A high-throughput template for optimizing<i>Drosophila</i>organ culture with response-surface methods

Zartman, Jeremiah J.; Restrepo, Simon; Basler, Konrad

doi:10.1242/dev.088872

Cited by 76 publications

(88 citation statements)

References 48 publications

(46 reference statements)

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“…Third instar wing discs were dissected in WM1 medium (Zartman et al, 2013) and placed in a chamber formed by double-sided 3M tape mounted on a glass slide and sealed with a coverslip. Live imaging was performed with a Nikon spinning-disc confocal microscope.…”

Section: Drosophila Wing Disc Culture and Imagingmentioning

confidence: 99%

Essential basal cytonemes take up Hedgehog in the Drosophila wing imaginal disc

et al. 2017

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Morphogen concentration gradients that extend across developmental fields form by dispersion from source cells. In the Drosophila wing disc, Hedgehog (Hh) produced by posterior compartment cells distributes in a concentration gradient to adjacent cells of the anterior compartment. We monitored Hh:GFP after pulsed expression, and analyzed the movement and colocalization of Hh, Patched (Ptc) and Smoothened (Smo) proteins tagged with GFP or mCherry and expressed at physiological levels from bacterial artificial chromosome transgenes. Hh:GFP moved to basal subcellular locations prior to release from posterior compartment cells that express it, and was taken up by basal cytonemes that extend to the source cells. Hh and Ptc were present in puncta that moved along the basal cytonemes and formed characteristic apical-basal distributions in the anterior compartment cells. The basal cytonemes required diaphanous, SCAR, Neuroglian and Synaptobrevin, and both the Hh gradient and Hh signaling declined under conditions in which the cytonemes were compromised. These findings show that in the wing disc, Hh distributions and signaling are dependent upon basal release and uptake, and on cytoneme-mediated movement. No evidence for apical dispersion was obtained.

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Section: Drosophila Wing Disc Culture and Imagingmentioning

confidence: 99%

Essential basal cytonemes take up Hedgehog in the Drosophila wing imaginal disc

et al. 2017

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“…5J,K, wing discs of late third instar larvae were cultured as described previously (Zartman et al, 2013). Images were acquired on a Leica SP5 MP inverse confocal microscope with a 40×/1.25 NA oil immersion objective.…”

Section: Culturing Of Wing Discs and Analysis Of T1 Transitionsmentioning

confidence: 99%

A local difference in Hedgehog signal transduction increases mechanical cell bond tension and biases cell intercalations along the Drosophila anteroposterior compartment boundary

et al. 2015

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Tissue organization requires the interplay between biochemical signaling and cellular force generation. The formation of straight boundaries separating cells with different fates into compartments is important for growth and patterning during tissue development. In the developing Drosophila wing disc, maintenance of the straight anteroposterior (AP) compartment boundary involves a local increase in mechanical tension at cell bonds along the boundary. The biochemical signals that regulate mechanical tension along the AP boundary, however, remain unknown. Here, we show that a local difference in Hedgehog signal transduction activity between anterior and posterior cells is necessary and sufficient to increase mechanical tension along the AP boundary. This difference in Hedgehog signal transduction is also required to bias cell rearrangements during cell intercalations to keep the characteristic straight shape of the AP boundary. Moreover, severing cell bonds along the AP boundary does not reduce tension at neighboring bonds, implying that active mechanical tension is upregulated, cell bond by cell bond. Finally, differences in the expression of the homeodomain-containing protein Engrailed also contribute to the straight shape of the AP boundary, independently of Hedgehog signal transduction and without modulating cell bond tension. Our data reveal a novel link between local differences in Hedgehog signal transduction and a local increase in active mechanical tension of cell bonds that biases junctional rearrangements. The large-scale shape of the AP boundary thus emerges from biochemical signals inducing patterns of active tension on cell bonds.

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“…For a biophysical application of ER reconstructions we refer to Sbalzarini et al (2005Sbalzarini et al ( , 2006. The Drosophila wing data come from Zartman et al (2013). Figure 6a,d show the maximum-intensity projections of the raw images 16 of the ER (325 × 250 × 16 pixels) and the wing disc (512×512×12 pixels), respectively.…”

Section: D Segmentation Of Confocal Microscopy Datamentioning

confidence: 99%

Coupling Image Restoration and Segmentation: A Generalized Linear Model/Bregman Perspective

2013

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We introduce a new class of data-fitting energies that couple image segmentation with image restoration. These functionals model the image intensity using the statistical framework of generalized linear models. By duality, we establish an information-theoretic interpretation using Bregman divergences. We demonstrate how this formulation couples in a principled way image restoration tasks such as denoising, deblurring (deconvolution), and inpainting with segmentation. We present an alternating minimization algorithm to solve the resulting composite photometric/geometric inverse problem. We use Fisher scoring to solve the photometric problem and to provide asymptotic uncertainty estimates. We derive the shape gradient of our data-fitting energy and investigate convex relaxation for the geometric problem. We introduce a new alternating split-Bregman strategy to

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A high-throughput template for optimizingDrosophilaorgan culture with response-surface methods

Cited by 76 publications

References 48 publications

Essential basal cytonemes take up Hedgehog in the Drosophila wing imaginal disc

Essential basal cytonemes take up Hedgehog in the Drosophila wing imaginal disc

A local difference in Hedgehog signal transduction increases mechanical cell bond tension and biases cell intercalations along the Drosophila anteroposterior compartment boundary

Coupling Image Restoration and Segmentation: A Generalized Linear Model/Bregman Perspective

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