Emerging evidence suggest a causal role of P2X7 in seizure-induced pathology and epilepsy in adults. New data now also suggests a role of purinergic signalling, in particular P2X7, during seizures in neonates. There are, however, several important unanswered questions. These include elucidating the use of P2X7-targeting drugs and determine impact of clinical outcomes. My project will use cutting edge, novel techniques to elucidate the role of P2X7 related to neonate seizures, using pre-clinical mouse models of seizures in neonates, highly specific P2X7 antagonists from our industrial collaborator J&R. While ATP is widely assumed to be released during increased neuronal activity, this has never been demonstrated in vivo. We now have access to a novel technology, capable of recording concentrations of ATP at nanomolar concentrations in situ and, more importantly, a much higher temporal resolution than microdialysis. Pilot studies in the host laboratory have indicated a spike in ATP release during experimental seizures. The GFP-P2X7 reporter mouse we will use will help resolve some of the controversies regarding both constitutive expression in the CNS during development and changes in expression in response to an insult. Secondly, conditional, cell-specific P2X7 KO mice allow for the dissection of the contribution of P2X7 from neurons and microglia to acute pathology (e.g. seizures) and clinical outcomes.
