PUBLICATION

A Smart Imaging Workflow for Organ-Specific Screening in a Cystic Kidney Zebrafish Disease Model

Authors
Pandey, G., Westhoff, J.H., Schaefer, F., Gehrig, J.
ID
ZDB-PUB-190317-1
Date
2019
Source
International Journal of Molecular Sciences   20(6): (Journal)
Registered Authors
Gehrig, Jochen
Keywords
automated imaging, high-content screening, image analysis, zebrafish
MeSH Terms
  • Polycystic Kidney Diseases/diagnostic imaging*
  • Polycystic Kidney Diseases/metabolism
  • Disease Models, Animal
  • Software
  • Algorithms
  • Kidney/diagnostic imaging*
  • Kidney/metabolism
  • Phenotype
  • Embryo, Nonmammalian/metabolism
  • Humans
  • Workflow
  • Zebrafish
  • Gene Knockdown Techniques
  • Animals
  • Carrier Proteins/genetics*
  • Organ Specificity
  • Small Molecule Libraries/administration & dosage*
  • Small Molecule Libraries/metabolism
  • Image Processing, Computer-Assisted/methods*
(all 19)
PubMed
30875791 Full text @ Int. J. Mol. Sci.
Abstract
The zebrafish is being increasingly used in biomedical research and drug discovery to conduct large-scale compound screening. However, there is a lack of accessible methodologies to enable automated imaging and scoring of tissue-specific phenotypes at enhanced resolution. Here, we present the development of an automated imaging pipeline to identify chemical modifiers of glomerular cyst formation in a zebrafish model for human cystic kidney disease. Morpholino-mediated knockdown of intraflagellar transport protein Ift172 in Tg(wt1b:EGFP) embryos was used to induce large glomerular cysts representing a robustly scorable phenotypic readout. Compound-treated embryos were consistently aligned within the cavities of agarose-filled microplates. By interfacing feature detection algorithms with automated microscopy, a smart imaging workflow for detection, centring and zooming in on regions of interests was established, which enabled the automated capturing of standardised higher resolution datasets of pronephric areas. High-content screening datasets were processed and analysed using custom-developed heuristic algorithms implemented in common open-source image analysis software. The workflow enables highly efficient profiling of entire compound libraries and scoring of kidney-specific morphological phenotypes in thousands of zebrafish embryos. The demonstrated toolset covers all the aspects of a complex whole organism screening assay and can be adapted to other organs, specimens or applications.
Genes / Markers
Figures
Figure Gallery (3 images)
Show all Figures
Expression
Phenotype
Fish Conditions Stage Phenotype Figure
li1Tg + MO3-ift172controlProtruding-mouth
li1Tg + MO3-ift172controlProtruding-mouth
li1Tg + MO3-ift172controlProtruding-mouth
li1Tg + MO3-ift172controlProtruding-mouth
li1Tg + MO3-ift172controlProtruding-mouth
li1Tg + MO3-ift172controlProtruding-mouth
li1Tg + MO3-ift172chemical treatment by environment: sirolimusProtruding-mouth
li1Tg + MO3-ift172chemical treatment by environment: sirolimusProtruding-mouth
li1Tg + MO3-ift172chemical treatment by environment: sirolimusProtruding-mouth
1 - 9 of 9
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
li1TgTransgenic Insertion
    1 - 1 of 1
    Show
    Human Disease / Model
    Human Disease Fish Conditions Evidence
    cystic kidney diseaseli1Tg + MO3-ift172controlTAS
    1 - 1 of 1
    Show
    Sequence Targeting Reagents
    Target Reagent Reagent Type
    ift172MO3-ift172MRPHLNO
    1 - 1 of 1
    Show
    Fish
    Fish
    li1Tg
    li1Tg + MO3-ift172
    1 - 2 of 2
    Show
    Antibodies
    No data available
    Orthology
    No data available
    Engineered Foreign Genes
    Marker Marker Type Name
    EGFPEFGEGFP
    1 - 1 of 1
    Show
    Mapping
    No data available
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    Citation
    Pandey, G., Westhoff, J.H., Schaefer, F., Gehrig, J. (2019) A Smart Imaging Workflow for Organ-Specific Screening in a Cystic Kidney Zebrafish Disease Model. International Journal of Molecular Sciences. 20(6).