Structural and functional development of cricket ring muscles

The sizes of the unifunctional dorsal longitudinal (DLM) and bifunctional subalar (SA) metathoracic flight muscles of the cricket Teleogryllus oceanicus increase by more than an order of magnitude between the second instar before the terminal molt and the tenth day of adult life. During the same developmental period isometric twitch duration (onset to 50% relaxation, 25 degrees C) varies little, while muscle mitochondrial content increased by a factor of ten as measured by stereological analysis of electron micrographs and citrate synthase activity (mumoles citrate . min-1 . gm protein-1, 25 degrees C). The wing muscles of adults have abundant sarcoplasmic reticulum (SR), narrow myofibrils, and a high volume density of mitochondria. At two molts from adulthood muscles that will later be used in flight behavior also have narrow myofibrils and abundant SR, but unlike muscles at later stages, nymphal muscles have a low volume density of mitochondria. At the terminal molt muscles have at least as much SR as is seen in muscles at the tenth day of adult life, and the myofibrils are also more narrow at the earlier stage. Since there is significant variation in muscle structure and little change in twitch duration during late development, the efficacy of the SR in releasing and resequestering CA2+ is seemingly lower in muscles at the terminal molt, a time of rapid muscle growth.     

Innervation is necessary for the development of fast contraction kinetics of singing muscles in a katydid

The twitch duration of mesothoracic wing muscles of the male katydid Neoconocephalus robustus (Insecta; Orthoptera; Tettigoniidae) decreases rapidly within the first 5 days of adulthood, to about half of its value in newly molted adults. To determine if this change is dependent upon neural input, male mesothoracic first tergocoxal muscles were unilaterally denervated on the second day of adulthood. The contraction kinetics of the denervated and contralateral innervated muscles were tested four days later. The development of rapid contraction kinetics was slowed or stopped in the denervated muscles, while the contralateral innervated muscles did become faster. Mesothoracic wing muscles of females do not develop faster contraction kinetics. When the female mesothoracic first tergocoxal muscle is denervated, there is no difference in twitch duration after 4 days between the innervated and contralateral denervated muscles. Therefore, denervation in newly molted adult male katydids interrupts a developmental program for the acquisition of adult contraction kinetics.     

Comparative morphology of the thorax musculature of adult Anisoptera (Insecta: Odonata): Functional aspects of the flight apparatus

Due to their unique flight mechanism including a direct flight musculature, Odonata show impressive flight skills. Several publications addressed the details of this flight apparatus like: sclerites, wings, musculature, and flight aerodynamics. However, 3D-analysis of the thorax musculature of adult dragonflies was not studied before and this paper allows for a detailed insight. We, therefore, focused on the thorax musculature of adult Anisoptera using micro-computed tomography. Herewith, we present a comparative morphological approach to identify differences within Anisoptera: Aeshnidae, Corduliidae, Gomphidae, and Libellulidae. In total, 54 muscles were identified: 16 prothoracic, 19 mesothoracic, and 19 metathoracic. Recorded differences were for example, the reduction of muscle Idlm4 and an additional muscle IIIdlm1 in Aeshna cyanea, previously described as rudimentary or missing. Muscle Iscm1, which was previously reported missing in all Odonata, was found in all investigated species. The attachment of muscle IIpcm2 in Pantala flavescens is interpreted as a probable adaption to its long-distance migration behaviour. Furthermore, we present a review of functions of the odonatan flight muscles, considering previous publications. The data herein set a basis for functional and biomechanical studies of the flight apparatus and will therefore lay the foundation for a better understanding of the odonatan flight.     

Monosynaptic connexions between wing stretch receptors and flight motoneurones of the locust.

