ratio

Hummingbirds (Trochilidae) are widely known for their insect-like flight strokes characterized by high wing beat frequency, small muscle strains and a highly supinated wing orientation during upstroke that allows for lift production in both halves of the stroke cycle. Here, we show that hummingbirds achieve these functional traits within the limits imposed by a vertebrate endoskeleton and muscle physiology by accentuating a wing inversion mechanism found in other birds and using long-axis rotational movement of the humerus. In hummingbirds, long-axis rotation of the humerus creates additional wing translational movement, supplementing that produced by the humeral elevation and depression movements of a typical avian flight stroke. This adaptation increases the wing-to-muscle-transmission ratio, and is emblematic of a widespread scaling trend among flying animals whereby wing-to-muscle-transmission ratio varies inversely with mass, allowing animals of vastly different sizes to accommodate aerodynamic, biomechanical and physiological constraints on muscle-powered flapping flight.     

On the sex-specific mechanisms of butterfly flight: flight performance relative to flight morphology, wing kinematics, and sex in Pararge aegeria

Many evolutionary ecological studies have documented sexual dimorphism in morphology or behaviour. However, to what extent a sex-specific morphology is used differently to realize a certain level of behavioural performance is only rarely tested. We experimentally quantified flight performance and wing kinematics (wing beat frequency and wing stroke amplitude) and flight morphology (thorax mass, body mass, forewing aspect ratio, and distance to centre of forewing area) in the butterfly Pararge aegeria (L.) using a tethered tarsal reflex induced flight set-up under laboratory conditions. On average, females showed higher flight performance than males, but frequency and amplitude did not differ. In both sexes, higher flight performance was partly determined by wing beat frequency but not by wing stroke amplitude. Dry body mass, thorax mass, and distance to centre of forewing area were negatively related to wing beat frequency. The relationship between aspect ratio and wing stroke amplitude was sex-specific: females with narrower wings produced higher amplitude whereas males show the opposite pattern. The results are discussed in relation to sexual differences in flight behaviour.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 675–687.     

Evolution of the wave: aerodynamic and aposematic functions of butterfly wing motion

Many unpalatable butterfly species use coloration to signal their distastefulness to birds, but motion cues may also be crucial to ward off predatory attacks. In previous research, captive passion-vine butterflies Heliconius mimetic in colour pattern were also mimetic in motion. Here, I investigate whether wing motion changes with the flight demands of different behaviours. If birds select for wing motion as a warning signal, aposematic butterflies should maintain wing motion independently of behavioural context. Members of one mimicry group (Heliconius cydno and Heliconius sapho) beat their wings more slowly and their wing strokes were more asymmetric than their sister-species (Heliconius melpomene and Heliconius erato, respectively), which were members of another mimicry group having a quick and steady wing motion. Within mimicry groups, wing beat frequency declined as its role in generating lift also declined in different behavioural contexts. In contrast, asymmetry of the stroke was not associated with wing beat frequency or behavioural context-strong indication that birds process and store the Fourier motion energy of butterfly wings. Although direct evidence that birds respond to subtle differences in butterfly wing motion is lacking, birds appear to generalize a motion pattern as much as they encounter members of a mimicry group in different behavioural contexts.     

Exploring the relationship between flapping behaviour and accelerometer signal during ascending flight,and a new approach to calibration

We understand little about the energetic costs of flight in free-ranging birds, in part because current techniques for estimating flight energetics in the wild are limited. Accelerometry is known to estimate energy expenditure through body movement in terrestrial animals, once calibrated using a treadmill with chamber respirometry. The flight equivalent, a wind tunnel with mask respirometry, is particularly difficult to instigate, and has not been applied to calibrate accelerometry. We take the first steps in exploring a novel method for calibrating accelerometers with flight energy expenditure. We collected accelerometry data for Harris's Hawks Parabuteo unicinctus flying to varying heights up to 4.1 m over a small horizontal distance; the mechanical energy expended to gain height can be estimated from physical first principles. The relationship between accelerometry and mechanical energy expenditure was strong, and while a simple wing flapping model confirmed that accelerometry is sensitive to both changes in wing beat amplitude and frequency, the relationship was explained predominately by changes in wing beat frequency, and less so by changes in amplitude. Our study provides initial, positive evidence that accelerometry can be calibrated with body power using climbing flights, potentially providing a basis for estimating flapping flight metabolic rate at least in situations of altitude gain.     

