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Developmental constraint of insect audition.

BACKGROUND: Insect ears contain very different numbers of sensory cells, from only one sensory cell in some moths to thousands of sensory cells, e.g. in cicadas. These differences still await functional explanation and especially the large numbers in cicadas remain puzzling. Insects of the different orders have distinct developmental sequences for the generation of auditory organs. These sensory cells might have different functions depending on the developmental stages. Here we propose that constraints arising during development are also important for the design of insect ears and might influence cell numbers of the adults. PRESENTATION OF THE HYPOTHESIS: We propose that the functional requirements of the subadult stages determine the adult complement of sensory units in the auditory system of cicadas. The hypothetical larval sensory organ should function as a vibration receiver, representing a functional caenogenesis. TESTING THE HYPOTHESIS: Experiments at different levels have to be designed to test the hypothesis. Firstly, the neuroanatomy of the larval sense organ should be analyzed to detail. Secondly, the function should be unraveled neurophysiologically and behaviorally. Thirdly, the persistence of the sensory cells and the rebuilding of the sensory organ to the adult should be investigated. IMPLICATIONS OF THE HYPOTHESIS: Usually, the evolution of insect ears is viewed with respect to physiological and neuronal mechanisms of sound perception. This view should be extended to the development of sense organs. Functional requirements during postembryonic development may act as constraints for the evolution of adult organs, as exemplified with the auditory system of cicadas.

Journal Article↗

Threatened edible insects in Hidalgo, Mexico and some measures to preserve them.

Edible insects are a natural renewable resource that provides food to many ethnic groups in Mexico. Some of these species are overexploited because of increased consumption, caused by the huge human population growth in the area and because of the large demand of these insects from many restaurants in Mexico and in other countries. In Tulancalco, a small arid village in the State of Hidalgo, I carried out studies on edible insects over 25 years. The inhabitants of this village have a natural economy and use some 30 species of insects as food. At present, we have noticed a decrease in the population of several species due to overexploitation, which is carried by non-qualified independent workers who are not natives of the town. These gatherers sell their catch to make a living, thus contributing to the socioeconomic factors associated with this issue. These actions have degraded the ecosystems of this area, and consequently the prevention of these measures is critical. The study species in this paper include 14 threatened species and we discuss some pragmatic measures that could implemented to avoid their extinction. In addition, some actions for the preservation of the ethnoentomobiodiversity in the area are proposed.

Agriculture↗

Biology of Toll receptors: lessons from insects and mammals.

Toll receptors are type I transmembrane proteins that play important roles in development and immunity in animals. Comparison of the genomes of mouse and human on one side and of the fruitfly Drosophila and the mosquito Anopheles (two dipteran insects) on the other, revealed that the four species possess a similar number of Toll receptors (approximately 10). However, phylogenetic analyses indicate that the families of Toll receptors expanded independently in insects and mammals. We review recent results on these receptors, which point to differences in the activation and signaling between Tolls in insects and Toll-like receptors (TLRs) in mammals. Whereas mammalian TLRs appear to be solely dedicated to host-defense, insect Tolls may be predominantly linked to other functions, probably developmental.

Animals↗

Kruppel is a gap gene in the intermediate germband insect Oncopeltus fasciatus and is required for development of both blastoderm and germband-derived segments.

Segmentation in long germband insects such as Drosophila occurs essentially simultaneously across the entire body. A cascade of segmentation genes patterns the embryo along its anterior-posterior axis via subdivision of the blastoderm. This is in contrast to short and intermediate germband modes of segmentation where the anterior segments are formed during the blastoderm stage and the remaining posterior segments arise at later stages from a posterior growth zone. The biphasic character of segment generation in short and intermediate germ insects implies that different formative mechanisms may be operating in blastoderm-derived and germband-derived segments. In Drosophila, the gap gene Krüppel is required for proper formation of the central portion of the embryo. This domain of Krüppel activity in Drosophila corresponds to a region that in short and intermediate germband insects spans both blastoderm and germband-derived segments. We have cloned the Krüppel homolog from the milkweed bug, Oncopeltus fasciatus (Hemiptera, Lygaeidae), an intermediate germband insect. We find that Oncopeltus Krüppel is expressed in a gap-like domain in the thorax during the blastoderm and germband stages of embryogenesis. In order to investigate the function of Krüppel in Oncopeltus segmentation, we generated knockdown phenotypes using RNAi. Loss of Krüppel activity in Oncopeltus results in a large gap phenotype, with loss of the mesothoracic through fourth abdominal segments. Additionally, we find that Krüppel is required to suppress both anterior and posterior Hox gene expression in the central portion of the germband. Our results show that Krüppel is required for both blastoderm-derived and germband-derived segments and indicate that Krüppel function is largely conserved in Oncopeltus and Drosophila despite their divergent embryogenesis.

