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30 reviewsEsketamine, commonly used to treat treatment-resistant depression, has pharmacological mechanisms that remain incompletelyunderstood. Brain organoids offer a human-relevant platform for investigating the cellular and molecular effects of drugs. In thisstudy, we investigated the effects of esketamine on the electrophysiology and metabolism of brain organoids derived from iPSCsof healthy control subjects and depressed patients. Continuous monitoring revealed that esketamine treatment significantly1234567890();,:decreased both the frequency and amplitude of action potentials, with the most pronounced reduction occurring within 4 h. Highconcentrations (1.5 mg/L) produced a stronger inhibitory effect, while organoids treated with a low concentration (0.25 mg/L)showed a recovery in action potential frequency after one week, although levels remained below pre-treatment values—a recoverynot observed in the high-concentration group. Single-cell RNA sequencing demonstrated that esketamine modulated energymetabolism and induced metabolic reprogramming in a concentration- and time-dependent manner. Furthermore, by inhibitingoxidative phosphorylation and glycolysis separately and assessing cytosolic Ca2+ levels, we found that esketamine may regulateNMDAR activity and electrophysiology through energy metabolism pathways. These findings reveal a potential mechanism foresketamine’s effects and offer new insights for clinical treatment strategies.Molecular Psychiatry;