1. The connexions between stretch receptors of the wings and motoneurones innervating flight muscles have been studied anatomically and physiologically. 2. Filling with cobaltous chloride shows that the single neurone of a forewing stretch receptor has a complex pattern of branches within the mesothoracic ganglion and branches which extend into the pro- and meta-thoracic ganglia. The single neurone of a hindwing stretch receptor has extensive branches in the metathoracic ganglion and branches in themesothoracic ganglion. The branches of both receptors are confined to the ipsilateral halves of the ganglia. 3. A stretch receptor gives information about the velocity and extent of elevation of a wing. 4. Each spike of a forewing stretch receptor casuses an EPSP in ipsilateral mesothoracic depressor motoneurones and an IPSP in elevators. The connexions are thought to be monosynaptic for the following reasons. The EPSPs in the first basalar (depressor) motoneurone follow each spike of the stretch receptor at a frequency of 125 Hz and with a constant latency of about 1 msec. In a Ringer solution containing 20 mM-Mg2+ the amplitude EPSP declines gradually. The IPSP'S upon elevators have similar properties but occur with a latency of 4-6 msec. 5. The connexions therefore comprise a monosynaptic negative feed-back loop; elevation of the wing excites the stretch receptor which then inhibits the elevator motoneurones and excites the depressors. 6. A hindwing stretch receptor synapses upon metathoracic flight motoneurones in the same way, causing EPSPs in depressor and IPSPs in elevator motoneurones. 7. No connexions of either fore- or hindwing stretch receptors have been found with contralateral flight motoneurones. 8. Interganglionic connexions are made by both receptors. For example, both fore- and hindwing stretch receptors cause EPSPs upon the meso- and metathoracic first basalar motoneurones. 9. Stimulation of the axon of a stretch receptor with groups of three stimuli repeated every 50-100 msec thus simulating the pattern which it shows during flight, causes subthreshold waves of depolarization in depressor motoneurones. When summed with an unpatterned input, the stretch receptor is able to influence the production of spikes in motoneurones on each cycle. During flight, it is expected that the stretch receptor will influence the time at which a motoneurone will spike and hence have an effect on the amplitude of the upstroke and upon the phase relationship between spikes of motoneurones.     

Steering responses of adult and nymphal crickets to light,with special reference to the head rolling movement

Most tethered adult crickets (Gryllus bimaculatus) assumed flight postures with or without flapping their wings in a windstream. Nymphal crickets (sixth and seventh, i.e. final, instars) also displayed the flight posture in spite of the incompleteness of wing development. These adult nymphal crickets rolled their heads towards the light source in response to unequal illumination of the compound eyes only while maintaining the flight posture. The amphtude of the head rolling movements was proportional to the change of light position up to 120°C, and independent of the light intensity if the duration was longer than 1 sec. The unequal illumination could also induce a transient increase in discharge frequency of the wing muscles on both sides, a decrease in wing beat amplitude of the ipsilateral wing on the illuminated side, and bending movements of the legs and abdomen towards the light. Cutting either of the nerve connectives at any level between the subosophageal and metathoracic ganglia did not affect the response of either the head or the abdomen to illumination. These results are discussed in relation to the steering mechanism associated with the dorsal light reaction.     

Rapid postembryonic development of a cricket flight muscle

The metathoracic dorsal longitudinal muscle (DLM) of the cricket Teleogryllus oceanicus differentiated and developed rapidly over the last nymphal instar. Within eight days, the muscle mass increased by a factor of 15 and the relative volume of mitochondria quadrupled, while the relative amounts of myofibril and sarcoplasmic reticulum decreased. Tracheoblasts began to invade the muscle fibers immediately before the adult molt. Muscle mass continued to increase until four days after the adult molt, but the relative volumes of the ultrastructural components did not change. Within two weeks following the adult molt, the muscles in some of the animals began to degenerate.     