Amino Acid Sequence↗

Flexural stiffness in insect wings. I. Scaling and the influence of wing venation.

During flight, many insect wings undergo dramatic deformations that are controlled largely by the architecture of the wing. The pattern of supporting veins in wings varies widely among insect orders and families, but the functional significance of phylogenetic trends in wing venation remains unknown, and measurements of the mechanical properties of wings are rare. In this study, we address the relationship between venation pattern and wing flexibility by measuring the flexural stiffness of wings (in both the spanwise and chordwise directions) and quantifying wing venation in 16 insect species from six orders. These measurements show that spanwise flexural stiffness scales strongly with the cube of wing span, whereas chordwise flexural stiffness scales with the square of chord length. Wing size accounts for over 95% of the variability in measured flexural stiffness; the residuals of this relationship are small and uncorrelated with standardized independent contrasts of wing venation characters. In all species tested, spanwise flexural stiffness is 1-2 orders of magnitude larger than chordwise flexural stiffness. A finite element model of an insect wing demonstrates that leading edge veins are crucial in generating this spanwise-chordwise anisotropy.

Animals↗

Flexural stiffness in insect wings. II. Spatial distribution and dynamic wing bending.

The dynamic, three-dimensional shape of flapping insect wings may influence many aspects of flight performance. Insect wing deformations during flight are largely passive, and are controlled primarily by the architecture and material properties of the wing. Although many details of wing structure are well understood, the distribution of flexural stiffness in insect wings and its effects on wing bending are unknown. In this study, we developed a method of estimating spatial variation in flexural stiffness in both the spanwise and chordwise direction of insect wings. We measured displacement along the wing in response to a point force, and modeled flexural stiffness variation as a simple mathematical function capable of approximating this measured displacement. We used this method to estimate flexural stiffness variation in the hawkmoth Manduca sexta, and the dragonfly Aeshna multicolor. In both species, flexural stiffness declines sharply from the wing base to the tip, and from the leading edge to the trailing edge; this variation can be approximated by an exponential decline. The wings of M. sexta also display dorsal/ventral asymmetry in flexural stiffness and significant differences between males and females. Finite element models based on M. sexta forewings demonstrate that the measured spatial variation in flexural stiffness preserves rigidity in proximal regions of the wing, while transferring bending to the edges, where aerodynamic force production is most sensitive to subtle changes in shape.

Animals↗

Chitin metabolism in insects: structure, function and regulation of chitin synthases and chitinases.

Chitin is one of the most important biopolymers in nature. It is mainly produced by fungi, arthropods and nematodes. In insects, it functions as scaffold material, supporting the cuticles of the epidermis and trachea as well as the peritrophic matrices lining the gut epithelium. Insect growth and morphogenesis are strictly dependent on the capability to remodel chitin-containing structures. For this purpose, insects repeatedly produce chitin synthases and chitinolytic enzymes in different tissues. Coordination of chitin synthesis and its degradation requires strict control of the participating enzymes during development. In this review, we will summarize recent advances in understanding chitin synthesis and its degradation in insects.

Animals↗

The fluid dynamics of flight control by kinematic phase lag variation between two robotic insect wings.

Insects flying with two pairs of wings must contend with the forewing wake passing over the beating hindwing. Some four-winged insects, such as dragonflies, move each wing independently and therefore may alter the relative timing between the fore- and hindwing stroke cycles. The significance of modifying the phase relationship between fore- and hindwing stroke kinematics on total lift production is difficult to assess in the flying animal because the effect of wing-wake interference critically depends on the complex wake pattern produced by the two beating wings. Here we investigate the effect of changing the fore- and hindwing stroke-phase relationship during hovering flight conditions on the aerodynamic performance of each flapping wing by using a dynamically scaled electromechanical insect model. By varying the relative phase difference between fore- and hindwing stroke cycles we found that the performance of the forewing remains approximately constant, while hindwing lift production may vary by a factor of two. Hindwing lift modulation appears to be due to two different fluid dynamic phenomenons: leading edge vortex destruction and changes in strength and orientation of the local flow vector. Unexpectedly, the hindwing regains aerodynamic performance near to that of the wing free from forewing wake interference, when the motion of the hindwing leads the forewing by around a quarter of the stroke cycle. This kinematic relationship between hind- and forewing closely matches the phase-shift commonly used by locusts and some dragonflies in climbing and forward flight. The experiments support previous assumptions that active neuromuscular control of fore- and hindwing stroke phase might enable dragonflies and other functionally four-winged insects to manipulate ipsilateral flight force production without further changes in wing beat kinematics.

