Conjugated 3α5β neuroactive steroids, such as allopregnanolone and pregnanolone, have garnered significant interest due to their potential therapeutic effects in various neurological conditions. Previously, the neuroprotective effect of 3α5β pregnanolone glutamate (3α5β-PG) was demonstrated in the model of focal cerebral ischemia in immature rats. The goal of this study was to determine electrophysiological correlates of ischemia-induced seizures and validate effects of 3α5β-PG treatment on seizure’s onset and EEG changes in twelve-day-old (P12) Wistar rats. Focal cerebral ischemia was induced by the infusion of the endothelin-1 (ET-1) into the right dorsal hippocampus in a concentration 40 pmol and total volume 1μl. Electrodes had been implanted contralateral into the dorsal hippocampus and over the left sensorimotor cortex. The 3α5β-PG at the dose 1 mg/kg was administrated intraperitoneally 5 minutes after the end of ET-1 infusion. Animals were then video/EEG-monitored for 120 minutes and parameters of electrographic seizures (frequency, duration, spikes count) as well as pattern of power spectral density (PSD) changes were evaluated. The results revealed that treatment with pregnanolone glutamate led to impairment of ischemia-induced seizures by ~90% significantly reduced total number of spikes by ~60%. Moreover the 3α5β-PG treatment significantly reduced power of delta, theta, alpha, beta, and gamma spectra in comparison with ischemic animals. Our findings have made a substantial contribution to link between pathology in the developing brain and novel therapeutic approaches. The research was supported by project: GAČR 23-05746S.