Central projections of the wing afferents in the hawkmoth, Agrius convolvuli

Flight behaviors in various insect species are closely correlated with their mechanical and neuronal properties. Compared to locusts and flies which have been intensively studied, moths have “intermediate” properties in terms of the neurogenic muscle activations, power generation by indirect muscles, and two-winged-insect-like flapping behavior. Despite these unique characteristics, little is known about the neuronal mechanisms of flight control in moths. We investigated projections of the wing mechanosensory afferents in the central nervous system (CNS) of the hawkmoth, Agrius convolvuli, because the mechanosensory proprioceptive feedback has an essential role for flight control and would be presumably optimized for insect species. We conducted anterograde staining of nine afferent nerves from the fore- and hindwings. All of these afferents projected into the prothoracic, mesothoracic and metathoracic ganglia (TG1, 2 and 3) and had ascending fibers to the head ganglia. Prominent projection areas in the TG1–3 and suboesophageal ganglion (SOG) were common between the forewing, hindwing and contralateral forewing afferents, suggesting that information from different wings are converged at multiple levels presumably for coordinating wing flapping. On the other hand, differences of projections between the fore- and hindwing afferents were observed especially in projection areas of the tegulae in the TG1 and contralateral projections of the anterior forewing nerve in the TGs and SOG, which would reflect functional differences between corresponding mechanoreceptors on each wing. Afferents comprising groups of the campaniform sensilla at the wing bases had prominent ascending pathways to the SOG, resembling the head–neck motor system for gaze control in flies. Double staining of the wing afferents and flight or neck motoneurons also indicated potential connectivity between them. Our results suggest multiple roles of the wing proprioceptive feedback for flight and provide the anatomical basis for further understanding of neuronal mechanisms of the flight system in moths.     

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" (or "startle") response during the adult stage is mediated by the multi-component neuronal circuit called the Giant Fiber System (GFS). The comparative large size of the neurons, their distinctive morphology and simple connectivity make the GFS an attractive model system for studying neuronal circuitry. The GFS pathway is composed of two bilaterally symmetrical Giant Fiber (GF) interneurons whose axons descend from the brain along the midline into the thoracic ganglion via the cervical connective. In the mesothoracic neuromere (T2) of the ventral ganglia the GFs form electro-chemical synapses with 1) the large medial dendrite of the ipsilateral motorneuron (TTMn) which drives the tergotrochanteral muscle (TTM), the main extensor for the mesothoracic femur/leg, and 2) the contralateral peripherally synapsing interneuron (PSI) which in turn forms chemical (cholinergic) synapses with the motorneurons (DLMns) of the dorsal longitudinal muscles (DLMs), the wing depressors. The neuronal pathway(s) to the dorsovental muscles (DVMs), the wing elevators, has not yet been worked out (the DLMs and DVMs are known jointly as indirect flight muscles - they are not attached directly to the wings, but rather move the wings indirectly by distorting the nearby thoracic cuticle) (King and Wyman, 1980; Allen et al., 2006). The di-synaptic activation of the DLMs (via PSI) causes a small but important delay in the timing of the contraction of these muscles relative to the monosynaptic activation of TTM (~0.5 ms) allowing the TTMs to first extend the femur and propel the fly off the ground. The TTMs simultaneously stretch-activate the DLMs which in turn mutually stretch-activate the DVMs for the duration of the flight. The GF pathway can be activated either indirectly by applying a sensory (e.g."air-puff" or "lights-off") stimulus, or directly by a supra-threshold electrical stimulus to the brain (described here). In both cases, an action potential reaches the TTMs and DLMs solely via the GFs, PSIs, and TTM/DLM motoneurons, although the TTMns and DLMns do have other, as yet unidentified, sensory inputs. Measuring "latency response" (the time between the stimulation and muscle depolarization) and the "following to high frequency stimulation" (the number of successful responses to a certain number of high frequency stimuli) provides a way to reproducibly and quantitatively assess the functional status of the GFS components, including both central synapses (GF-TTMn, GF-PSI, PSI-DLMn) and the chemical (glutamatergic) neuromuscular junctions (TTMn-TTM and DLMn-DLM). It has been used to identify genes involved in central synapse formation and to assess CNS function.     

