Diseases of cortical malformation cause approximately 25% of all cases of epilepsy. They are also the most common cause for surgical resection of epileptic brain tissue. Almost 80% of people with a cortical malformation suffer from epilepsy and greater than 70% of those people have seizures which are not managed by anti-epileptic drugs. Novel treatment strategies are urgently needed to treat this problematic group of epilepsies. In this proposal we will address the hypothesis that loss of astrocytic glutamate reuptake during the development of a cortical malformation acutely disrupts glutamate homeostasis and has long term effects on synaptic connectivity and cortical network function. Normally, astrocytes remove the neurotransmitter glutamate via glutamate transporters. In diseases of cortical malformation, however, astrocytes become reactive which we believe decreases their ability to remove extracellular glutamate. In the developing cortex glutamate directly drives synapse formation. Therefore, we hypothesize that loss of astrocytic glutamate reuptake during the development of a cortical malformation increases extracellular glutamate levels which promotes excitatory synapse formation and leads to long term cortical hyperexcitability. We will test our hypothesis utilizing cutting-edge imaging techniques, electrophysiological recording from astrocytes, in vivo assays of cortical excitability and molecular disruption and augmentation of astrocyte glutamate transport. Our experiments are extremely innovative. We have developed a novel rodent model of cortical malformation which closely replicates focal cortical dysplasia type 1, a disease with no current animal model. We will utilize exciting, novel glutamate biosensor imaging techniques to assay network function and astrocytic glutamate reuptake. We will record cortical glutamate transporter currents, which have not previously been investigated, and we will do so in the malformed cortex. We will utilize laser-scanning photostimulation to spatially map how astrocytic glutamate reuptake is altered in the malformed cortex. Utilizing molecular modulation of astrocytic glutamate transport we will test whether developmental loss of astrocytic glutamate transport is sufficient to induce cortical hyperexcitability and whether increasing glutamate reuptake in the malformed cortex interrupts epileptogenic processes which we believe lead to later network dysfunction. Importantly, we will utilize drugs which are already clinically available to increase glutamate reuptake. Should this approach successfully attenuate cortical hyperexcitability it could be rapidly translated into a potential anti-epileptogenic clinical tool.