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A Vestigial:Scalloped TEA domain chimera rescues the wing phenotype of a scalloped mutation in Drosophila melanogaster.

The development of the Drosophila wing requires both scalloped and vestigial functions. Using a fusion between full-length Vestigial and the Scalloped TEA domain, the fusion protein can rescue scalloped wing mutations because within wing development, Scalloped and Vestigial cooperatively act as a transcription complex. Scalloped provides the necessary DNA binding function via the TEA domain and Vestigial promotes the activation of target genes. We also demonstrate that the putative nuclear localization signal contained in the TEA domain of Scalloped is likely responsible for the nuclear localization of Vestigial. The fusion protein is also capable of activating a known target gene of the native complex and thus represents a tool that will be helpful in rapidly identifying target genes of the Sd/Vg complex that are involved in wing differentiation. The functionality of the fusion suggests that only the TEA domain of Scalloped is critical for wing development and the rest of the protein (about 70%) is dispensable. This result is novel and should stimulate further studies of sd in other tissues in view of the fact that scalloped is a vital gene in Drosophila.

Amino Acid Sequence↗

Targeted expression of the signaling molecule decapentaplegic induces pattern duplications and growth alterations in Drosophila wings.

In the wing imaginal disc, the decapentaplegic (dpp) gene is expressed in a stripe of anterior cells near the anterior-posterior compartment boundary, and it is required solely in these cells for the entire disc to develop. In some viable segment polarity mutants, alterations in dpp expression have been demonstrated that correlate with changes in wing morphology. To test the hypothesis that the abnormal patterns of dpp expression are responsible directly for the mutant phenotypes, we have expressed dpp in ectopic places in wing imaginal discs, and we have found that dpp is able to cause overgrowth and pattern duplications in both anterior and posterior compartments of the wing disc. The alterations of the anterior compartment are strikingly similar to those observed in some viable segment polarity mutants. Thus, ectopic dpp alone can account for the phenotype of these mutants. We also show that ectopic expression of the segment polarity gene hedgehog (hh) gives similar morphological changes and activates dpp expression in the anterior compartment. This strongly suggests that the organizating activity of hh is mediated by dpp. We propose that the expression of dpp near the anterior-posterior compartment boundary is directed by the interaction between patched and hh, and that dpp itself could act as a general organizer of the patterning in the wing imaginal disc.

Animals↗

Elements of butterfly wing patterns.

The color patterns on the wings of butterflies are unique among animal color patterns in that the elements that make up the overall pattern are individuated. Unlike the spots and stripes of vertebrate color patterns, the elements of butterfly wing patterns have identities that can be traced from species to species, and typically across genera and families. Because of this identity it is possible to recognize homologies among pattern elements and to study their evolution and diversification. Individuated pattern elements evolved from non-individuated precursors by compartmentalization of the wing into areas that became developmentally autonomous with respect to color pattern formation. Developmental compartmentalization led to the evolution of serially repeated elements and the emergence of serial homology. In these compartments, serial homologues were able to acquire site-specific developmental regulation and this, in turn, allowed them to diverge morphologically. Compartmentalization of the wing also reduced the developmental correlation among pattern elements. The release from this developmental constraint, we believe, enabled the great evolutionary radiation of butterfly wing patterns. During pattern evolution, the same set of individual pattern elements is arranged in novel ways to produce species-specific patterns, including such adaptations as mimicry and camouflage.

Adaptation, Physiological↗

Pattern formation in the imaginal wing disc of Drosophila melanogaster: fate map, regeneration and duplication.

Fragments of the wing disc of Drosophila (fig. 2) were either injected into mature third instar larvae for immediate metamorphosis, or cultured in adult abdomens for seven days before being transferred to larvae for metamorphosis. The structures differentiated during metamorphosis were then analysed. The results of the first series of experiments were used to construct an accurate fate map of the disc, and those of the second series were used to determine the regenerative properties of the disc. The fate map (fig. 7) shows presumptive proximal parts (notum, pleura, and dorsal and ventral hinge) at the two ends of the disc, with presumptive distal wing parts in between. During metamorphosis the disc epithelium folds upon itself along the presumptive wing border, bringing dorsal and ventral wing and hinge surfaces into apposition. The wing surfaces occupy a much smaller relative area, and the hinge parts a much larger relative area, in the fate map than in the adult structure. The cultured fragments, in general, behaved in accordance with the rule that when two cut surfaces are created by cutting across the disc, regeneration occurs from one of the cut surfaces and duplication occurs from the other (fig. 14). It was possible to define a level in the longitudinal axis of the disc from which regeneration proceeds outwards. Cut surfaces facing away from this level show regeneration, while cut surfaces facing this level undergo duplication. Similar behavior was found for the transverse axis, and for two diagonal series of cuts. Some fragments with two cut edges could regenerate from one edge while duplicating from the other, whereas others could regenerate from two cut edges simultaneously. However, fragments with four cut edges showed incomplete regeneration, and a high tendency to duplicate even though regeneration in all directions might have been expected on the basis of the other experiments.