Modulation of tension production by proctolin in the dorsal longitudinal muscles of the cricket,Teleogryllus oceanicus

High-frequency electrical stimulation (～20 Hz) of the lateral nerve in abdominal segments of the cricket, Teleogryllus oceanicus, caused an increase in tonus of the abdominal dorsal longitudinal muscle (DLM). This effect persisted for 1–5 min following stimulation. Application of the pentapeptide proctolin (threshold 1–10 nM) mimicked the increase in muscle tonus produced by electrical stimulation. Individual twitches were unaffected or slightly reduced by proctolin. Low-frequency electrical stimulation (<7 Hz) of the lateral nerve counteracted a previously induced increase in muscle tonus, apparently by activation of an inhibitory motoneuron. γ-Aminobutyric acid (GABA) mimicked the effect of low-frequency stimulation and reduced muscle tonus. Octopamine, in concentrations of ≤0.1 mM, was inactive on the abdominal DLM when stimulated at low frequencies (0.5–2 Hz). Application of proctolin to the metathoracic DLM caused an increase in twitch amplitude but had little effect on basal tonus. In conjunction with the previously described responses of the metathoracic DLM to octopamine, these results show that the serially homologous abdominal and metathoracic DLMs have dissimilar responses to the modulators proctolin and octopamine.     

Control of growth and ultrastructural maturation of a cricket flight muscle

The metathoracic dorsal longitudinal muscle (DLM) in the cricket Teleogryllus oceanicus increased in mass and rapidly acquired interfibrillar tracheoles and an increased proportion of mitochondria around the time of the adult molt. Both neural input and juvenile hormone levels were investigated as possible factors controlling this rapid maturation. Motor axons to the muscle were cut early in the last nymphal instar, and muscle growth slowed but ultrastructural maturation continued; the percentage of muscle volume occupied by mitochondria tripled and tracheoblasts invaded the fibers in both the denervated and contralateral innervated muscles. Newly molted last instar nymphs were treated with methoprene, a juvenile hormone analog, and examined four days following the next molt. Muscle growth was slowed but not stopped. Both mitochondrial proliferation and tracheoblast formation were completely blocked by hormone treatment. This study shows that both neural input and low levels of juvenile hormone are required for muscle growth. However, ultrastructural maturation seems to depend exclusively on low levels of juvenile hormone.     

Developmental analysis of some mutants of the bithorax system ofDrosophila

Summary Mutants in the bithorax system ofDrosophila produce homeotic transformations that affect the mesothoracic, metathoracic and first abdominal segments. In the present report we describe a clonal analysis of the development of those mutants transforming the metathorax and first abdominal segments into mesothorax.The main results indicate that (1) The normal dorsal metathoracic (haltere) disk has similar developmental parameters to the dorsal mesothoracic disk. The main difference is that the initial and final numbers of cells are different in both disks. (2) In flies mutant forBithorax andpostbithorax (which transform the haltere into wing) the transformed haltere disk has the same initial and final number of cells as the normal wing disk. (3) In morphogenetic mosaics homozygousbithorax (andpostbithorax) clones express their genotype autonomously regardless of the genotype of surrounding haltere cells. This autonomy is expressed in a regulation of the number of adult cells per compartment, typical cell affinities and final cuticular differentiation.     

Chronic stimulation modifies the isotonic shortening velocity of denervated rat slow-twitch muscle

