Rats deficient in the GAD65 isoform exhibit epilepsy and premature lethality

GABA is synthesized by glutamate decarboxylase (GAD), which has two isoforms, namely, GAD65 and GAD67, encoded by the Gad2 and Gad1 genes, respectively. GAD65-deficient (Gad2(-/-)) mice exhibit a reduction in brain GABA content after 1 month of age and show spontaneous seizures in adulthood. Approximately 25% of Gad2(-/-) mice died by 6 months of age. Our Western blot analysis demonstrated that the protein expression ratio of GAD65 to GAD67 in the brain was greater in rats than in mice during postnatal development, suggesting that the contribution of each GAD isoform to GABA functions differs between these two species. To evaluate whether GAD65 deficiency causes different phenotypes between rats and mice, we generated Gad2(-/-) rats using TALEN genome editing technology. Western blot and immunohistochemical analyses with new antibodies demonstrated that the GAD65 protein was undetectable in the Gad2(-/-) rat brain. Gad2(-/-) pups exhibited spontaneous seizures and paroxysmal discharge in EEG at postnatal weeks 3-4. More than 80% of the Gad2(-/-) rats died at postnatal days (PNDs) 17-23. GABA content in Gad2(-/-) brains was significantly lower than those in Gad2(+/-) and Gad2(+/+) brains at PND17-19. These results suggest that the low levels of brain GABA content in Gad2(-/-) rats may lead to epilepsy followed by premature death, and that Gad2(-/-) rats are more severely affected than Gad2(-/-) mice. Considering that the GAD65/GAD67 ratio in human brains is more similar to that in rat brains than in mouse brains, Gad2(-/-) rats would be useful for further investigating the roles of GAD65 in vivo.

Automated summary

GAD65 deficiency causes different phenotypes between rats and mice, with Gad2(-/-) rats being more severely affected and exhibiting more severe symptoms such as epilepsy and premature death. Considering the similarity of GAD65/GAD67 ratio in human brains to that in rat brains, Gad2(-/-) rats would be useful for further investigating the roles of GAD65 in vivo.