Wing Growth Research Articles

The NDNF-like factor Nord is a Hedgehog-induced extracellular BMP modulator that regulates Drosophila wing patterning and growth.

Hedgehog (Hh) and Bone Morphogenetic Proteins (BMPs) pattern the developing Drosophila wing by functioning as short- and long-range morphogens, respectively. Here, we show that a previously unknown Hh-dependent mechanism fine-tunes the activity of BMPs. Through genome-wide expression profiling of the Drosophila wing imaginal discs, we identify nord as a novel target gene of the Hh signaling pathway. Nord is related to the vertebrate Neuron-Derived Neurotrophic Factor (NDNF) involved in congenital hypogonadotropic hypogonadism and several types of cancer. Loss- and gain-of-function analyses implicate Nord in the regulation of wing growth and proper crossvein patterning. At the molecular level, we present biochemical evidence that Nord is a secreted BMP-binding protein and localizes to the extracellular matrix. Nord binds to Decapentaplegic (Dpp) or the heterodimer Dpp-Glass-bottom boat (Gbb) to modulate their release and activity. Furthermore, we demonstrate that Nord is a dosage-dependent BMP modulator, where low levels of Nord promote and high levels inhibit BMP signaling. Taken together, we propose that Hh-induced Nord expression fine-tunes both the range and strength of BMP signaling in the developing Drosophila wing.

The Hox gene Antennapedia is essential for wing development in insects.

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 20-hydroxyecdysone. Additional targets of Antp are wing cuticular protein genes CPG24, CPH28 and CPG9, which are essential for wing development. We propose, therefore, that insect wing development occurs in an Antp-dependent manner. This article has an associated 'The people behind the papers' interview.

Components of environment affecting the reproduction of powder beetle Sitophilus zeamais (motsch.)

Several factors that significantly affect the reproduction of powder post beetles were food, moisture content, temperature, humidity, light, air/aeration, and the presence of natural enemies. This paper reviews the environment of the components that affect the reproduction of the powder post beetles Sitophilus zeamasi Motsch. Carbohydrates (sucrose and fructose) are the most significant energy source for the reproductive system and life span. In contrast, proteins consisting of amino acids such as arginine, lysine, leucine, isoleucine, tryptophan, histidine, phenylalanine, methionine, valine, and threonine, as well as components of fat, fatty acids, and sterols, are needed by insects for their energy supply and wing growth. The development of the powder post beetle population is rapid if the moisture content of the stored material is more than 15%. Temperature is also very influential on the reproduction of powder post beetles. The optimum humidity for this pest is around 75%. The next component is light, whereas light (dark and bright conditions) affects insect behavior in choosing food and reproducing (copulation and laying eggs). Another component is oxygen, whereby oxygen levels in the air must be more than 2%. If the CO2 level in the storage room is above 40%, then the pest in all stages will perish.

PVHL-mediated SMAD3 degradation suppresses TGF-β signaling.

Transforming growth factor β (TGF-β) signaling plays a fundamental role in metazoan development and tissue homeostasis. However, the molecular mechanisms concerning the ubiquitin-related dynamic regulation of TGF-β signaling are not thoroughly understood. Using a combination of proteomics and an siRNA screen, we identify pVHL as an E3 ligase for SMAD3 ubiquitination. We show that pVHL directly interacts with conserved lysine and proline residues in the MH2 domain of SMAD3, triggering degradation. As a result, the level of pVHL expression negatively correlates with the expression and activity of SMAD3 in cells, Drosophila wing, and patient tissues. In Drosophila, loss of pVHL leads to the up-regulation of TGF-β targets visible in a downward wing blade phenotype, which is rescued by inhibition of SMAD activity. Drosophila pVHL expression exhibited ectopic veinlets and reduced wing growth in a similar manner as upon loss of TGF-β/SMAD signaling. Thus, our study demonstrates a conserved role of pVHL in the regulation of TGF-β/SMAD3 signaling in human cells and Drosophila wing development.

