title: Development of a Drosophila seizure model for in vivo high-throughput drug screening.
abstract: An important application of model organisms in neurological research has been to identify and characterise therapeutic approaches for epilepsy, a recurrent seizure disorder that affects > 1% of the human population. Proconvulsant-treated rodent models have been widely used for antiepileptic drug discovery and development, but are not suitable for high-throughput screening. To generate a genetically tractable model that would be suitable for large-scale, high-throughput screening for antiepileptic drug candidates, we characterized a Drosophila chemical treatment model using the GABA(A) receptor antagonist picrotoxin. This proconvulsant, delivered to Drosophila larvae via simple feeding methods suitable for automated screening, generated robust generalised seizures with lethality occurring at doses between 0.3 and 0.5 mg/mL. Electrophysiological analysis of CNS motor neuron output in picrotoxin-treated larvae revealed generalised seizures within minutes of drug exposure. At subthreshold doses for seizure induction, picrotoxin produced an increased frequency of motor neuron action potential bursting, indicating that CNS GABAergic transmission regulates patterned activity. Mutants in the Drosophila Rdl GABA(A) receptor are resistant to picrotoxin, confirming that seizure induction occurs via a conserved GABA(A) receptor pathway. To validate the usefulness of this model for in vivo drug screening, we identified several classes of neuroactive antiepileptic compounds in a pilot screen, including phenytoin and nifedipine, which can rescue the seizures and lethal neurotoxicity induced by picrotoxin. The well-defined actions of picrotoxin in Drosophila and the ease with which compounds can be assayed for antiseizure activity makes this genetically tractable model attractive for high-throughput in vivo screens to identify novel anticonvulsants and seizure susceptibility loci.
authors.affiliation: Cambria Biosciences, 8A Henshaw St., Woburn, MA 01801, USA. gstilwell@cambriabio.com
authors.forename: Geoff E
authors.initials: GE
authors.lastname: Stilwell
authors.forename: Sudipta
authors.initials: S
authors.lastname: Saraswati
authors.forename: J Troy
authors.initials: JT
authors.lastname: Littleton
authors.forename: Scott W
authors.initials: SW
authors.lastname: Chouinard
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000927:Anticonvulsants / Q000494:pharmacology*; D001522:Behavior, Animal / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival; D003292:Convulsants; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004330:Drosophila; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004594:Electrophysiology; D018756:GABA Antagonists; D058787:GABA-A Receptor Antagonists; D007814:Larva; D010433:Pentylenetetrazole; D010641:Phenotype; D010672:Phenytoin / Q000494:pharmacology; D010852:Picrotoxin; D012640:Seizures / Q000139:chemically induced* / Q000188:drug therapy* / Q000235:genetics
chemical_list: D000927:Anticonvulsants; D003292:Convulsants; D018756:GABA Antagonists; D058787:GABA-A Receptor Antagonists; D010433:Pentylenetetrazole; D010672:Phenytoin; D010852:Picrotoxin