Evaluating whole genome sequence data from the Genetic Absence Epilepsy Rat from Strasbourg and its related non-epileptic strain

PLoS One. 2017 Jul 14;12(7):e0179924. doi: 10.1371/journal.pone.0179924. eCollection 2017.

Abstract

Objective: The Genetic Absence Epilepsy Rats from Strasbourg (GAERS) are an inbreed Wistar rat strain widely used as a model of genetic generalised epilepsy with absence seizures. As in humans, the genetic architecture that results in genetic generalized epilepsy in GAERS is poorly understood. Here we present the strain-specific variants found among the epileptic GAERS and their related Non-Epileptic Control (NEC) strain. The GAERS and NEC represent a powerful opportunity to identify neurobiological factors that are associated with the genetic generalised epilepsy phenotype.

Methods: We performed whole genome sequencing on adult epileptic GAERS and adult NEC rats, a strain derived from the same original Wistar colony. We also generated whole genome sequencing on four double-crossed (GAERS with NEC) F2 selected for high-seizing (n = 2) and non-seizing (n = 2) phenotypes.

Results: Specific to the GAERS genome, we identified 1.12 million single nucleotide variants, 296.5K short insertion-deletions, and 354 putative copy number variants that result in complete or partial loss/duplication of 41 genes. Of the GAERS-specific variants that met high quality criteria, 25 are annotated as stop codon gain/loss, 56 as putative essential splice sites, and 56 indels are predicted to result in a frameshift. Subsequent screening against the two F2 progeny sequenced for having the highest and two F2 progeny for having the lowest seizure burden identified only the selected Cacna1h GAERS-private protein-coding variant as exclusively co-segregating with the two high-seizing F2 rats.

Significance: This study highlights an approach for using whole genome sequencing to narrow down to a manageable candidate list of genetic variants in a complex genetic epilepsy animal model, and suggests utility of this sequencing design to investigate other spontaneously occurring animal models of human disease.

MeSH terms

  • Animals
  • Brain / diagnostic imaging
  • Brain / metabolism
  • Brain / pathology
  • Calcium Channels, T-Type / genetics*
  • DNA / chemistry
  • DNA / isolation & purification
  • DNA / metabolism
  • Disease Models, Animal
  • Electroencephalography
  • Epilepsy, Absence / genetics*
  • Epilepsy, Absence / pathology
  • Female
  • Genome*
  • Genotype
  • High-Throughput Nucleotide Sequencing
  • Male
  • Polymorphism, Single Nucleotide
  • Rats
  • Rats, Wistar
  • Sequence Analysis, DNA

Substances

  • Cacna1h protein, rat
  • Calcium Channels, T-Type
  • DNA

Grants and funding

Pablo M. Casillas-Espinosa was funded by a Melbourne Research Scholarship. Work for this project was supported in part by a NH&MRC Project Grant to Terence O’Brien and Kim Powell and a R.D Wright Career Development Fellowship (1126877) to SP. The current affiliation of JMM, BD technologies, provides support in the form of salaries for JMM, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.