Animals↗

Induced airflow in flying insects I. A theoretical model of the induced flow.

A strong induced flow structure envelops the body of insects and birds during flight. This flow influences many physiological processes including delivery of odor and mechanical stimuli to the sensory organs, as well as mass flow processes including heat loss and gas exchange in flying animals. With recent advances in near-field aerodynamics of insect and bird flight, it is now possible to determine how wing kinematics affects induced flow over their body. In this paper, I develop a theoretical model based in rotor theory to estimate the mean induced flow over the body of flapping insects. This model is able to capture some key characteristics of mean induced flow over the body of a flying insect. Specifically, it predicts that induced flow is directly proportional to wing beat frequency and stroke amplitude and is also affected by a wing shape dependent parameter. The derivation of induced flow includes the determination of spanwise variation of circulation on flapping wings. These predictions are tested against the available data on the spanwise distribution of aerodynamic circulation along finite Drosophila melanogaster wings and mean flows over the body of Manduca sexta. To explicitly account for tip losses in finite wings, a formula previously proposed by Prandtl for a finite blade propeller system is tentatively included. Thus, the model described in this paper allows us to estimate how far-field flows are influenced by near-field events in flapping flight.

Air Movements↗

Induced airflow in flying insects II. Measurement of induced flow.

The flapping wings of insects and birds induce a strong flow over their body during flight. Although this flow influences the sensory biology and physiology of a flying animal, there are very little data on the characteristics of this self-generated flow field or its biological consequences. A model proposed in the companion paper estimated the induced flow over flying insects. In this study, we used a pair of hot wire anemometers to measure this flow at two locations near the body of a tethered flapping hawk moth, Manduca sexta. The axial inflow anemometer measured the airflow prior to its entry into the stroke plane, whereas the radial outflow anemometer measured the airflow after it crossed the stroke plane. The high temporal resolution of the hot wire anemometers allowed us to measure not only the mean induced flow but also subtle higher frequency disturbances occurring at 1-4 times the wing beat frequency. These data provide evidence for the predictions of a mathematical model proposed in the companion paper. Specifically, the absolute value of the measured induced flow matches the estimate of the model. Also, as predicted by the model, the induced flow varies linearly with wing beat frequency. Our experiments also show that wing flexion contributes significantly to the observed higher frequency disturbances. Thus, the hot wire anemometry technique provides a useful means to quantify the aerodynamic signature of wing flexion. The phasic and tonic components of induced flow influence several physiological processes such as convective heat loss and gas exchange in endothermic insects, as well as alter the nature of mechanosensory and olfactory stimuli to the sensory organs of a flying insect.

Air Movements↗

Antibodies to mammalian and plant V-ATPases cross react with the V-ATPase of insect cation-transporting plasma membranes.

In immunobiochemical blots, polyclonal antibodies against subunits of plant and mammalian vacuolar-type ATPases (V-ATPases) cross-react strongly with corresponding subunits of larval Manduca sexta midgut plasma membrane V-ATPase. Thus, rabbit antiserum against Kalanchoe daigremontiana tonoplast V-ATPase holoenzyme cross-reacts with the 67, 56, 40, 28 and 20 kDa subunits of midgut V-ATPase separated by SDS-PAGE. Antisera against bovine chromaffin granule 72 and 39 kDa V-ATPase subunits cross-react with the corresponding 67 and 43 kDa subunits of midgut V-ATPase. Antisera against the 57 kDa subunit of both beet root and oat root V-ATPase cross-react strongly with the midgut 56 kDa V-ATPase subunit. In immunocytochemical light micrographs, antiserum against the beet root 57 kDa V-ATPase subunit labels the goblet cell apical membrane of both posterior and anterior midgut in freeze-substituted and fixed sections. The plant antiserum also labels the apical brush-border plasma membrane of Malpighian tubules. The ability of antibodies against plant V-ATPase to label these insect membranes suggests a high sequence homology between V-ATPases from plants and insects. Both of the antibody-labelled insect membranes transport K+ and both membranes possess F1-like particles, portasomes, on their cytoplasmic surfaces. This immunolabelling by xenic V-ATPase antisera of two insect cation-transporting membranes suggests that the portasomes on these membranes may be V-ATPase particles, similar to those reported on V-ATPase-containing vacuolar membranes from various sources.