Animals↗

Case of unilateral wing formation in the female of the glowworm Lampyris noctiluca.

On July 27, 1999, the first author found a unilaterally winged adult glowworm in a park in the city of Mainz. Except for the wings on the left side, the specimen exhibited female characteristics that extended to external sexual appendages, the lantern and the gonads. The internal organization showed some remarkable differences between right (wingless) and left (winged) side. The right ovary contained three times more mature eggs than the left side and the volume of the corpus allatum of this side was about one-third larger than that of the left side. This suggests that aptery and egg maturation are affected by corpus allatum activity, i.e., juvenile hormone production. The findings do not support the hypothesis of Naisse ([1966] Arch Biol Liège 77:139-201) that wing formation, as a secondary male characteristic, is controlled by an androgenic hormone from the testes in the glowworm. Thus, the observations on this exceptional specimen have implications for the current hypotheses concerning the control of sexual wing dimorphism in Lampyris noctiluca.

Animals↗

PS2 integrin requirements in Drosophila embryo and wing morphogenesis.

The Drosophila inflated (if) gene encodes the alpha PS2 subunit of the PS integrins. We describe the generation of new if mutations, their lethal embryonic phenotype, and experiments that examine the spatial and temporal requirements for integrins in adult wing morphogenesis. Embryos hemizygous for either new allele, ifA7 or ifB2, make reduced amounts of alpha PS2. In a variety of genetic tests, these alleles behave similarly to ifk27e, which makes no detectable alpha PS2, and all three alleles display the same embryonic phenotype. We therefore conclude that all of the lethal alleles retain little or no wild-type alpha PS2 function. As seen for strong mutations at the myospheroid (mys) locus, which encodes the beta PS integrin subunit, if mutants show extreme defects in somatic muscle attachments and in midgut morphogenesis. Unlike mys, however, there is no dorsal herniation of the if mutant embryos. With respect to wing morphogenesis, clonal analysis experiments demonstrate that if+ function is required only in cells of the ventral wing surface. We have rescued the wing blister phenotype of double mutants for the hypomorphic mysnj42 and if3 alleles using a heat shock-inducible mys+ transgene. By varying times of transgene induction, we find that integrin function is required from very early in metamorphosis until at least the last 24-48 hr of wing development.

Alleles↗

Wingless modulates the effects of dominant negative notch molecules in the developing wing of Drosophila.

The development and patterning of the wing in Drosophila relies on a sequence of cell interactions molecularly driven by a number of ligands and receptors. Genetic analysis indicates that a receptor encoded by the Notch gene and a signal encoded by the wingless gene play a number of interdependent roles in this process and display very strong functional interactions. At certain times and places, during wing development, the expression of wingless requires Notch activity and that of its ligands Delta and Serrate. This has led to the proposal that all the interactions between Notch and wingless can be understood in terms of this regulatory relationship. Here we have tested this proposal by analysing interactions between Delta- and Serrate-activated Notch signalling and Wingless signalling during wing development and patterning. We find that the cell death caused by expressing dominant negative Notch molecules during wing development cannot be rescued by coexpressing Nintra. This suggests that the dominant negative Notch molecules cannot only disrupt Delta and Serrate signalling but can also disrupt signalling through another pathway. One possibility is the Wingless signalling pathway as the cell death caused by expressing dominant negative Notch molecules can be rescued by activating Wingless signalling. Furthermore, we observe that the outcome of the interactions between Notch and Wingless signalling differs when we activate Wingless signalling by expressing either Wingless itself or an activated form of the Armadillo. For example, the effect of expressing the activated form of Armadillo with a dominant negative Notch on the patterning of sense organ precursors in the wing resembles the effects of expressing Wingless alone. This result suggests that signalling activated by Wingless leads to two effects, a reduction of Notch signalling and an activation of Armadillo.