Asymmetric requirement of Dpp/BMP morphogen dispersal in the Drosophila wing disc

How morphogen gradients control patterning and growth in developing tissues remains largely unknown due to lack of tools manipulating morphogen gradients. Here, we generate two membrane-tethered protein binders that manipulate different aspects of Decapentaplegic (Dpp), a morphogen required for overall patterning and growth of the Drosophila wing. One is “HA trap” based on a single-chain variable fragment (scFv) against the HA tag that traps HA-Dpp to mainly block its dispersal, the other is “Dpp trap” based on a Designed Ankyrin Repeat Protein (DARPin) against Dpp that traps Dpp to block both its dispersal and signaling. Using these tools, we found that, while posterior patterning and growth require Dpp dispersal, anterior patterning and growth largely proceed without Dpp dispersal. We show that dpp transcriptional refinement from an initially uniform to a localized expression and persistent signaling in transient dpp source cells render the anterior compartment robust against the absence of Dpp dispersal. Furthermore, despite a critical requirement of dpp for the overall wing growth, neither Dpp dispersal nor direct signaling is critical for lateral wing growth after wing pouch specification. These results challenge the long-standing dogma that Dpp dispersal is strictly required to control and coordinate overall wing patterning and growth.

Expression of Human Mutant Preproinsulins Induced Unfolded Protein Response, Gadd45 Expression, JAK-STAT Activation, and Growth Inhibition in Drosophila.

Mutations in the insulin gene (INS) are frequently associated with human permanent neonatal diabetes mellitus. However, the mechanisms underlying the onset of this genetic disease is not sufficiently decoded. We induced expression of two types of human mutant INSs in Drosophila using its ectopic expression system and investigated the resultant responses in development. Expression of the wild-type preproinsulin in the insulin-producing cells (IPCs) throughout the larval stage led to a stimulation of the overall and wing growth. However, ectopic expression of human mutant preproinsulins, hINSC96Y and hINSLB15YB16delinsH, neither of which secreted from the β-cells, could not stimulate the Drosophila growth. Furthermore, neither of the mutant polypeptides induced caspase activation leading to apoptosis. Instead, they induced expression of several markers indicating the activation of unfolded protein response, such as ER stress-dependent Xbp1 mRNA splicing and ER chaperone induction. We newly found that the mutant polypeptides induced the expression of Growth arrest and DNA-damage-inducible 45 (Gadd45) in imaginal disc cells. ER stress induced by hINSC96Y also activated the JAK-STAT signaling, involved in inflammatory responses. Collectively, we speculate that the diabetes-like growth defects appeared as a consequence of the human mutant preproinsulin expression was involved in dysfunction of the IPCs, rather than apoptosis.

Effect of Moisture Stress on the Development of False Codling Moth Thaumatotibia leucotreta (Lepidoptera: Tortricidae) on Two Tomato Varieties.

False codling moth Thaumatotibia leucotreta (Meyrick) is a pest native to sub-Saharan Africa infesting over 100 plant species including tomato. Its survival and performance are influenced by changes in precipitation and stress-related biochemical changes in the host plant. Water availability for agricultural production continues to decline due to climate change affecting drought-sensitive crops like tomatoes. Little is known on host plant-insect interactions of T. leucotreta and tomato as influenced by moisture stress. Hence, our study tested the impact of different water holding capacities (WHC) (40%, 50%, 60%, 70%, and 90%) of plant growing media on the growth of Anna F1 and Yaye tomato varieties and the infestation rate and development of T. leucotreta on the two varieties. WHC significantly influenced the growth of Anna F1 and Yaye tomato varieties. WHC significantly affected stem girth of Anna F1 variety and leaf length, leaf width, stem girth, and plant height of the Yaye variety. For Yaye variety, T. leucotreta laid a significantly high number of eggs when grown at 70% WHC and had the highest pupation when grown at 60% WHC. The development of T. leucotreta as observed on wing growth was highest at 40% and 50% WHC for both Anna F1 variety and Yaye variety. Our study shows that the infestation of tomato by T. leucotreta is likely to be high when grown in water-scarce media. The results are useful for predicting possible future T. leucotreta trends with increasing water scarcity due to climate change and in designing pest management programmes.