Adenosine Triphosphatases↗

The novel aerodynamics of insect flight: applications to micro-air vehicles.

The wing motion in free flight has been described for insects ranging from 1 to 100 mm in wingspan. To support the body weight, the wings typically produce 2-3 times more lift than can be accounted for by conventional aerodynamics. Some insects use the fling mechanism: the wings are clapped together and then flung open before the start of the downstroke, creating a lift-enhancing vortex around each wing. Most insects, however, rely on a leading-edge vortex (LEV) created by dynamic stall during flapping; a strong spanwise flow is also generated by the pressure gradients on the flapping wing, causing the LEV to spiral out to the wingtip. Technical applications of the fling are limited by the mechanical damage that accompanies repeated clapping of the wings, but the spiral LEV can be used to augment the lift production of propellers, rotors and micro-air vehicles (MAVs). Design characteristics of insect-based flying machines are presented, along with estimates of the mass supported, the mechanical power requirement and maximum flight speeds over a wide range of sizes and frequencies. To support a given mass, larger machines need less power, but smaller ones operating at higher frequencies will reach faster speeds.

Animals↗

Selective adaptation to noxious foods by a herbivorous insect.

When animals repeatedly sample a noxious food over a period of 1-4 days, they can markedly reduce their aversive behavioral response to the diet's unpleasant taste (e.g. 'bitterness') or toxic effects. This long-term adaptation process is selective, however, permitting insects to adapt physiologically to some but not all noxious foods. We hypothesized (i) that the selective nature of this adaptation process stems from the fact that some unpalatable foods are toxic while others are harmless and (ii) that insects have more difficulty adapting to foods that are both unpalatable and toxic. Our model system consisted of Manduca sexta caterpillars and two compounds that taste bitter to humans and elicit an aversive behavioral response in this insect (salicin and aristolochic acid). We found that 2 days of exposure to a salicin diet completely adapted the aversive response of the caterpillars to salicin, but that exposure to an aristolochic acid diet failed to adapt the aversive response to aristolochic acid. We determined that M. sexta could not adapt to the aristolochic acid diet because it lacked mechanisms for reducing the compound's toxicity. In contrast, the salicin diet did not produce any apparent toxic effects, and the caterpillars adapted to its aversive taste within 12 h of exposure. We also found that the salicin adaptation phenomenon (i) was mediated by the central gustatory system, (ii) generalized to salicin concentrations that were twice those in the adapting diet and (iii) offset spontaneously when the caterpillar was transferred to a salicin-free diet. We propose that toxicity is a more significant barrier to dietary adaptation than 'bitterness' in this insect.

Adaptation, Physiological↗

Jumping in a winged stick insect.

The Thailand winged stick insect (Sipyloidea sp.) flees rapidly from a disturbance by jumping forwards when stimulated on the abdomen and backwards when stimulated on the head. The mechanisms underlying these fast movements were analysed by measuring movements of the body and legs from images captured at 250 Hz. A forward jump of both adults and nymphs involves movements of the abdomen and the middle and hind pairs of legs. The abdomen is raised and swung forwards by flexion at the joint with the metathorax and at the joint between the meso- and metathorax. At the same time, the tibiae of the hind and middle legs are extended and their femora depressed. The femoro-tibial joints of the legs are not fully flexed before a jump, and no structures in these joints appear to store muscular energy. The whole jumping sequence takes approximately 100 ms and results in take-off angles of 10-35 degrees at velocities of 0.6-0.8 m s(-1) and with an acceleration of 10 m s(-2). The abdominal angular velocity was 2000 degrees s(-1) and the tip of the abdomen moved at linear velocities of some 1 m s(-1), while the maximum rate of tibial extension was 4000 degrees s(-1). Rapid backward movements result either in the collapse of the body onto the ground, with a displacement away from the stimulus of approximately half a body length, or in the propulsion of the insect off its perch. Neither movement involves curling of the abdomen. From a horizontal posture, the forward jumps result in a displacement of a few body lengths. More lift can be generated in adults by elevating the hind wings as the abdomen is swung forwards and depressing them as the legs lose contact with the ground. In this way, jumps can lead directly to flapping flight. Take-off into flight can, however, be achieved without the abdominal movements seen during jumping. From a vertical posture, a forward jump propels the insect upwards and backwards before it falls to the ground horizontally displaced from its perch. Backward movements result in the insect falling with little horizontal displacement from its perch.

