2-AG-Mediated Control of GABAergic Signaling Is Impaired in a Model of Epilepsy

Repeated seizures result in a persistent maladaptation of endocannabinoid (eCB) signaling, mediated part by anandamide sig-naling deficiency in the basolateral amygdala (BLA) that manifests as aberrant synaptic function and altered emotional behav-ior. Here, we determined the effect of repeated seizures (kindling) on 2-arachidonoylglycerol (2-AG) signaling on GABA transmission by directly measuring tonic and phasic eCB-mediated retrograde signaling in an in vitro BLA slice preparation from male rats. We report that both activity-dependent and muscarinic acetylcholine receptor (mAChR)-mediated depression of GABA synaptic transmission was reduced following repeated seizure activity. These effects were recapitulated in sham rats by preincubating slices with the 2-AG synthesizing enzyme inhibitor DO34. Conversely, preincubating slices with the 2-AG degrading enzyme inhibitor KML29 rescued activity-dependent 2-AG signaling, but not mAChR-mediated synaptic depression, over GABA transmission in kindled rats. These effects were not attributable to a change in cannabinoid type 1 (CB1) receptor sensitivity or altered 2-AG tonic signaling since the application of the highly selective CB1 receptor agonist CP55,940 pro-voked a similar reduction in GABA synaptic activity in both sham and kindled rats, while no effect of either DO34 or of the CB1 inverse agonist AM251 was observed on frequency and amplitude of spontaneous IPSCs in either sham or kindled rats. Collectively, these data provide evidence that repeated amygdala seizures persistently alter phasic 2-AG-mediated retrograde signaling at BLA GABAergic synapses, probably by impairing stimulus-dependent 2-AG synthesis/release, which contributes to the enduring aberrant synaptic plasticity associated with seizure activity.

Automated summary

Repeated seizures result in persistent alteration of endocannabinoid (eCB) signaling, particularly a deficiency in anandamide signaling in the basolateral amygdala (BLA), leading to abnormal synaptic function and emotional behavior. This study found that repeated seizures reduced tonic and phasic eCB-mediated retrograde signaling in GABA transmission. These effects were not due to changes in CB1 receptor sensitivity or altered baseline 2-AG signaling, but rather impaired stimulus-dependent 2-AG synthesis/release.