Avian wing proportions and flight styles: first step towards predicting the flight modes of mesozoic birds

We investigated the relationship between wing element proportions and flight mode in a dataset of living avian species to provide a framework for making basic estimates of the range of flight styles evolved by Mesozoic birds. Our results show that feather length (f(prim)) and total arm length (ta) (sum of the humerus, ulna and manus length) ratios differ significantly between four flight style groups defined and widely used for living birds and as a result are predictive for fossils. This was confirmed using multivariate ordination analyses, with four wing elements (humerus, ulna/radius, manus, primary feathers), that discriminate the four broad flight styles within living birds. Among the variables tested, manus length is closely correlated with wing size, yet is the poorest predictor for flight style, suggesting that the shape of the bones in the hand wing is most important in determining flight style. Wing bone thickness (shape) must vary with wing beat strength, with weaker forces requiring less bone. Finally, we show that by incorporating data from Mesozoic birds, multivariate ordination analyses can be used to predict the flight styles of fossils.     

Courtship song analysis of Drosophila muscle mutants

As part of the mating ritual, males of Drosophila species produce species-specific courtship songs through wing vibrations generated by the thoracic musculature. While previous studies have shown that indirect flight muscles (IFM) are neurally activated during courtship song production, the precise role of these muscles in song production has not been investigated. Fortunately, IFM mutants abound in Drosophila melanogaster and studies spanning several decades have shed light on the role of muscle proteins in IFM-powered flight. Analysis of courtship songs in these mutants offers the opportunity to uncover the role of the IFM in a behavior distinct than flight and subject to different evolutionary selection regimes. Here, we describe protocols for the recording and analysis of courtship behavior and mating song of D. melanogaster muscle transgenic and mutant strains. To record faint acoustic signal of courtship songs, an insulated mating compartment was used inside a recording device (INSECTAVOX) equipped with a modified electret microphone, a low-noise power supply, and noise filters. Songs recorded in the INSECTAVOX are digitized using Goldwave, whose several features enable extraction of critical song parameters, including carrier frequencies for pulse song and sine song. We demonstrate the utility of this approach by showing that deletion of the N-terminal region of the myosin regulatory light chain, a mutation known to decrease wing beat frequency and flight power, affects courtship song parameters.     

The myosin converter domain modulates muscle performance

Myosin is the molecular motor that powers muscle contraction as a result of conformational changes during its mechanochemical cycle. We demonstrate that the converter, a compact structural domain that differs in sequence between Drosophila melanogaster myosin isoforms, dramatically influences the kinetic properties of myosin and muscle fibres. Transgenic replacement of the converter in the fast indirect flight muscle with the converter from an embryonic muscle slowed muscle kinetics, forcing a compensatory reduction in wing beat frequency to sustain flight. Conversely, replacing the embryonic converter with the flight muscle converter sped up muscle kinetics and increased maximum power twofold, compared to flight muscles expressing the embryonic myosin isoform. The substitutions also dramatically influenced in vitro actin sliding velocity, suggesting that the converter modulates a rate-limiting step preceding cross-bridge detachment. Our integrative analysis demonstrates that isoform-specific differences in the myosin converter allow different muscle types to meet their specific locomotion demands.     

The kinematics of Heteroptera in free flight

The kinematics of six species of Heteroptera in free flight are analysed and compared.

