| Objective To study the NMDA receptor subunit and Postsynastic density 95 (PSD-95) mRNA expression following Mg2+-free-induced seizures in cultured developing neuron. and explore the effect of early seizure-like discharge on MTT metabolic rate in developing cortical neurons of rat.Methods The seizure model was recurrent epileptiform discharges induced by transient magnesium-free media treatment in primary cultured embryo cortical neurons of rats. According to different treatment to cortical neurons at cultured day 6.,three groups were divided:regular Neurobasal/B27 medium(CONT),regular extracellular solution medium(PS),and magnesium-free medium(MGF).The neurons were exposed to above three media for 3h in culture day 6,and then cultured in regular Neurobasal/B27 medium again.. The expression of NMDAR subunit of NR1,NR2B mRNA and PSD-95 mRNA were measured using fluorescent real-time quantitative PCR and MTT metabolic rate (%) at 6h,24h,72h and day 6 (in cultured D12) after treatment.Results Compared with group CONT and PS respectively, in group MGF the expression of NR1 mRNA decreased at 24h(p<0.05) and increased at day 6(p<0.05); the expression of NR2B mRNA increased at 6h (p<0.05);the expression of PSD-95 increased at 24h (p<0.05). Compared with group CONT and PS respectively, in group MGF the MTT metabolic rate (%) at 6h,24h,72h and day 6 after treatment. were 65.32%,78.9%,,66.74%,72.56%,there were no statistic significant difference of MTT metabolic rate in PS and CONT.Conclusion The change of NMDAR subunit mRNA expression were resulted not only by short-term changes of early magnesium-free medium but also long-term changes in developing cortical neurons of rat. NR2B and PSD-95 may play a important role in the early seizures. The effect on the mitochondrial function of developing neurons after magnesium-free-induced seizure-like discharge may from the acute phase which lasted for a long period of time.
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