Epilepsy in kcnj10 morphant zebrafish assessed with a novel method for long-term EEG recordings

PLoS One. 2013 Nov 14;8(11):e79765. doi: 10.1371/journal.pone.0079765. eCollection 2013.

Abstract

We aimed to develop and validate a reliable method for stable long-term recordings of EEG activity in zebrafish, which is less prone to artifacts than current invasive techniques. EEG activity was recorded with a blunt electrolyte-filled glass pipette placed on the zebrafish head mimicking surface EEG technology in man. In addition, paralysis of agarose-embedded fish using D-tubocurarine excluded movement artifacts associated with epileptic activity. This non-invasive recording technique allowed recordings for up to one hour and produced less artifacts than impaling the zebrafish optic tectum with a patch pipette. Paralyzed fish survived, and normal heartbeat could be monitored for over 1h. Our technique allowed the demonstration of specific epileptic activity in kcnj10a morphant fish (a model for EAST syndrome) closely resembling epileptic activity induced by pentylenetetrazol. This new method documented that seizures in the zebrafish EAST model were ameliorated by pentobarbitone, but not diazepam, validating its usefulness. In conclusion, non-invasive recordings in paralyzed EAST syndrome zebrafish proved stable, reliable and robust, showing qualitatively similar frequency spectra to those obtained from pentylenetetrazol-treated fish. This technique may prove particularly useful in zebrafish epilepsy models that show infrequent or conditional seizure activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anticonvulsants / administration & dosage
  • Anticonvulsants / pharmacology
  • Brain / drug effects
  • Brain / physiopathology
  • Convulsants / adverse effects
  • Diazepam / administration & dosage
  • Diazepam / pharmacology
  • Disease Models, Animal
  • Electroencephalography*
  • Epilepsy / drug therapy
  • Epilepsy / genetics*
  • Epilepsy / physiopathology*
  • Gene Knockdown Techniques
  • Pentylenetetrazole / adverse effects
  • Potassium Channels, Inwardly Rectifying / genetics*
  • Seizures / chemically induced
  • Seizures / drug therapy
  • Seizures / genetics
  • Seizures / physiopathology
  • Zebrafish*

Substances

  • Anticonvulsants
  • Convulsants
  • Kcnj10 (channel)
  • Potassium Channels, Inwardly Rectifying
  • Diazepam
  • Pentylenetetrazole

Grant support

UCL work was supported by grants from the Peter Samuel Trust Fund (grant number 939) (to AAZ), the David and Elaine Potter Charitable Foundation and Grocers’ Charity (to RK), St Peter’s Trust for Kidney, Bladder and Prostate Research (to AAZ, DB and RK), Kids Kidney Research and Garfield Weston Foundation (to DB and RK), and by the EU FP7 program (EURenOmics grant agreement number 305608 (to DB, RK)). Funding for FM was provided via a doctoral training grant from the Royal Veterinary College (RVC) (to CR). Funding for consumables and animals at the RVC (to CR and FM) was provided by a RVC start-up grant (to CR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.