Rat soleus muscles were denervated and stimulated in vivo for periods of up to 104 days. Stimuli used were trains of 1 ms pulses at 100 Hz delivered for periods of 1 s; trains were repeated every 10-100 s. In a majority of animals the tension of the muscles was maintained at about 10% of normal, equivalent to muscles denervated but unstimulated for 20 days. At the longest periods the stimulated muscles developed ten times more tension than ones that were denervated but not stimulated. In denervated and denervated-stimulated muscles twitch contraction and relaxation times were prolonged, compared with controls, for up to 3 weeks. Thereafter both sets showed a speeding of the isometric twitch that was greater in the stimulated muscles. At the longest periods the twitch was as short as that of a denervated fast muscle. Stimulation did not affect contralateral denervated muscles. Twitch: tetanus ratios remained high despite stimulation, and muscles showed little post-tetanic potentiation. Tension developed more rapidly in the tetani of the stimulated muscles, even allowing for larger final values. Maximum velocity of shortening was increased in many of the stimulated muscles, and there was a proportional flattening of the force-velocity curve, i.e. a/P0 increased. Maximum velocity and a/P0 increased reciprocally with twitch time to peak, so that those muscles that had twitches most changed by stimulation also had their isotonic properties modified to the greatest extent. Even at the longest period of stimulation, twitch time course and tetanic tension were not converted to those of normal fast muscle.     

Locust flight behavior after hemisection of individual thoracic ganglia: evidence for hemiganglionic premotor centers

Summary The flight behavior of locusts with hemisected mesothoracic or metathoracic ganglia was observed in unrestrained animals and monitored electromyographically in tethered animals. Animals with hemisected mesothoracic ganglia were able to initiate and carry out free flight. Hemisection of the mesothoracic ganglion caused no significant changes in the pattern of flight muscle firing; both intra- and intersegmental coordination of flight muscle activity were retained (Figs. 3, 4). Additional transection of one meso-metathoracic connective altered the pattern of flight muscle firing but did not abolish rhythmic activity (Fig. 8). Deafferentation of the thoracic ganglia in animals with hemisected mesothoracic ganglia resulted in rhythmically coordinated motor activity (Fig. 5) which was indistinguishable from that shown by deafferented animals with all ganglia intact. Hemisection of the metathoracic ganglion resulted in an abnormal pattern of flight muscle firing. However, a basic rhythmicity of motor activity was still present (Fig. 6). The implications of these results for rhythm generation and motor coordination in the flight control system of the locust are discussed.     

Output connections of a wind sensitive interneurone with motor neurones innervating flight steering muscles in the locust

Summary The output connections of a bilaterally symmetrical pair of wind-sensitive interneurones (called A4I1) were determined in a non-flying locust (Schistocerca gregaria). Direct inputs from sensory neurones of specific prosternai and head hairs initiate spikes in these interneurones in the prothoracic ganglion.The interneurone with its axon in the right connective makes direct, excitatory connections with the two mesothoracic motor neurones innervating the pleuroaxillary (pleuroalar, M85) muscle of the right forewing, but not with the comparable motor neurones of the left forewing. The connections can evoke motor spikes.The interneurones also exert a powerful, but indirect effect on the homologous metathoracic pleuroaxillary motor neurones (muscle 114), and a weaker, indirect effect on subalar motor neurones of the hindwings. No connections or effects were found with other flight motor neurones, or motor neurones innervating hindleg muscles, including common inhibitor 1 which also innervates the pleuroaxillary muscle.One thoracic interneurone with its cell body in the right half of the mesothoracic ganglion and with its axon projecting ipsilaterally to the metathoracic ganglion receives a direct input from the right A4I1 interneurone.These restricted output connections suggest a role for the A4I1 interneurones in flight steering.Abbreviations DCMD descending contralateral movement detector - EPSP excitatory postsynaptic potential - TCG tritocerebral commissure giant (interneurone)     

Ultrabithorax gene expression in Drosophila imaginal discs and larval nervous system