Yorkie ensures robust tissue growth in Drosophila ribosomal protein mutants.

Heterozygosity of ribosomal protein genes causes a variety of developmental abnormalities in humans, which are collectively known as ribosomopathies, yet the underlying mechanisms remain elusive. Here, we analyzed Drosophila Minute (M)/+ mutants, a group of mutants heterozygous for ribosomal protein genes that exhibit a characteristic thin-bristle phenotype. We found that, although M/+ flies develop essentially normal wings, simultaneous deletion of one copy of the Hippo pathway effector yki resulted in severe wing growth defects. These defects were caused by JNK-mediated cell death in the wing pouch via Eiger/TNF signaling. The JNK activation in M/+, yki/+ wing discs required the caspase Dronc, which is normally blocked by DIAP1. Notably, heterozygosity of yki reduced DIAP1 expression in the wing pouch, leading to elevation of Dronc activity. Dronc and JNK formed a positive-feedback loop that amplifies Dronc activation, leading to apoptosis. Our observations suggest a mechanism of robust tissue growth whereby tissues with reduced ribosomal protein prevent ectopic apoptosis via Yki activity.

A unified mechanism for the control of Drosophila wing growth by the morphogens Decapentaplegic and Wingless.

Development of the Drosophila wing-a paradigm of organ development-is governed by 2 morphogens, Decapentaplegic (Dpp, a BMP) and Wingless (Wg, a Wnt). Both proteins are produced by defined subpopulations of cells and spread outwards, forming gradients that control gene expression and cell pattern as a function of concentration. They also control growth, but how is unknown. Most studies have focused on Dpp and yielded disparate models in which cells throughout the wing grow at similar rates in response to the grade or temporal change in Dpp concentration or to the different amounts of Dpp "equalized" by molecular or mechanical feedbacks. In contrast, a model for Wg posits that growth is governed by a progressive expansion in morphogen range, via a mechanism in which a minimum threshold of Wg sustains the growth of cells within the wing and recruits surrounding "pre-wing" cells to grow and enter the wing. This mechanism depends on the capacity of Wg to fuel the autoregulation of vestigial (vg)-the selector gene that specifies the wing state-both to sustain vg expression in wing cells and by a feed-forward (FF) circuit of Fat (Ft)/Dachsous (Ds) protocadherin signaling to induce vg expression in neighboring pre-wing cells. Here, we have subjected Dpp to the same experimental tests used to elucidate the Wg model and find that it behaves indistinguishably. Hence, we posit that both morphogens act together, via a common mechanism, to control wing growth as a function of morphogen range.

Discovery of wing imaginal discs in the penultimate instar of the lacewing Mallada desjardinsi (Insecta: Neuroptera: Chrysopidae) with histological notes on postembryonic imaginal disc development.

Holometabolous insects are alternatively named "Endopterygota" because, in the larvae of many taxa, the wing primordia in the lateral regions of the meso- and metathoracic segments form more or less invaginated structures called wing imaginal discs. Holometabolous insects exhibit differential developmental timing of the wing during ontogeny. The condition in which wing growth is deferred until the end of larval life has been considered ancestral, whereas early disc formation has been recognized as the derived condition. Even though wing disc development in holometabolous insects has been studied extensively in select groups, many questions remain about the development of the wing imaginal disc in the orders Raphidioptera, Megaloptera, Neuroptera, and Mecoptera. To clarify whether the wing imaginal disc of Neuroptera is typical of the derived condition, we examined the ontogeny of the wing imaginal discs in the lacewing Mallada desjardinsi histologically. Using both light microscopy and transmission electron microscopy, we were able to recognize wing imaginal discs in the penultimate larval instar (prefinal larval instar) of this species. To date, neuropteran insects have been characterized as having late-forming wing imaginal discs. However, our findings show that the developmental pattern of the wing imaginal discs within the Neuroptera represents a more derived pattern of development in the Holometabola than was assumed previously.

