Out from under the wing: reconceptualizing the insect wing gene regulatory network as a versatile, general module for body-wall lobes in arthropods

Body plan evolution often occurs through the differentiation of serially homologous body parts, particularly in the evolution of arthropod body plans. Recently, homeotic transformations resulting from experimental manipulation of gene expression, along with comparative data on the expression and function of genes in the wing regulatory network, have provided a new perspective on an old question in insect evolution: how did the insect wing evolve? We investigated the metamorphic roles of a suite of 10 wing- and body-wall-related genes in a hemimetabolous insect, Oncopeltus fasciatus . Our results indicate that genes involved in wing development in O. fasciatus play similar roles in the development of adult body-wall flattened cuticular evaginations. We found extensive functional similarity between the development of wings and other bilayered evaginations of the body wall. Overall, our results support the existence of a versatile development module for building bilayered cuticular epithelial structures that pre-dates the evolutionary origin of wings. We explore the consequences of reconceptualizing the canonical wing-patterning network as a bilayered body-wall patterning network, including consequences for long-standing debates about wing homology, the origin of wings and the origin of novel bilayered body-wall structures. We conclude by presenting three testable predictions that result from this reconceptualization.

Insect wing 3D printing

Insects have acquired various types of wings over their course of evolution and have become the most successful terrestrial animals. Consequently, the essence of their excellent environmental adaptability and locomotive ability should be clarified; a simple and versatile method to artificially reproduce the complex structure and various functions of these innumerable types of wings is necessary. This study presents a simple integral forming method for an insect-wing-type composite structure by 3D printing wing frames directly onto thin films. The artificial venation generation algorithm based on the centroidal Voronoi diagram, which can be observed in the wings of dragonflies, was used to design the complex mechanical properties of artificial wings. Furthermore, we implemented two representative functions found in actual insect wings: folding and coupling. The proposed crease pattern design software developed based on a beetle hindwing enables the 3D printing of foldable wings of any shape. In coupling-type wings, the forewing and hindwing are connected to form a single large wing during flight; these wings can be stored compactly by disconnecting and stacking them like cicada wings.

The Daphnia carapace and the origin of novel structures

SummaryUnderstanding how novel structures arise is a central question in evolution. The carapace of the waterflea Daphnia is a bivalved “cape” of exoskeleton that has been proposed to be one of many novel arthropod structures that arose through repeated co-option of genes that also pattern insect wings1–3. To determine whether the Daphnia carapace is a novel structure, we compare the expression of pannier, araucan, and vestigial between Daphnia, Parhyale, and Tribolium. Our results suggest that the Daphnia carapace did not arise by co-option, but instead derives from an ancestral exite (lateral lobe) that emerges from a proximal leg segment that was incorporated into the Daphnia body wall. The Daphnia carapace therefore appears to be homologous to the Parhyale tergal plate and the insect wing4. Remarkably, the vestigial-positive region that gives rise to the Daphnia carapace appears to be present in Parhyale5 and Tribolium as a small, inconspicuous protrusion. Similarly, the vestigial-positive developmental fields that form tergal plates in Parhyale appear to be present in Daphnia, even though Daphnia does not form tergal plates. Thus, rather than a novel structure resulting from gene co-option, the Daphnia carapace appears to have arisen from a shared, ancestral developmental field that persists in a cryptic state in other arthropod lineages. Cryptic persistence of unrecognized serially homologous developmental fields may thus be a general solution for the origin of novel structures. Our simple molecular triangulation strategy, which does not require functional studies, can illuminate the homologies of long-debated structures on the legs and body wall of arthropods.

Insect Wing Buckling Influences Stress and Stability During Collisions

Flapping insect wings frequently collide with vegetation and other obstacles during flight. Repeated collisions may irreversibly damage the insect wing, thereby compromising the insect’s ability to fly. Further, reaction torques caused by the collision may destabilize the insect and hinder its ability to maneuver. To mitigate the adverse effects of impact, some insect wings are equipped with a flexible joint called a “costal break.” The costal break buckles once it exceeds a critical angle, which is believed to improve flight stability and prevent irreversible wing damage. However, to our knowledge, there are no models to predict the dynamics of the costal break. Through this research, we develop a simple model of an insect wing with a costal break. The wing was modeled as two beams interconnected by a torsional spring, where the stiffness of the torsional spring instantaneously decreases once it has exceeded a critical angle. We conducted a series of static tests to approximate model parameters. Then, we used numerical simulation to estimate the peak stresses and reaction moments experienced by the wing during a collision. We found that costal break increased the wing’s natural frequency by about 50% compared to a homogeneous wing and thus reduced the stress associated with normal flapping. Buckling did not significantly affect peak stresses during collision. Joint buckling reduced the peak reaction moment by about 32%, suggesting that the costal break enhances flight stability.