Using a monoclonal antibody and image-processing procedures, the patterns of expression of the Ultrabithorax (Ubx) gene product have been characterized in Drosophila larvae. As reported previously, the metathoracic imaginal discs stain most intensely with anti-Ubx, with some mesothoracic and no prothoracic expression detectable. In the metathoracic discs, the greatest modulation in anti-Ubx staining is along the proximodistal axis. Ubx is generally expressed at higher levels in the posterior regions of metathoracic discs, although relatively high anterior expression is found in some areas. Expression in the mature wing disc is confined to the squamous peripodial membrane cells; in younger wings, Ubx expression fills the posterior half of the peripodial side of the disc. The mesothoracic leg stains with a pattern that is qualitatively similar (but not identical) to that of the metathoracic leg; Ubx is expressed in some anterior regions of the mesothoracic leg, in parasegment 4. Double staining with anti-Ubx and anti-engrailed reveals that discontinuities in Ubx expression that have been suggested to correspond to compartment borders do not coincide with the compartment boundaries in some cases. In the larval ventral ganglion, Ubx expression is greatest in parasegments 5 and 6, as in the embryonic nervous system.     

Juvenile hormone titer and morph-specific reproduction in the wing-polymorphic cricket,Gryllus firmus

Juvenile hormone titers and reproductive characteristics were measured in adult wing and flight-muscle morphs of the wing-polymorphic cricket, Gryllus firmus, during the first week of adulthood. This species has three morphs: one flight capable morph with fully-developed wings and fully-developed flight muscles [LW(F)], one flightless morph with fully-developed wings and histolyzed (non-functional) flight muscles [LW(H)], and another flightless morph with underdeveloped (short) wings and underdeveloped flight muscles (SW). Both flightless morphs [LW(H) and SW] had larger ovaries which contained a greater number of postvitellogenic eggs compared with the flight capable [LW(F)] morph. The juvenile hormone titer was significantly higher in SW compared with LW(F) females on days 3-7 of adulthood. On these days, the JH titer also was significantly higher in the other flightless morph, LW(H), compared with flight-capable [LW(F)] females as determined by one statistical test, but did not differ significantly by another test. The JH titer was positively correlated with ovarian mass or terminal oocyte length, but not with the number of post-vitellogenic eggs. This study is the first direct comparison of juvenile hormone titers in adult wing morphs of a wing-polymorphic insect. Results indicate that an elevated juvenile hormone titer may be at least partly responsible for one of the most distinctive features of wing-polymorphic species, the increased early fecundity of flightless females.     

Phylogenetic Relationships of the Suborders of Coleoptera (Insecta)

One hundred seven external and internal characters of larval and adult representatives of 28 genera of the coleopteran suborders were analyzed cladistically. Four groups of Neuropterida were introduced as outgroup. The analysis yielded 18 trees with a minimum of 194 steps (CI 0.691). All trees support the monophyly of all four suborders and a branching pattern (Archostemata + (Adephaga + (Myxophaga + Polyphaga))). The presence of elytra with meso- and metathoracic locking devices, the specific hind-wing folding, the close connection of exposed sclerites, the absence of the mera, the absence of eight thoracic muscles, the reduced abdominal sternite I, and the invagination of terminal segments are autapomorphies of Coleoptera. The monophyly of Coleoptera excl. Archostemata is supported by further transformations of the thoracic sclerites such as absence of the mesothoracic discriminal line and katepisternal joint, by an internalized or absent metathoracic trochantin, by the presence of a bending zone in the hind-wing, and by eight further muscle losses. Fusion of tibia and tarsus and presence of a single claw are larval synapomorphies of Myxophaga and Polyphaga. Adults are characterized by fusion of protrochantin and propleura and by the rigid connection of the meso- and metathoracic ventrites. The eucinetoid lineage of Polyphaga is characterized by the secondary absence of the bending zone of the alae. This results in a distinctly simplified wing folding mechanism. The monophyly of Cucujiformia (+ Bostrichoidea) is supported by the presence of cryptonephric Malpighian tubules. Transformations of fore-and hind-wings, reinforcement and simplification of the thoracic exoskeleton, and an efficient use of a distinctly reduced set of thoracic muscles play an important role in the early evolution of Coleoptera. Many different larval character transformations take place in the earlier Mesozoic within the suborders.     

