Quantitative imaging of morphogen gradients in drosophila imaginal discs Journal Article


Author(s): Kicheva, Anna; Holtzer, Laurent; Wartlick, Ortrud; Schmidt, Thomas S B; González-Gaitán, Marcos A
Article Title: Quantitative imaging of morphogen gradients in drosophila imaginal discs
Affiliation
Abstract: Cells at different positions in a developing tissue receive different concentrations of signaling molecules, called morphogens, and this influences their cell fate. Morphogen concentration gradients have been proposed to control patterning as well as growth in many developing tissues. Some outstanding questions about tissue patterning by morphogen gradients are the following: What are the mechanisms that regulate gradient formation and shape? Is the positional information encoded in the gradient sufficiently precise to determine the positions of target gene domain boundaries? What are the temporal dynamics of gradients and how do they relate to patterning and growth? These questions are inherently quantitative in nature and addressing them requires measuring morphogen concentrations in cells, levels of downstream signaling activity, and kinetics of morphogen transport. Here we first present methods for quantifying morphogen gradient shape in which the measurements can be calibrated to reflect actual morphogen concentrations. We then discuss using fluorescence recovery after photobleaching to study the kinetics of morphogen transport at the tissue level. Finally, we present particle tracking as a method to study morphogen intracellular trafficking.
Keywords: Kinetics; Drosophila; Quantitative Analysis; Morphogen; Image processing; cell migration; cellular distribution; Endosome; Confocal microscopy; Fluorescence; prenatal development; autofluorescence; bleaching; calibration; confocal laser microscopy; diffusion coefficient; fluorescence recovery after photobleaching; light damage; normal distribution; quantitative imaging; Rotavirus; virus particle; developmental biology; histology; methodology
Journal Title: Cold Spring Harbor Protocols
Volume: 8
Issue 5
ISSN: 15596095
Publisher: Cold Spring Harbor Laboratory Press  
Date Published: 2013-05-01
Start Page: 387
End Page: 403
DOI: 10.1101/pdb.top074237
Open access: no
IST Austria Authors
  1. Anna Kicheva
    19 Kicheva
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