The Ras/MAPK pathway is required for regenerative growth of wing discs in the black cutworm Agrotis ypsilon

Regeneration is a common phenomenon in various organisms by which tissues restore the damaged or naturally detached parts. In insects, appendage regeneration takes place during the embryonic, larval and pupal stages for individual survival. The wing disc of black cutworm Agrotis ypsilon has the capacity of regeneration after ablation, but understanding of molecular mechanisms in wing disc regeneration is still limited. After ablation of partial or whole wing discs before the fifth instar larval stage, the adult wings appeared to be normal. In the last two larval stages, ablation of the left wing disc led to smaller corresponding adult wing. Cell proliferation was reduced in the ablated wing disc but was gradually recovered two days post ablation. Transcriptome analysis found that genes in the mitogen-activated protein kinase (MAPK) pathway were upregulated. Repression of gene expression in this pathway, including Ras oncogene at 64B (Ras64B), Downstream of raf1 (Dsor1), and cAMP-dependent protein kinase catalytic subunit 3 (Pka-C3) by RNA interference after ablation, led to diminishment of both adult wings, suggesting that the MAPK signaling is essential for wing growth. Additionally, cell proliferation was still decelerated by injecting Ras64B, Dsor, or Pka-C3 dsRNA two days after ablation, indicating that the MAPK signaling-regulated cell proliferation is essential for growth. These results provide molecular clues to the regulation of cell proliferation during regeneration in lepidopteran insects.

Effects of human disturbance on postnatal growth and baseline corticosterone in a long-lived bird.

Prolonged or repeated episodes of environmental stress could be especially detrimental for developing young, via impaired growth or development. Despite this, most studies investigating the effects of human recreational and tourism activities have focused on adults. An increasing demand for nature-based tourism in remote locations means that many seabirds, which have evolved largely in the absence of predators and humans, are being exposed to novel pressures. The slow-growing semi-precocial nestlings of the European storm petrel Hydrobates pelagicus experience higher mortality rates in nests exposed to human recreational disturbance. Here, we examine whether surviving nestlings reared in disturbed areas are also affected via changes in growth trajectories and baseline circulating glucocorticoids. Nestlings reared in high-disturbance areas displayed delayed mass growth, and we found weak evidence for slower rates of mass gain and tarsus growth, compared with nestlings reared in undisturbed areas. There were no differences in wing growth, consistent with prioritization of long wings, important for post-fledging survival. A tendency for a less marked age-related decline in corticosterone (CORT) in disturbed nestlings offers limited evidence that changes in growth trajectories were mediated by baseline CORT. However, disturbed nestlings could have experienced overall higher GC exposure if the acute GC response was elevated. 'Catch-up' growth enabled high-disturbance nestlings to overcome early constraints and achieve a similar, or even larger, asymptotic body size and mass as low-disturbance nestlings. While catch-up growth has been shown to carry costs for parents and offspring, the effects of disturbance were slight and considerably smaller than growth alterations driven by variation in environmental conditions between years. Nonetheless, effects of human recreational activities could be exacerbated under higher levels of human disturbance or in the presence of multiple pressures, as imposed by present rapid rates of environmental change.

