ZeTox – Toxicology Services

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icon-zeglobaltox ZeGlobalTox – 3 Analysis Into a 1 Procedure

ZeGlobalTox is a novel experimental procedure that integrates the analysis of cardio-, neuro- and hepatotoxicity into a single procedure. The three sequential assays are performed in the same larvae, allowing the reduction of animals and streamlining the experimental pipeline. The high predictivity of ZeGlobalTox allows reducing the use of mammals in organ-toxicity screening.

Advantages

Simultaneous assessment of NeuroTox, CardioTox and HepatoTox.

High predictivity referred to humans: 89% specificity, 68% sensitivity and 78% accuracy.

Animal testing and drug volumes reducement.

Method description

A zebrafish transgenic line expressing green fluorescent protein (GFP) in cardiomyocytes and red fluorescent protein (RFP) in hepatocytes is used to determine cardiotoxicity, neurotoxicity, and hepatotoxicity within the experimental window 120 to 128 hpf.

Readouts

  • Neurotoxicity at 123 hpf
    • Locomotor activity
    • Thigmotaxis
    • Altered response to light stimuli
    • Habituation
    • Seizures
  • Cardiotoxicity at 124 hpf
    • Heart beat frequency (ventricle and atrium)
    • Arrhythmias
    • Ejection fraction
    • QT prolongation
    • Fibrillation
    • Cardiac arrest
  • Hepatotoxicity at 128 hpf
    • Liver area: liver necrosis and hepatomegaly
    • Yolk lipid retention
    • Steatosis: fat accumulation in liver
Figure 1. Larval videotracking of basal locomotion activity acquired by NOLDUS system. Red lines show larval movements.
ZeTox_Neurotox_Figure3_-Locomotion
Figure 2.  General motor activity assessment. Light/Dark locomotion pattern of zebrafish larvae in response to a negative control, to a neurotoxic compound (positive control) and a study compound that doesn’t produce neurotoxic effect on basal locomotor activity.
Figure 3. Heart tomography of 3dfp transgenic zebrafish larva labelling cardiomyocytes with GFP.
Figure 4. Yolk lipid retention evaluation. (A) Proportion of zebrafish larvae that show yolk lipid retention after treatment with a molecule of interest compared to negative and positive controls. (B) Representative images of zebrafish after specific fat staining showing a healthy larva (image on the right), larva with high amount of yolk lipid retention (middle) and larva with few amount of lipid retention (left).

References

  1. Cornet C, Calzolari S, Miñana-Prieto R, Dyballa S, van Doornmalen E, Rutjes H, Savy T, D'Amico D, Terriente J. ZeGlobalTox: An Innovative Approach to Address Organ Drug Toxicity Using Zebrafish. Int J Mol Sci. 2017 Apr 19;18(4):864.