The distribution of PS integrins, laminin A and F-actin during key stages in Drosophila wing development

Development ◽  
1993 ◽  
Vol 117 (2) ◽  
pp. 509-523 ◽  
Author(s):  
D. Fristrom ◽  
M. Wilcox ◽  
J. Fristrom
Keyword(s):  
Wing Disc ◽  
Wing Development ◽  
Filamentous Actin ◽  
Pupal Development ◽  
Drosophila Wing ◽  
Junction Formation ◽  
Hair Morphogenesis ◽  
Cytoplasmic Processes ◽  
Intercellular Connections

We first summarize wing development during metamorphosis of Drosophila and identify four critical steps in the conversion of a folded single layered wing disc to a flat bilayered wing. Each step occurs twice, once during the 12 hour prepupal period and again during the 84 hour pupal period. (1) Apposition in which basal surfaces of dorsal and ventral epithelia come close together. (2) Adhesion in which basal junctions form between the apposed basal surfaces. (3) Expansion in which wing area increases as a result of cells flattening. (4) Separation in which dorsal and ventral epithelia are separated by a bulky extracellular matrix but remain connected by slender cytoplasmic processes containing the microtubules and microfilaments of the transalar cytoskeleton. Disc ultrastructure is correlated with the distribution of the beta chain of integrin, laminin A, and filamentous actin for each key stage of pupal development. Integrin and laminin exhibit a mutually exclusive distribution from the adhesion stage onwards. Integrin is present on the basal surface of intervein cells but not on vein cells whereas laminin A is absent from the basal surfaces of intervein cells but is present on vein cells. We conclude that laminin is not a ligand for integrin in this context. During apposition and adhesion stages integrin is broadly distributed over the basal and lateral surfaces of intervein cells but subsequently becomes localized to small basal foci. These foci correspond to basal contact zones between transalar processes. The distribution of filamentous actin is dynamic, changing from an apical distribution during hair morphogenesis to a basal distribution as the transalar cytoskeleton develops. Basal adherens-type junctions are first evident during the adhesion stage and become closely associated with the transalar cytoskeleton during the separation stage. Thus, basal junction formation occurs in two discrete steps; intercellular connections are established first and junction/cytoskeletal connections are formed about 20 hours later. These observations provide a basis for future investigations of integrin mediated adhesion in vivo.

scholarly journals In vivo relevance of intercellular calcium signaling in Drosophila wing development

2017 ◽  
Author(s):  
Qinfeng Wu ◽  
Pavel A. Brodskiy ◽  
Francisco Huizar ◽  
Jamison J. Jangula ◽  
Cody Narciso ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Wing Disc ◽  
Chemical Stimulus ◽  
Wing Development ◽  
Dependent Manner ◽  
Drosophila Wing ◽  
Wing Discs ◽  
Vein Patterning ◽  
Dose Dependent

AbstractRecently, organ-scale intercellular Ca2+ transients (ICTs) were reported in the Drosophila wing disc. However, the functional in vivo significance of ICTs remains largely unknown. Here we demonstrate the in vivo relevance of intercellular Ca2+ signaling and its impact on wing development. We report that Ca2+ signaling in vivo decreases as wing discs mature. Ca2+ signaling ex vivo responds to fly extract in a dose-dependent manner. This suggests ICTs occur in vivo due to chemical stimulus that varies in concentration during development. RNAi mediated inhibition of genes required for ICTs results in defects in the size, shape, and vein patterning of adult wings. It also leads to reduction or elimination of in vivo Ca2+ transients. Further, perturbations to the extracellular matrix along the basal side of the wing disc stimulates intercellular Ca2+ waves. This is the first identified chemically defined, non-wounding stimulus of ICTs. Together, these results point toward specific in vivo functions of intercellular Ca2+ signaling to mediate mechanical stress dissipation and ensure robust patterning during development.

Regulation of Apterous activity inDrosophilawing development

Development ◽  
2001 ◽  
Vol 128 (22) ◽  
pp. 4615-4622 ◽  
Author(s):  
Ulrich Weihe ◽  
Marco Milán ◽  
Stephen M. Cohen
Keyword(s):  
Complex Formation ◽  
Wing Development ◽  
Homeodomain Protein ◽  
Activity Levels ◽  
Present Evidence ◽  
Drosophila Wing ◽  
Lim Domains ◽  
Regulation Of Activity ◽  
Lim Homeodomain

Apterous is a LIM-homeodomain protein that confers dorsal compartment identity in Drosophila wing development. Apterous activity requires formation of a complex with a co-factor, Chip/dLDB. Apterous activity is regulated during wing development by dLMO, which competes with Apterous for complex formation. Here, we present evidence that complex formation between Apterous, Chip and DNA stabilizes Apterous protein in vivo. We also report that a difference in the ability of Chip to bind the LIM domains of Apterous and dLMO contributes to regulation of activity levels in vivo.

scholarly journals An Assay to Detect In Vivo Y Chromosome Loss in Drosophila Wing Disc Cells