Eclosion hormone functions in larva-pupa-adult ecdysis in Leptinotarsa decemlineata

In Drosophila melanogaster, ecdysis-triggering hormone (ETH) and eclosion hormone (EH) mutually enhance the release of each other through a positive feedback loop. Both are involved in the stimulation of ecdysis behavior series and molting-related physiological preparations. However, whether 20-hydroxyecdysone (20E) controls eh expression and whether EH plays a similar role in non-Drosophilid insects remain to be determined. In Leptinotarsa decemlineata, we found that Ldeh mRNA levels were high in the early and/or late stages and low in the intermediate period within each larval instar, indicating that the mRNA peaks positively correlate with 20E pulses. Consistent with the indication, we confirmed that 20E signaling stimulated the transcription of Ldeh. RNA interference (RNAi)-aided knockdown of Ldeh at the final instar stage slightly impaired pupation and significantly affected wing expansion. Around 20% of the Ldeh RNAi larvae remained as prepupae, completely wrapped in the old larval cuticles. These prepupae gradually darkened, dried and withered, and finally died. The remaining (about 80%) Ldeh hypomorphs became pupae and emerged as abnormal adults, bearing smaller and wrinkle elytra and hindwings. The adult tracheae in the Ldeh hypomorphs were full of liquid. In addition, the low pupation rate and high deformed adult proportion cannot be rescued by ingestion of either 20E or juvenile hormone. Thus, EH is critical for ecdysis and trachea clearance in L. decemlineata. The failure of trachea clearance may disenable air-swallowing during larva-pupa-adult transition and thus impacts wing growth and expansion.

Risk of predation on offspring reduces parental provisioning, but not flight performance or survival across early life stages

Abstract Developmental responses can help young animals reduce predation risk but can also yield costs to performance and survival in subsequent life stages with major implications for lifetime fitness. Compensatory mechanisms may evolve to offset such costs, but evidence from natural systems is largely lacking. In songbirds, increased nest predation risk should favour reduced provisioning, but also young that fledge (leave their nest) at an earlier age. Both responses can result in fledglings with shorter wings, reduced mobility and decreased survival. Young may compensate for shorter wings developmentally by reallocating resources towards feather development or behaviourally by adjusting flight kinematics or habitat use. However, underfed young may lack the capacity to express these phenotypes due to insufficient resources or an inability to adjust allocation of resources. Using predation risk experiments and 29 years of observational field data, we test whether increased nest predation risk reduces flight performance and survival during the fledgling stage and explore potential mechanisms that might underlie these effects. We show that young from high‐risk nests did not leave the nest earlier on average, but wing growth was slower likely due to observed reductions in parental feeding rates. Wings were shorter in high‐risk nests when fledglings left the nest early. Yet, fledglings from high‐risk nests showed improved flight performance for a given wing length such that flight performance at fledging did not differ between young from high‐risk and low‐risk nests. Young from high‐risk nests may have offset the costs of shorter wings on flight performance by accelerating the emergence of flight feathers from their sheaths to reduce wing porosity, though evidence for this mechanism was mixed. Fledglings from high‐risk nests also selected habitat with denser woody vegetation compared with young from low‐risk nests. Together, these developmental and behavioural responses seem to mitigate the expected effects of increased nest predation risk on fledgling survival. Ultimately, our results show that offspring predation risk can affect parental provisioning and offspring morphology without major implications for performance and survival in subsequent life stages. A free Plain Language Summary can be found within the Supporting Information of this article.

O-GlcNAcylation Dampens Dpp/BMP Signaling to Ensure Proper Drosophila Embryonic Development.

BMP (bone morphogenetic protein) signaling activity is precisely controlled by both pathway agonists andantagonists. Here, we identify a previously unrecognized BMP signaling antagonist. We demonstrate that the Drosophila BMP type I receptor Sax(Saxophone) functions as a Dpp (Decapentaplegic) receptor in Drosophila embryos, but that its activity is normally inhibited by the O-linked glycosyltransferase Sxc (Super sex combs). In wild-type embryos, Sax activity is inhibited, and the BMP type I receptor Tkv (Thickveins) is the sole conduit for Dpp. In contrast, in sxc mutants, the Dpp signal istransduced by both Tkv and Sax, and elevated Dpp signaling results in embryonic lethality. We also demonstrate that Sxc O-glycosylates Sax andobserve elevated Dpp signaling in response to maternal restriction of dietary sugar. These findings link fertility to nutritive environment and point to Sax signaling as the nutrient-sensitive branch of BMP signaling.