• (1) 

  Using nested analysis of variance techniques, statistically significant variation was detected between species for several of the flight parameters measured: mean angular velocity; pronation/supination ratio; upstroke/downstroke ratio; and wing beat frequency. In each case this is discussed in terms of variation in flight behaviour.
• (2) 

  Beneficial aerodynamic forces are generated during the upstroke and the downstroke, in both fast forward and rising flight.
• (3). 

  When the insects change from level, forward flight to near vertical, rising flight, the following parameters are altered in most of the sequences analysed:

• (a). 

  the stroke plane angle becomes steeply, negatively inclined, associated with an increase in body angle;
• (b). 

  the stroke amplitude is reduced;
• (c). 

  wing beat frequency is lowered, associated with a drop in mean angular velocity;
• (d). 

  the speed of stroke reversal (rotational velocity) is increased. This may be associated with increased wing torsion and tip flexion which in turn could improve any beneficial unsteady aerodynamic effects generated at stroke reversal.

The reasons for this change in flight performance and the deviations from that seen in other insects are discussed.
It is shown that Heteroptera may make use of wing drag in flight, particularly during rising flight.     

Synchronization of wing beat cycle of the desert locust, Schistocerca gregaria, by periodic light flashes

Studies on the generation of rhythmic motor patterns have shown that peripheral sensory input may contribute substantially to the rhythm generating network. A prominent example is the wing beat frequency of desert locusts, which can be entrained to rhythmic mechanosensory input, but also to the frequency of periodic light flashes. To further analyze the entrainment by light, tethered flying locusts were presented with periodic light flashes, while the position of the forewing was filmed. We show that entrainment of wing beat occurs both in the UV and green range of light. Animals maintained a characteristic phase relationship to the light stimulus with the most elevated wing position occurring at the end of the dark phase. Speed and time course of entrainment varied greatly and ranged from the duration of a single wing beat cycle to several seconds. To identify the visual system mediating entrainment, synchronization to UV light was tested after cutting the optic stalks to the optic lobes/compound eyes or the ocellar nerves. The results show that light entrainment of the locust flight pattern is largely and perhaps exclusively mediated via the fast ocellar pathway and may have a role to stabilize flight with respect to the horizon.     

Analysis of cell vibration for assessing axonemal motility in Chlamydomonas

Flagellar mutants of Chlamydomonas have greatly contributed to our understanding of the function of axonemes and axonemal dyneins. An important step in studying mutants is to correlate the molecular and structural defects in the axoneme with motility. This is not always easy, however, partly because it is often necessary to quantify axonemal motility by measuring the cell's swimming velocity, the flagellar beat frequency, or flagellar waveform in a number of cells or axonemes. To skip this time-consuming step, a quick method for measuring the average flagellar beat frequency in a population of cells is developed based on fast Fourier transform (FFT) analysis of the vibration of cell bodies. This method yields the average beat frequency within 10-60 s and has been used as a powerful tool for identifying mutants lacking various dynein species. It is also particularly useful for studies analyzing detergent-extracted cell models under various reactivation conditions.     

The implications of function on the origin and homologies of the dipterous wing

Abstract The origin of Diptera, and the homologies of the dipteran wing, are re-examined in the light of recent studies on the flight biomechanics and functional wing morphology of Diptera and of Panorpa. Significant Diptera apomorphies are identified, relevant fossils discussed, and a hypothetical wing ground-plan figured.
The arculus, the modified clavus and the anteroposterior asymmetry of the fly wing seem to be adaptations to a mode of flight in which instantaneous wing pitch and camber are controlled automatically, rather than by muscular action; probably in association with the development of asynchronous power musculature.
Tillyard's Cu2 (=CUP) is believed to be a secondary pseudo-vein, his 1A to be the true CuP and 2A to be 1A.
The late Permian fossil Permotipula Patricia is almost certainly a member of the Diptera stem-group, possibly even of the crown-group. The Mesozoic Laurentipteridae and the Permian Permotanyderidae are other possible, but not certain, stem-group members.     

