Dysregulation of excitatory amino acid transporter 2 (EAAT2) contributes to the development of temporal lobe epilepsy (TLE). Several strategies for increasing total EAAT2 levels have been proposed. However, the mechanism underlying the oligomeric assembly of EAAT2, impairment of which inhibits the formation of functional oligomers by EAAT2 monomers, is still poorly understood. In the present study, we identified E3 ubiquitin ligase AMFR as an EAAT2-interacting protein. AMFR specifically increased the level of EAAT2 oligomers rather than inducing protein degradation through K542-specific ubiquitination. By using tissues from humans with TLE and epilepsy model mice, we observed that AMFR and EAAT2 oligomer levels were simultaneously decreased in the hippocampus. Screening of 2386 FDA-approved drugs revealed that the most common analgesic/antipyretic medicine, acetaminophen (APAP), can induce AMFR transcriptional activation via transcription factor SP1. Administration of APAP protected against pentylenetetrazol-induced epileptogenesis. In mice with chronic epilepsy, APAP treatment partially reduced the occurrence of spontaneous seizures and greatly enhanced the antiepileptic effects of 17AAG, an Hsp90 inhibitor that upregulates total EAAT2 levels, when the 2 compounds were administered together. In summary, our studies reveal an essential role for AMFR in regulating the oligomeric state of EAAT2 and suggest that APAP can improve the efficacy of EAAT2-targeted antiepileptic treatments.

Both the cellular- and population-level properties of involved neurons are essential for unveiling the learning and memory functions of the brain. To give equal attention to these two aspects, neural sensors based on microelectrode arrays (MEAs) have been in the limelight due to their noninvasive detection and regulation capabilities. Here, we fabricated a neural sensor using carboxylated graphene/3,4-ethylenedioxythiophene:polystyrenesulfonate (cGO/PEDOT:PSS), which is effective in sensing and monitoring neuronal electrophysiological activity in vitro for a long time. The cGO/PEDOT:PSS-modified microelectrodes exhibited a lower electrochemical impedance (7.26 ± 0.29 kΩ), higher charge storage capacity (7.53 ± 0.34 mC/cm²), and improved charge injection (3.11 ± 0.25 mC/cm²). In addition, their performance was maintained after 2 to 4 weeks of long-term cell culture and 50,000 stimulation pulses. During neural network training, the sensors were able to induce learning function in hippocampal neurons through precise electrical stimulation and simultaneously detect changes in neural activity at multiple levels. At the cellular level, not only were three kinds of transient responses to electrical stimulation sensed, but electrical stimulation was also found to affect inhibitory neurons more than excitatory neurons. As for the population level, changes in connectivity and firing synchrony were identified. The cGO/PEDOT:PSS-based neural sensor offers an excellent tool in brain function development and neurological disease treatment.