Weather effects on breeding parameters of two insectivorous passerines in a polluted area

Direct and indirect effects of environmental pollution affect negatively to birds' breeding performance in both urban and industrial environments, but much less is known on how pollution and natural stress factors work together. In our long-term study (1991–2018), we explored whether industrial pollution and associated habitat changes increase the sensitivity of breeding parameters (hatching and fledging success, nestling growth) to temperature and precipitation in two insectivorous bird species, the great tit (Parus major) and the pied flycatcher (Ficedula hypoleuca). We found that both species are rather weather sensitive in terms of their fledging success, but especially in the F. hypoleuca, the negative effect was strengthened in a polluted environment. For both study species, all the breeding parameters, except growth of F. hypoleuca nestlings, were inferior in the polluted area and negatively affected by cold weather. Independent of pollution, the duration of rainy spells during the nestling period had an overall negative effect on fledging success of F. hypoleuca, and this effect became stronger at cold temperatures. The length of rainy spells was, however, positively associated with nestling wing length in both species, possibly because of better availability of some important food resources for wing growth in more humid conditions. The weather-pollution interactions in our study populations were not overwhelmingly strong, but those found in F. hypoleuca show that such interactions exist, they are species-specific and in our study system most likely associated to pollution-related resource (e.g. food) limitation. Higher sensitivity of F. hypoleuca to low temperatures is likely related to its less well-insulated nests and higher dependence on aerial prey, the availability of which is especially reduced during cold and rainy spells. Our study indicates that anthropogenic stress, such as pollution, has synergistic effects with natural stress factors affecting passerine birds' breeding performance.

An inelastic-damage micromechanical model based on the wing- and secondary-cracking mechanisms under dynamic loading

For most rock materials, there exists a coupling between inelastic deformations caused by crack displacements on micro-crack faces and damage evolution due to nucleation and growth of wing- and secondary-cracks. While rock material is subjected to dynamic loading, the interaction between micro-cracks plays an important role in materials behavior. The self-consistent homogenization scheme is implemented in this paper to consider micro-cracks interaction and determine the equivalent mechanical properties of micro-cracked rock deteriorated by damage evolution. The aim of this article is to develop a self-consistent based micromechanical damage model by taking into account the wing- and secondary-cracking mechanisms accompanied by inelastic strains caused by crack displacements under dynamic compressive loading. While stress intensity factors in tensile and in-plane shear modes at flaw tips exceed the material fracture toughness in modes I and II, respectively, wing- and secondary-cracks are sprouted and damage evolution occurs. For closed cracks, an appropriate criterion for the secondary-crack initiation is proposed in this paper. Moreover, the evolution rule for the secondary-crack is determined based on the principles of dynamic fracture mechanics. The developed model algorithm is programmed in the commercial finite difference software environment for numerical simulation of rock material to investigate the relationship between the macroscopic mechanical behavior and the microstructure. The fracture toughness parameters of the rock samples are experimentally determined. The rock microstructure parameters (average initial length and density of flaws) are studied using scanning electron microscopy. In order to verify the developed model, a series of numerical simulations are carried out to numerically reproduce the Split-Hopkinson pressure bar test results. The numerical simulation results are in agreement with the experimental data. The simulation results demonstrate that the developed micromechanical model can adequately reproduce many features of the rock behavior such as softening in post-peak region, damage induced by wing- and secondary-cracks and irreversible deformations caused by crack displacements on micro-cracks. Furthermore, the softening behavior of rock material in the post-peak region is affected by considering inelasticity and the secondary-cracking mechanisms. Therefore, the rock sample simulation with the coupled inelastic-damage model can increase inelastic deformations in the post-peak region as a result of irreversible strains caused by crack displacements on micro-cracks. The simulation by considering the secondary-crack mechanism leads to an increase in the microcracking process and damage in rock material.