Anthropogenic evolution in an insect wing polymorphism following widespread deforestation

Anthropogenic environmental change can underpin major shifts in natural selective regimes, and can thus alter the evolutionary trajectories of wild populations. However, little is known about the evolutionary impacts of deforestation—one of the most pervasive human-driven changes to terrestrial ecosystems globally. Absence of forest cover (i.e. exposure) has been suggested to play a role in selecting for insect flightlessness in montane ecosystems. Here, we capitalize on human-driven variation in alpine treeline elevation in New Zealand to test whether anthropogenic deforestation has caused shifts in the distributions of flight-capable and flightless phenotypes in a wing-polymorphic lineage of stoneflies from the Zelandoperla fenestrata species complex. Transect sampling revealed sharp transitions from flight-capable to flightless populations with increasing elevation. However, these phenotypic transitions were consistently delineated by the elevation of local treelines, rather than by absolute elevation, providing a novel example of human-driven evolution in response to recent deforestation. The inferred rapid shifts to flightlessness in newly deforested regions have implications for the evolution and conservation of invertebrate biodiversity.

Bmmp influences wing shape by regulating anterior-posterior and proximal-distal axis development

Insect wings are subject to strong selective pressure, resulting in the evolution of remarkably diverse wing shapes that largely determine flight capacity. However, the genetic basis and regulatory mechanisms underlying wing shape development are not well understood. The silkworm Bombyx mori micropterous ( mp ) mutant exhibits shortened wing length and enlarged vein spacings , albeit without changes in total wing area. Thus, the mp mutant comprises a valuable genetic resource for studying wing shape development. In this study, we used molecular mapping to identify the gene responsible for the mp phenotype and designated it Bmmp . Phenotype-causing mutations were identified as indels and single nucleotide polymorphisms in non-coding regions. These mutations resulted in decreased Bmmp mRNA levels and changes in transcript isoform composition. Bmmp null mutants were generated by CRISPR/Cas9 and exhibited significantly smaller wings. By examining the expression of genes critical to wing development in wildtype and Bmmp null mutants, we found that Bm mp exerts its function by coordinately modulating anterior-posterior and proximal-distal axis development. We also studied a Drosophila mp mutant and found that Bmmp is functionally conserved in Drosophila . The Drosophila mp mutant strain exhibits curly wings of reduced size and a complete loss of flight capacity. Our results increase our understanding of the mechanisms underpinning insect wing development and reveal potential targets for pest control.

The damping and structural properties of dragonfly and damselfly wings during dynamic movement

For flying insects, stability is essential to maintain the orientation and direction of motion in flight. Flight instability is caused by a variety of factors, such as intended abrupt flight manoeuvres and unwanted environmental disturbances. Although wings play a key role in insect flight stability, little is known about their oscillatory behaviour. Here we present the first systematic study of insect wing damping. We show that different wing regions have almost identical damping properties. The mean damping ratio of fresh wings is noticeably higher than that previously thought. Flight muscles and hemolymph have almost no ‘direct’ influence on the wing damping. In contrast, the involvement of the wing hinge can significantly increase damping. We also show that although desiccation reduces the wing damping ratio, rehydration leads to full recovery of damping properties after desiccation. Hence, we expect hemolymph to influence the wing damping indirectly, by continuously hydrating the wing system.

The Hox gene Antennapedia regulates wing development through 20-hydroxyecdysone in insect

A long-standing view in the field of evo-devo is that insect forewings develop without any Hox gene input. The Hox gene Antennapedia (Antp), despite being expressed in the thoracic segments of insects, has no effect on wing development. This view has been obtained from studies in two main model species, Drosophila and Tribolium. Here, we show that partial loss of function of Antp resulted in reduced and malformed adult wings in Bombyx, Drosophila, and Tribolium. Antp mediates wing growth in Bombyx by directly regulating the ecdysteriod biosynthesis enzyme gene (shade) in the wing tissue, which leads to local production of the growth hormone 20E. In turn, 20E signaling also up-regulates Antp. Additional targets of Antp are wing cuticular protein genes CPG24, CPH28, and CPG9, essential for wing development. We propose, thus, that insect wing development occurs in an Antp-dependent manner.