2012 ◽  
Vol 2 (9) ◽  
pp. 1095-1102 ◽  
Author(s):  
Janos Szabad ◽  
Hugo J. Bellen ◽  
Koen J. T. Venken
Keyword(s):  
Y Chromosome ◽  
Wing Disc ◽  
Chromosome Loss ◽  
Drosophila Wing ◽  
Disc Cells

scholarly journals Microvilli-derived Extracellular Vesicles Govern Morphogenesis in Drosophila wing epithelium

2020 ◽  
Author(s):  
Ilse Hurbain ◽  
Anne-Sophie Macé ◽  
Maryse Romao ◽  
Lucie Sengmanivong ◽  
Laurent Ruel ◽  
...  
Keyword(s):  
Long Range ◽  
Extracellular Vesicles ◽  
Cell Biology ◽  
Extracellular Vesicle ◽  
Wing Disc ◽  
Cellular Mechanisms ◽  
Long Distance ◽  
Drosophila Wing ◽  
And Function

ABSTRACTThe regulation and coordination of developmental processes involves the secretion of morphogens and membrane carriers, including extracellular vesicles, which facilitate their transport over long distance. The long-range activity of the Hedgehog morphogen is conveyed by extracellular vesicles. However, the site and the molecular basis of their biogenesis remains unknown. By combining fluorescence and electron microscopy combined with genetics and cell biology approaches, we investigated the origin and the cellular mechanisms underlying extracellular vesicle biogenesis, and their contribution to Drosophila wing disc development, exploiting Hedgehog as a long-range morphogen. We show that microvilli of Drosophila wing disc epithelium are the site of generation of small extracellular vesicles that transport Hedgehog across the tissue. This process requires the Prominin-like protein, whose activity, together with interacting cytoskeleton components and lipids, is critical for maintaining microvilli integrity and function in secretion. Our results provide the first evidence that microvilli-derived extracellular vesicles contribute to Hedgehog long-range signaling activity highlighting their physiological significance in tissue development in vivo.

scholarly journals Junctional tumor suppressors interact with 14-3-3 proteins to control planar spindle alignment

2019 ◽  
Vol 218 (6) ◽  
pp. 1824-1838 ◽  
Author(s):  
Yu-ichiro Nakajima ◽  
Zachary T. Lee ◽  
Sean A. McKinney ◽  
Selene K. Swanson ◽  
Laurence Florens ◽  
...  
Keyword(s):  
Cell Fate ◽  
Tumor Suppressors ◽  
Mitotic Spindle ◽  
Wing Disc ◽  
Spindle Orientation ◽  
Epithelial Proliferation ◽  
Tissue Architecture ◽  
Drosophila Wing ◽  
Discs Large

Proper orientation of the mitotic spindle is essential for cell fate determination, tissue morphogenesis, and homeostasis. During epithelial proliferation, planar spindle alignment ensures the maintenance of polarized tissue architecture, and aberrant spindle orientation can disrupt epithelial integrity. Nevertheless, in vivo mechanisms that restrict the mitotic spindle to the plane of the epithelium remain poorly understood. Here we show that the junction-localized tumor suppressors Scribbled (Scrib) and Discs large (Dlg) control planar spindle orientation via Mud and 14-3-3 proteins in the Drosophila wing disc epithelium. During mitosis, Scrib is required for the junctional localization of Dlg, and both affect mitotic spindle movements. Using coimmunoprecipitation and mass spectrometry, we identify 14-3-3 proteins as Dlg-interacting partners and further report that loss of 14-3-3s causes both abnormal spindle orientation and disruption of epithelial architecture as a consequence of basal cell delamination and apoptosis. Combined, these biochemical and genetic analyses indicate that 14-3-3s function together with Scrib, Dlg, and Mud during planar cell division.

scholarly journals Arf6 is necessary for senseless expression in response to Wingless signalling during Drosophila wing development

Biology Open ◽  
2021 ◽  
Author(s):  
Julien Marcetteau ◽  
Tamàs Matusek ◽  
Frédéric Luton ◽  
Pascal P. Thérond
Keyword(s):  
Signal Transduction ◽  
Developmental Model ◽  
Wing Development ◽  
Wing Margin ◽  
Drosophila Wing ◽  
Gtp Binding ◽  
Wide Range ◽  
High Level

Wnt signalling is a core pathway involved in a wide range of developmental processes throughout the metazoa. In vitro studies have suggested that the small GTP binding protein Arf6 regulates upstream steps of Wnt transduction, by promoting the phosphorylation of the Wnt co-receptor, LRP6, and the release of β-catenin from the adherens junctions. To assess the relevance of these previous findings in vivo, we analysed the consequence of the absence of Arf6 activity on Drosophila wing patterning, a developmental model of Wnt/Wingless signalling. We observed a dominant loss of wing margin bristles and Senseless expression in Arf6 mutant flies, phenotypes characteristic of a defect in high level Wingless signalling. In contrast to previous findings, we show that Arf6 is required downstream of Armadillo/β-catenin stabilisation in Wingless signal transduction. Our data suggest that Arf6 modulates the activity of a downstream nuclear regulator of Pangolin activity in order to control the induction of high level Wingless signalling. Our findings represent a novel regulatory role for Arf6 in Wingless signalling.

