Sprague-Dawley rat pups at postnatal day (PND) 6 and 15-month-old male C57BL/6 mice were used in the present study. For ketamine GA, rat pups at PND 6 or 15-month-old mice received ketamine (75 mg/kg) intraperitoneally daily for three consecutive days. For sevoflurane GA, animals were put in an anaesthetizing chamber delivered with 3% sevoflurane plus 30% oxygen (O2) for 2 h daily for three consecutive days. For control experiments, 30% O2 was delivered at the same flow rate. For drug treatment, DFP (75 mg/kg, intraperitoneally, synthesized in China Peptides Co., Ltd., Shanghai, China) or DMT1i (50 mg/kg, orally, MedChemExpress, China) was administered to the animals 1 h before GA daily for three consecutive days.

Whole cerebral neocortices were removed from the rat fetuses, and incubated in 0.03% trypsin in Earles balanced salt solution (EBSS) without Ca2 + or Mg2 + for 90 min at 37. Subsequently, the cortices washed with fresh EBSS, transferred into culture medium (12.5% F-12 growth media (Sigma), 12.5% heat inactivated horse serum (Hyclone), 30 U/ml Penicillin, 0.03 mg/ml Streptomycin solution (both Sigma), 2.5 mM glutamine (Sigma), 10 mM HEPES (Omega) in DMEM with high glucose (4500 g/l, Sigma) and dissociated by trituration.

Whole cerebral neocortices were removed from the rat fetuses, and incubated in 0.03% trypsin in Earles balanced salt solution (EBSS) without Ca2 + or Mg2 + for 90 min at 37. Subsequently, the cortices washed with fresh EBSS, transferred into culture medium (12.5% F-12 growth media (Sigma), 12.5% heat inactivated horse serum (Hyclone), 30 U/ml Penicillin, 0.03 mg/ml Streptomycin solution (both Sigma), 2.5 mM glutamine (Sigma), 10 mM HEPES (Omega) in DMEM with high glucose (4500 g/l, Sigma) and dissociated by trituration.