Animals↗

Water balance across the cuticle of a soil insect.

1. The water regime in soil commonly approaches equilibrium of water potential with the insects living there. 2. Even under these conditions, non-equilibrium processes have a significant effect on water movement through the cuticle of soil insects. 3. Measurements of water potential on either side of the cuticle of Costelytra zealandica larvae showed that equilibrium is not reached while the insect is alive. There is an active outward flow of water by thermoosmosis associated with the flow of heat from the insect.

Animals↗

Proteolytic activity and recombinant protein production in virus-infected Sf-9 insect cell cultures supplemented with carboxyl and cysteine protease inhibitors.

In insect cell-baculovirus expression systems for recombinant protein production, it is sometimes necessary to supplement cultures with protease inhibitors to protect recombinant proteins against proteolysis. To date, however, there is no information available concerning protease activities in inhibitor-supplemented cultures. The aim of the present study was to investigate intracellular and extracellular protease activities in cultures of virus-infected Sf-9 insect cells which were supplemented with inhibitors against carboxyl and cysteine proteases produced during culture. Prior to the supplementation culture, the cell toxicity of several protease inhibitors was determined. As a result, pepstatin A (carboxyl protease inhibitor) and E64, cystatin, leupeptin, and antipain (cysteine protease inhibitors) tested in this study showed no apparent negative effects on the growth and viability of noninfected Sf-9 insect cells at low concentrations. In addition, E64 and pepstatin A could rapidly permeate virus-infected Sf-9 cells and inhibit the respective intracellular protease activities. A virus-infected culture with a multiplicity of infection of 1 was carried out with E64 and pepstatin A which were added to the culture medium at 2 d post-infection. As a result of inhibitor supplementation, the cellular activity for recombinant protein biosynthesis was reduced by 5-30%. However, a significant reduction in carboxyl and cysteine protease activities was observed not only in the medium but also intracellularly. This is the first study that directly demonstrates a reduction in extracellular and intracellular protease activities in protease inhibitor-supplemented cultures of virus-infected insect cells.

Journal Article↗

Why does insect antifreeze protein from Tenebrio molitor produce pyramidal ice crystallites?

The antifreeze protein (AFP) reduces the growth rates of the ice crystal facets. In that process the ice morphology undergoes a modification. An AFP-induced surface pinning mechanism, through matching of periodic bond chains in two dimensions, enables two-dimensional regular ice-binding surfaces (IBSs) of the insect AFPs to engage a certain class of ice surfaces, called primary surfaces. They are kinetically stable surfaces with unambiguous and predetermined orientations. In this work, the orientations and molecular compositions of the primary ice surfaces that undergo growth rate reduction by the insect AFPs are obtained from first principles. Besides the basal face and primary prism, the ice surfaces engaged by insect AFPs include the specific ice pyramids produced by the insect AFP Tenebrio molitor (TmAFP). TmAFP-induced pyramids differ fundamentally from the ice pyramids produced by fish AFPs and antifreeze protein glycoproteins (AFPGs) as regards the ice surface configurations and the mode of interaction with the protein IBS. The molecular compositions of the TmAFP-induced pyramids are strongly bonded in two dimensions and have the constant face indices (101). In contrast, the molecular composition of the ice pyramids produced by fish AFPs and AFPGs are strongly bonded in only one direction and have variable face indices (h 0 l), none of which equal (101). The thus far puzzling behavior of the TmAFP in producing pyramidal crystallites is fully explained in agreement with experiment.

Animals↗

Plasma membranes from insect midgut cells.

Plasma membranes from insect midgut cells are separated into apical and basolateral domains. The apical domain is usually modified into microvilli with a molecular structure similar to other animals. Nevertheless, the microvillar structure should differ in some insects to permit the traffic inside them of secretory vesicles that may budd laterally or pinch-off from the tips of microvilli. Other microvillar modifications are associated with proton-pumping or with the interplay with an ensheathing lipid membrane (the perimicrovilllar membrane) observed in the midgut cells of hemipterans (aphids and bugs). The perimicrovillar membranes are thought to be involved in amino acid absorption from diluted diets. The microvillar and perimicrovillar membranes have densities (and protein content) that depend on the insect taxon. The role played by the microvillar and perimicrovillar proteins in insect midgut physiology is reviewed here trying to provide a coherent picture of data and highlighting further research areas.

Animals↗