ZeGenesis

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Neurophenotyping Analysis

Central Nervous System (CNS) functions are highly conserved. Behavioral responses resemble mammalian species and have robust, validated patterns.
Zebrafish allow  simultaneous, non-invasive, in vivo characterization of behavioral alterations inside well-plates, thanks to robotic tracking systems associated to putative neuroprotective/neurotoxic molecules or genes. 

Current readouts

Analysis

Analysis

  • Angle turns
  • Total Velocity
  • Max Velocity
  • Center vs. Periphery
  • Total distance
  • etc.

Assays

General motor activity and visual response assay

Total distance moved, locomotion pattern and visual stimuli response.

Natural locomotion behaviour of zebrafish larvae is characterized by high activity in darkness and immobility in light environment. 

Data shows difference in larvae activity exposed to alternating dark and light environment by treatment with sedative (Clemizole hydrochloride) and control siblings (DMSO). 

Figure: Light/dark behavioral response
Thigmotaxis assay

Evaluation of the anxiety state of the larvae after putative anxiogenic or anxiolytic drug exposure or genetic change. 

Total distance moved and the time spent in either centre or periphery of the well plate. 

Data is measured and compared to assess putative anxiolytic or anxiogenic effects of the tested compounds. 

Figure: Evaluation of the anxiety state
Habituation assay

Habituation is the capacity to gradually reduce the response to external and repeated stimuli. It is evaluated to identify short memory and learning process defects caused by the tested compound/gene. 

Total distance moved, locomotion activity and visual stimuli response.

Neuroactive compounds and anxiogenics (EtOH) cause alterations of the learning processes and larvae do not “habituate” to the stimulus. 

Anxiolytics (Phenprobamate) and sedatives decrease general locomotion and reduce rapidly the response to repeated stimuli

Total distance moved
Convulsive assay

Seizures are characterized by a fast-darting and erratic movements, detected as an increase in the maximum velocity and the number of angle turns of the larvae in response to light flash. 

MaxVelocity and angle Turn

The neurotoxin Pentyleneterazole produces seizures that are reverted by the anti-convulsive compounds Topiramate and Valproic Acid.

DMSO PTZ

Specific neuronal population analysis

Specific neuronal cell lines can be analyzed in zebrafish transgenic neuronal reporter lines. This cellular readout might be helpful to identify neuronal survival, axonal defects and deeper understanding on the mechanism of action of tested compounds or genes.

Tg(isl1:GFP) line specifically labels motorneurons allowing to study effects on this cell population