Flight muscles polymorphism in a flightless bug, Pyrrhocoris apterus (L.): developmental pattern, biochemical profile and endocrine control

The flightless bug Pyrrhocoris apterus (L.) is polymorphic for both wing length and flight muscle development. The developed flight muscles of macropterous adults of both sexes first enlarge their volume during the first 5 days after adult emergence, but are then histolyzed in all males and females older than 10 and 14 days, respectively. The flight muscles of brachypterous adult males and females are underdeveloped due to their arrested growth. The total protein content of histolyzed dorsolongitudinal flight muscles from 21-day-old macropterous adults of both sexes is lower than that of developed dorsolongitudinal flight muscles in 5-10-days-old macropterous bugs, but substantially higher than the protein content of underdeveloped dorsolongitudinal flight muscles from adult brachypters. Histolyzed dorsolongitudinal flight muscles differ from the developed ones by decreased quantities of 18 electrophoretically separated proteins. Histolysis of developed dorsolongitudinal flight muscles is accompanied by significant decreases in citrate synthase, glyceraldehyde-3-phosphate dehydrogenase and β-hydroxyacyl-CoA dehydrogenase enzyme activities and an increase in alanine aminotransferase activity, and can be precociously induced by application of a juvenile hormone analogue. This is the first report of flight muscle polymorphism, histolysis of developed flight muscles and its endocrine control in insects displaying non-functional wing polymorphism.     

Structural and functional heterogeneity in an insect muscle.

Singing muscles of the katydid, Neoconocephalus robustus (Insecta, Tettigoniidae) are neurogenic, yet perform at contraction-relaxation frequencies as high as 212 Hz (Josephson and Halverson, '71). The mechanical and electrical responses of different bands of one of these muscles (the dorsal longitudinal muscle, DLM) has been examined with respect to ultrastructural features of each part which may be related to muscle performance. The DLM is composed of three bands and is innervated by four motoneurones. The cell bodies of three of these motoneurones occur ipsilaterally in the prothroracic ganglion; the cell body of the other motoneurone is contralateral in the mesothoracic ganglion. Three of the motoneurones (as yet unidentified fast axons) initiate extraordinarily fast twitches (rise time equal 7.3 msec, half duration equals 14.3 msec, 25 C), the fourth (an unidentified slower axon) evokes twitches which are considerably slower (rise time equals 18.9 msec, half duration equals 5.10 msec). Whereas the ventral and medial bands of the muscle are innervated only by fast axons (some fibers of the medial band are doubly innervated), the dorsal band is innervated by both a fast axon and the slower axon. A few fibers of the dorsal band are doubly innervated. The structure of fibers from the ventral and medial bands is very similar, with short sarcomeres (4.0 and 4.3 mum, respectively) and thin strap-like myofibrils delineated by well-developed sarcoplasmic reticulum (SR). Twenty-four percent of the volume of ventral band fibers is SR and the diffusion distance from SR to the center of the adjacent myofibril averages 0.083 mum. Twenty percent of the medial band fiber volume is SR, with a diffusion distance of 0.118 mum. Ventral and medial band fibers contain about 40% mitochondria, and 33% myofibrils. The dorsal band fibers have longer sarcomeres (9.5 mum), and only 10% of the fiber volume is SR. The muscle fibrils of the dorsal band are larger and consequently the diffusion distance is greater (0.227 mum) than in the ventral and medial bands. Mitochondria comprise 23% of the volume of dorsal band fibers. Most dorsal band mitochondria are aggregated into distinct clumps. Although some dorsal band fibers are innervated by a fast axon and some by the slower axon, the dorsal band fibers are structurally homogeneous, suggesting that neurotrophic effects are not important in maintaining the structure of dorsal band fibers. The mechanical-electrical performance and ultrastructure of the ventral and medial bands suggest their roll as fast, metabolically active but weak muscles, used in singing; the dorsal band as a slower but stronger muscle, perhaps involved in postural movements of the wing during singing.