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Neuroscience JoVE (Open Access) Citable · DOI

Behavioral And Physiological Analysis In A Zebrafish Model Of Epilepsy

DOI: 10.3791/58837-v
What you'll learn
  • ✓ Perform DEPDC5 gene knockdown via microinjection in zebrafish embryos
  • ✓ Measure locomotor activity and seizure-like behavior in larval zebrafish
  • ✓ Conduct electrophysiological recordings from zebrafish larval brain tissue
  • ✓ Characterize epilepsy phenotypes using behavioral and physiological assays
Protocol

Here, we present a protocol for the development and the characterization of a zebrafish model of epilepsy resulting from the transient inhibition of the DEPDC5 gene.

Difficulty
advanced
Total time
~3–4 days (embryo preparation through electrophysiology endpoint)
Model organism
Zebrafish (Danio rerio) larvae
Biosafety
BSL-1

Steps

1
Prepare embryos and perform DEPDC5 microinjection

Collect and stage zebrafish embryos, then perform morpholino or mRNA microinjection targeting DEPDC5 to induce transient gene inhibition and generate the epilepsy model.

▶ 00:28
2
Analyze global locomotor activity patterns

Record and quantify spontaneous and evoked movement behavior in live larvae to detect hyperactivity or seizure-like episodes characteristic of the DEPDC5 knockdown phenotype.

▶ 02:29
3
Perform touch-evoked escape response testing

Apply mechanical stimuli to larvae and measure escape response latency, velocity, and distance traveled to assess sensorimotor function and seizure susceptibility.

▶ 03:48
4
Prepare larvae for electrophysiological recording

Immobilize and mount larval zebrafish, then prepare the exposed brain for field potential recording using appropriate electrode placement and solution exchange.

▶ 04:55
5
Acquire and analyze larval brain field recordings

Record spontaneous or evoked electrical activity from the larval brain and characterize seizure-like discharges, spike frequency, and network oscillations in DEPDC5-deficient animals.

▶ 06:54
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