scholarly journals A nanobody-based toolset to investigate the role of protein localization and dispersal in Drosophila

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Stefan Harmansa ◽  
Ilaria Alborelli ◽  
Dimitri Bieli ◽  
Emmanuel Caussinus ◽  
Markus Affolter
Keyword(s):  
Fusion Proteins ◽  
Imaginal Disc ◽  
Protein Localization ◽  
Wing Disc ◽  
Drosophila Wing ◽  
System A ◽  
Polarized Cells ◽  
Lateral Plane

The role of protein localization along the apical-basal axis of polarized cells is difficult to investigate in vivo, partially due to lack of suitable tools. Here, we present the GrabFP system, a collection of four nanobody-based GFP-traps that localize to defined positions along the apical-basal axis. We show that the localization preference of the GrabFP traps can impose a novel localization on GFP-tagged target proteins and results in their controlled mislocalization. These new tools were used to mislocalize transmembrane and cytoplasmic GFP fusion proteins in the Drosophila wing disc epithelium and to investigate the effect of protein mislocalization. Furthermore, we used the GrabFP system as a tool to study the extracellular dispersal of the Decapentaplegic (Dpp) protein and show that the Dpp gradient forming in the lateral plane of the Drosophila wing disc epithelium is essential for patterning of the wing imaginal disc.

Relationships between extramacrochaetae and Notch signalling in Drosophila wing development

Development ◽  
2000 ◽  
Vol 127 (11) ◽  
pp. 2383-2393 ◽  
Author(s):  
A. Baonza ◽  
J.F. de Celis ◽  
A. Garcia-Bellido
Keyword(s):  
Cell Proliferation ◽  
Genetic Interaction ◽  
Wing Disc ◽  
Notch Signalling ◽  
Wing Development ◽  
Notch Activity ◽  
Drosophila Wing ◽  
Vein Formation ◽  
Gene Enhancer ◽  
Enhancer Of Split

The function of extramacrochaetae is required during the development of the Drosophila wing in processes such as cell proliferation and vein differentiation. extramacrochaetae encodes a transcription factor of the HLH family, but unlike other members of this family, Extramacrochaetae lacks the basic region that is involved in interaction with DNA. Some phenotypes caused by extramacrochaetae in the wing are similar to those observed when Notch signalling is compromised. Furthermore, maximal levels of extramacrochaetae expression in the wing disc are restricted to places where Notch activity is higher, suggesting that extramacrochaetae could mediate some aspects of Notch signalling during wing development. We have studied the relationships between extramacrochaetae and Notch in wing development, with emphasis on the processes of vein formation and cell proliferation. We observe strong genetic interaction between extramacrochaetae and different components of the Notch signalling pathway, suggesting a functional relationship between them. We show that the higher level of extramacrochaetae expression coincides with the domain of expression of Notch and its downstream gene Enhancer of split-m(beta). The expression of extramacrochaetae at the dorso/ventral boundary and in boundary cells between veins and interveins depends on Notch activity. We propose that at least during vein differentiation and wing margin formation, extramacrochaetae is regulated by Notch and collaborates with other Notch-downstream genes such as Enhancer of split-m(beta).

Quantitative EM of gap junction distribution in mutant drosophila wing discs

1983 ◽  
Vol 41 ◽  
pp. 720-721
Author(s):  
J.S. Ryerse
Keyword(s):  
Gap Junctions ◽  
Growth Control ◽  
Intercellular Junctions ◽  
Wing Disc ◽  
En Bloc ◽  
Metabolic Cooperation ◽  
Drosophila Wing ◽  
Adult Wing ◽  
Wing Discs ◽  
Wing Pouch

Gap junctions are intercellular junctions found in both vertebrates and invertebrates through which ions and small molecules can pass. Their distribution in tissues could be of critical importance for ionic coupling or metabolic cooperation between cells or for regulating the intracellular movement of growth control and pattern formation factors. Studies of the distribution of gap junctions in mutants which develop abnormally may shed light upon their role in normal development. I report here the distribution of gap junctions in the wing pouch of 3 Drosophila wing disc mutants, vg (vestigial) a cell death mutant, 1(2)gd (lethal giant disc) a pattern abnormality mutant and 1(2)gl (lethal giant larva) a neoplastic mutant and compare these with wildtype wing discs.The wing pouch (the anlagen of the adult wing blade) of a wild-type wing disc is shown in Fig. 1 and consists of columnar cells (Fig. 5) joined by gap junctions (Fig. 6). 14000x EMs of conventionally processed, UA en bloc stained, longitudinally sectioned wing pouches were enlarged to 45000x with a projector and tracings were made on which the lateral plasma membrane (LPM) and gap junctions were marked.

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