Animals↗

The hiiragi gene encodes a poly(A) polymerase, which controls the formation of the wing margin in Drosophila melanogaster.

The hiiragi (hrg) gene plays a key role in the development of the wing margin in Drosophila melanogaster. A mutation in the hrg gene resulted in a decrease in the level of the hrg transcript and was associated with a notched wing phenotype. We report here that the hrg gene encodes a poly(A) polymerase (PAP). The bovine cDNA for PAP type II reversed the phenotype due to mutation of the hrg gene, suggesting that hrg might encode a functional homolog of PAP. A mutation that reduced the enzymatic activity of Hrg failed to reverse the phenotype of hrg mutants, suggesting that the enzymatic activity of Hrg was required to rescue the wing phenotype. The levels of expression of wingless and cut at the presumptive wing margins were reduced in the late third-instar larvae of hrg mutants. These results suggest that the product of hrg is required for the normal expression of a series of genes in this region. Our results provide the first evidence that a PAP in Drosophila plays a key role in the early development of the wing margin, acting to regulate the specific expression of a series of genes via, perhaps, control of the processing of the 3' ends of transcripts.

Amino Acid Sequence↗

A scanning electron microscopic study of the normal development of the chick wing from stages 19 to 36. A supplement to the Hamburger and Hamilton staging system.

The purpose of a staging system is to stage embryos by morphology rather than chronology. This is particularly useful when embryos do not develop exactly synchronously, as in the case of the chick. At present the Hamburger and Hamilton (1951) series is universally used to stage chick embryos. The aim of the present study was to provide a series of morphological descriptions of the normal stages of development of the chick wing bud from stages 19 to 36, and to correct some errors of the original system which may be overlooked by those new to the chick wing bud as an experimental model, and who rely primarily on the Hamburger and Hamilton stage series. In addition, Summerbell's (1976) observations on the appearance of the cartilaginous elements made from alcian green-stained whole mounts have been correlated with the external appearance of the wing bud to provide a more complete understanding of the skeletal development that influences, and to some degree accounts for, the changes in external morphology. Scanning electron microscopy (SEM) has been used to obtain images of much greater resolution and detail than those available from Hamburger and Hamilton, whilst using comparable magnifications to those attainable using conventional dissecting microscopes. The number of somites across which the proximal part of the wing bud extends has been provided as a measure of the limb width at early stages (19-24). At certain stages there were clear differences between the characteristic wing bud features described by Hamburger and Hamilton and those observed in the present study.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The control of wing kinematics by two steering muscles of the blowfly (Calliphora vicina).

We used a combination of high speed video and electrophysiological recordings to investigate the relationship between wing kinematics and the firing patterns of the first (b1) and second (b2) basalar muscles of tethered flying blowflies (Calliphora vicina). The b1 typically fires once during every wing stroke near the time of the dorsal stroke reversal. The b2 fires either intermittenly or in bursts that may be elicited by a visual turning stimulus. Sustained activation of the b1 at rates near wing beat frequency appears necessary for tonic maintenance of stroke amplitude. In addition, advances in the phase of b1 activation were correlated with both increased wing protraction during the down-stroke and increased stroke amplitude. Similar kinematic alterations was correlated with b2 spikes, and consequently, both muscles may function in the control of turns toward the contralateral side. The effects of the two muscles were evident within a single stroke period and decayed quickly. Kinematic changes correlated with b1 phase shifts were graded, suggesting a role in compensatory course stabilization. In contrast, b2 spikes were correlated with all-or-none changes in the wing stroke, a characteristic consistent with a role in mediating rapid turns towards or away from objects.

Animals↗

In vitro studies on the existence of endothelial precursor cells in the subectodermal avascular region of quail wing buds.

In vitro studies were performed to investigate the angiogenic capacity of different parts of the avian limb bud. Small pieces of wing mesenchyme of the vascularized core or of the avascular subectodermal region were obtained from quail embryos at stages 18 to 25, and were cultured. The identification of the avascular wing mesenchyme was made possible after injection of India ink via the vitelline vein or by bleeding control during in vivo dissection. Tissue cultures were treated with the QH-1 antibody or/and the endothelial cell marker DiI-Ac-LDL. Endothelial cells were found in cultures of the mesenchymal core and in those of the avascular subectodermal wing mesenchyme. Moreover, their appearance was independent of the stage of the donor embryo. Although there were no vessels, the subectodermal wing mesenchyme was able to produce endothelial cells that proliferated and differentiated under in vitro conditions. Thus, endothelial precursor cells probably existed within the avascular wing mesenchyme. These cells might be identical with the QH-1-positive isolated cells that have been described in immunohistochemical studies of this region; they may contribute to the growing capillary plexus of the limb bud.