Locomotor Behavior of Flightless Harmonia axyridis Pallas (Col., Coccinellidae)

To improve the efficiency of the lady beetle H. axyridis as a biological control agent against aphids, a flightless population was obtained by feeding adults with a mutagen and selecting their progeny for nonflying but otherwise morphologically normal individuals. These flightless adults attempted to fly but immediately fell. They softened their fall by opening their elytrae and wings. The inability to fly could result from change in their flying behavior compared to control adults. The flight duration was very much shorter, and the wing beat frequency and, more particularly, the amplitude of the wing beats were clearly lower. More time was spent in the other components of the flight behavior such as wing rotation, wing immobility, and wing folding. The sequence of these patterns differed slightly, due mainly to change in their frequency. The locomotor behavior was not modified by the mutation, which affected only the wing muscles. Searching behavior of mutant adults differed from that of control adults only in that they took longer to encounter and ingest aphids. Nevertheless, the larval growth and reproductive rate remained unchanged. The behavioral and biological features of these flightless adults indicate that it should be possible to use them in biological control programs.     

Designing a Biomimetic Ornithopter Capable of Sustained and Controlled Flight

We describe the design of four ornithopters ranging in wing span from 10 cm to 40 cm, and in weight from 5 g to 45 g. The controllability and power supply are two major considerations, so we compare the efficiency and characteristics between different types of subsystems such as gearbox and tail shape. Our current omithopter is radio-controlled with inbuilt visual sensing and capable of takeoff and landing. We also concentrate on its wing efficiency based on design inspired by a real insect wing and consider that aspects of insect flight such as delayed stall and wake capture are essential at such small size. Most importantly, the advance ratio, controlled either by enlarging the wing beat amplitude or raising the wing beat frequency, is the most significant factor in an ornithopter which mimics an insect.     

Locust flight initiation: a comparison of normal and artificial release

Flight initiation was analysed in the migratory locust, Locusta migratoria L. using conventional and high‐speed video systems. Flight was initiated by three methods: normal jump, free fall and controlled catapult. Parameters evaluated were time from release to wing‐opening, body angle, speed, height gain, wing‐opening, initial wing‐beat frequency. Fastest wing‐opening occurred following a normal jump. A catapult device allowed manipulation of the launching parameters: speed and angles at launching. It appeared that in an artificially catapulted start there was a minimum speed (v > 0.75 m/s) required to initiate flight. However, under free‐fall conditions a mean speed of v = 0.6 m/s at wing‐opening was observed. When the different parameters of the controlled catapult start were equal to those for normal jump then the time to wing‐opening was found to be extended for the catapult launch. However, other parameters were not affected, occasionally even a ‘kick in air’ was observed. The catapult launches indicated that within about three wing‐beat cycles the animals showed active flight, with positive lift and constant or increasing speed, compared to a ballistic trajectory. Our results indicate that a controlled catapult device will prove useful to the study of sensory and central processes underlying free flight initiation.     

Direct measurements of the kinematics and dynamics of bat flight

Experimental measurements and analysis of the flight of bats are presented, including kinematic analysis of high-speed stereo videography of straight and turning flight, and measurements of the wake velocity field behind the bat. The kinematic data reveal that, at relatively slow flight speeds, wing motion is quite complex, including a sharp retraction of the wing during the upstroke and a broad sweep of the partially extended wing during the downstroke. The data also indicate that the flight speed and elevation are not constant, but oscillate in synchrony with both the horizontal and vertical movements of the wing. PIV measurements in the transverse (Trefftz) plane of the wake indicate a complex 'wake vortex' structure dominated by a strong wing tip vortex shed from the wing tip during the downstroke and either the wing tip or a more proximal joint during the upstroke. Data synthesis of several discrete realizations suggests a 'cartoon' of the wake structure during the entire wing beat cycle. Considerable work remains to be done to confirm and amplify these results.