Cell Cycle Progression Determines Wing Morph in the Polyphenic Insect Nilaparvata lugens.

SummaryWing polyphenism is a phenomenon in which one genotype can produce two or more distinct wing phenotypes adapted to the particular environment. What remains unknown is how wing pad development is controlled downstream of endocrine signals such as insulin and JNK pathways. We show that genes important in cellular proliferation, cytokinesis, and cell cycle progression are necessary for growth and development of long wings. Wing pad cellular development of the long-winged morph was characterized by a highly structured epithelial layer with microvilli-like structures. Cells of adult short wing pads are largely in the G2/M phase of the cell cycle, whereas those of long wings are largely in G1. Our study is the first to report the comparative developmental and cellular morphology and structure of the wing morphs and to undertake a comprehensive evaluation of the cell cycle genes necessary for wing development of this unique, adaptive life history strategy.

Regulation of Anisotropic Tissue Growth by Two Orthogonal Signaling Centers.

The Drosophila wing has served as a paradigm to mechanistically characterize the role of morphogens in patterning and growth. Wingless (Wg) and Decapentaplegic (Dpp) are expressed in two orthogonal signaling centers, and their gradients organize patterning by regulating the expression of well-defined target genes. By contrast, graded activity of these morphogens is not an absolute requirement for wing growth. Despite their permissive role in regulating growth, here we show that Wg and Dpp are utilized in a non-interchangeable manner by the two existing orthogonal signaling centers to promote preferential growth along the two different axes of the developing wing. Our data indicate that these morphogens promote anisotropic growth by making use of distinct and non-interchangeable molecular mechanisms. Whereas Dpp drives growth along the anterior-posterior axis by maintaining Brinker levels below a growth-repressing threshold, Wg exerts its action along the proximal-distal axis through a double repression mechanism involving T cell factor (TCF).

Zika virus non-structural protein NS4A restricts eye growth in Drosophila through regulation of JAK/STAT signaling.

ABSTRACTTo gain a comprehensive view of the changes in host gene expression underlying Zika virus (ZIKV) pathogenesis, we performed whole-genome RNA sequencing (RNA-seq) of ZIKV-infected Drosophila adult flies. RNA-seq analysis revealed that ZIKV infection alters several and diverse biological processes, including stress, locomotion, lipid metabolism, imaginal disc morphogenesis and regulation of JAK/STAT signaling. To explore the interaction between ZIKV infection and JAK/STAT signaling regulation, we generated genetic constructs overexpressing ZIKV-specific non-structural proteins NS2A, NS2B, NS4A and NS4B. We found that ectopic expression of non-structural proteins in the developing Drosophila eye significantly restricts growth of the larval and adult eye and correlates with considerable repression of the in vivo JAK/STAT reporter, 10XStat92E-GFP. At the cellular level, eye growth defects are associated with reduced rate of proliferation without affecting the overall rate of apoptosis. In addition, ZIKV NS4A genetically interacts with the JAK/STAT signaling components; co-expression of NS4A along with the dominant-negative form of domeless or StatRNAi results in aggravated reduction in eye size, while co-expression of NS4A in HopTuml (also known as hopTum) mutant background partially rescues the hop-induced eye overgrowth phenotype. The function of ZIKV NS4A in regulating growth is maintained in the wing, where ZIKV NS4A overexpression in the pouch domain results in reduced growth linked with diminished expression of Notch targets, Wingless (Wg) and Cut, and the Notch reporter, NRE-GFP. Thus, our study provides evidence that ZIKV infection in Drosophila results in restricted growth of the developing eye and wing, wherein eye phenotype is induced through regulation of JAK/STAT signaling, whereas restricted wing growth is induced through regulation of Notch signaling. The interaction of ZIKV non-structural proteins with the conserved host signaling pathways further advance our understanding of ZIKV-induced pathogenesis.This article has an associated First Person interview with the first author of the paper.