Animals↗

Experimental analysis of the origin of the wing musculature in avian embryos.

Interspecific grafts of somites, as well as parts of the somatic plate mesoderm, have been made between quail and chick embryos (stages 12--14 H. H.) at the level of the prospective wing bud in order to examine the relationship between somites and wing bud myogenesis. The stability of the natural quail nuclear labelling makes it possible to follow the developmental fate of grafted mesodermal cells in the host embryo. Embryos examined after subsequent incubation periods of 3--7 days show the following distribution of somatic and somitic cells within the wing bud: as soon as the three zones of different cell density within the mesoderm can be distinguished, cells of somitic origin are limited to the prospective myogenic area which is made up of mixed population of somatic and somitic cells, whereas the prospective chondrogenic area as well as the subectodermal zone only consists of cells originated from the somatic plate mesoderm. After further incubation, single muscle blastema are present which were also seen to be a mixture of somatic and somitic cells. The cells of muscular bundles are of somitic origin, while the muscle connective tissue cells are derived from the somatic plate mesoderm. After grafting into the coelomic cavity or on the chorio-allantoic membrane, fragments of the somatic plate mesoderm previously isolated from quail embryos (stage 14 H.H.) at the level of the prospective wing bud exhibit well developed skeletal elements, but fail to differentiate any musculature. These experimental investigations support previous evidence for a somitic origin of wing bud myogenic cells. Histological and scanning electron microscopic studies of the brachial somites and the adjacent somatic plate mesoderm of chick embryo (stages 13--15 H.H.) reveal that migration of still undifferentiated somitic cells into the brachial somatic plate mesoderm begins to take place in embryos at stage 14.

Animals↗

On the migration of epidermal melanoblasts in the avian embryonic wing bud.

After heterotopic grafting of quail neural crest cells to the wing buds of embryos of an unpigmented chicken strain, epidermal melanocytes of donor origin are found almost exclusively distal from the graft in the host's epidermis. This directed cell migration ceases, if the apical ectodermal ridge (together with a small amount of subridge mesoderm) is removed from the operated wing buds or if impermeable materials are interposed between it and the rest of the wing bud. Under these conditions epidermal melanocytes are found not only distal from but also proximal to the grafts. From this it may be deduced that the apical ectodermal ridge directs the migration of epidermal melanoblasts in the avian embryonic wing bud, possibly by a chemotactic mechanism. The presence or absence of the apical ectodermal ridge had no observable effect on the migratory behaviour of other neural crest derived cell populations (Schwann cells and non-epidermal melanocytes) in the wing bud. This shows that the apical ectodermal ridge specifically influences epidermal melanocytes.

Animals↗

Genotoxic evaluation of ten carcinogens in the Drosophila melanogaster wing spot test.

To provide further background data on the wing spot somatic mutation and recombination assay, 10 selected carcinogens (acetamide, acrylamide, benzo(a)pyrene, cyclophosphamide, diethylstilbestrol, 4-nitroquinoline N-oxide, propyleneimine, safrole, thiourea, and o-toluidine) were tested in this assay. 72-h-old third-instar larvae, trans-heterozygous for 2 recessive wing cell markers: multiple wing hairs (mwh) and flare3 (flr3) were fed with 3 concentrations of each carcinogen during the rest of their development until pupation, and the genotoxic effects were measured as significant increases in the appearance of visible mutant hair clones on the adult wing blade. Our results show that 6 of the carcinogens tested produce significant increases in wing spot frequency, at least at one of the concentrations assayed. Benzo(a)pyrene, diethylstilbestrol, safrole and thiourea were the compounds that did not increase the incidence of mutant clones.

4-Nitroquinoline-1-oxide↗

The effect of female wings on male courtship behavior in the cricket Gryllus bimaculatus.

Sexual dimorphism exists in the shape and the structure of the forewings of the cricket (Gryllus bimaculatus). However, the functional significance of the wings in the G. bimaculatus female has been unclear. In common blue butterflies (Polyommatus icarus), wings in females have been suggested as being important for attracting males. To test whether female crickets need wings for conspecific males to recognize them and initiate mating behavior, we removed all wings from females and observed the behavior of males towards them. Most males (87.5%) showed mating behavior towards the wingless females: they produced courtship song and transferred spermatophores to the wingless females. Similarly, 88.5% of the males showed mating behavior towards intact females. When males were placed with both a wingless female and an intact female, no significant difference was detected in male mate choice. The findings demonstrate that the wing of the G. bimaculatus female is not necessary for female recognition by conspecific males and the initiation of male mating behavior, and that it is not important in male mate choice.

Animals↗

Acoustic startle/escape reactions in tethered flying locusts: motor patterns and wing kinematics underlying intentional steering.

We simultaneously recorded flight muscle activity and wing kinematics in tethered, flying locusts to determine the relationship between asymmetric depressor muscle activation and the kinematics of the stroke reversal at the onset of wing depression during attempted intentional steering manoeuvres. High-frequency, pulsed sounds produced bilateral asymmetries in forewing direct depressor muscles (M97, 98, 99) that were positively correlated with asymmetric forewing depression and asymmetries in stroke reversal timing. Bilateral asymmetries in hindwing depressor muscles (M127 and M128 but not M129) were positively correlated with asymmetric hindwing depression and asymmetries in the timing of the hindwing stroke reversal; M129 was negatively correlated with these shifts. Hindwing depressor asymmetries and wing kinematic changes were smaller and shifted in opposite direction than corresponding measurements of the forewings. These findings suggest that intentional steering manoeuvres employ bulk shifts in depressor muscle timing that affect the timing of the stroke reversals thereby establishing asymmetric wing depression. Finally, we found indications that locusts may actively control the timing of forewing rotation and speculate this may be a mechanism for generating steering torques. These effects would act in concert with forces generated by asymmetric wing depression and angle of attack to establish rapid changes in direction.

Acoustic Stimulation↗

Butterfly wing pattern mutants: developmental heterochrony and co-ordinately regulated phenotypes.

Butterfly wings are colored late in development, when pigments are synthesized in specialized wing scale cells in a fixed developmental succession. In this succession, colored pigments are deposited first and the remaining areas are later melanized black or brown. Here we studied the developmental changes underlying two wing pattern mutants, firstly melanic mutants of the swallowtail Papilio glaucus, in which the yellow background is turned black, and secondly a Spotty mutant of the satyrid Bicyclus anynana, which carries two additional eyespots. Despite the very different pattern changes in these two mutants, they are both associated with changes in rates of scale development and correspondingly, the final color pattern. In the melanic swallowtail, background scales originally destined to become yellow (normally developing early and synthesizing papiliochrome) show delayed development, fail to make papiliochrome, and subsequently melanize at the same time as scales in the wild-type black pattern. In the B. anynana eyespot, scale maturation begins with the central white focus, then progresses to the surrounding gold ring and later finishes with melanization of the black center. Mutants showing additional eyespots display accelerated rates of scale development (corresponding to new eyespots) in wing cells not normally occupied by eyespots. Thus by either delaying or accelerating rates of scale development, the final color, or position, of a wing pattern element can be changed. We propose that this heterochrony of scale development is a basic mechanism of color pattern formation on which developmental mutants act to change lepidopteran color patterns.

Animals↗

Quantitative developmental genetic analysis reveals that the ancestral dipteran wing vein prepattern is conserved in Drosophila melanogaster.

Quantitative complementation tests provide a quick test of the hypothesis that a particular gene contributes to segregating phenotypic variation. A set of wild-type alleles is assayed for variation in their ability to complement the degree of dominance of the quantitative effect of a loss of function allele. Analysis of 15 loci known to be involved in wing patterning in Drosophila melanogaster suggests that the genes decapentaplegic, thickveins, EGFR, argos and hedgehog, each of which are involved in secreted growth factor signaling, may contribute to wing shape variation. The phenotype of one deficiency, Df(2R)Px2, which removes blistered/Plexate, is also highly sensitive to the wild-type genetic background and at intermediate expressivity reveals six ectopic veins. These form in the same locations as a projection of the ancestral pattern of dipteran wing veins on- to the D. melanogaster wing. This atavistic phenotype indicates that the wing vein prepatterning mechanism can be conserved in highly derived species, and implies that homoplasic venation patterns may be produced by derepression of vein primordia.

Animals↗