Overexpression of glyceraldehyde-3-phospahe dehydrogenase is not involved in 5-hydroxytryptamine (5-HT)-induced necrosis in cultured cerebrocortical neurons.

Cerebrocortical cell cultures were prepared from 1-d-old rats. On post-culture day 6, 5-hydroxytryptamine (5-HT) was added to the medium and cells were exposed for another 3 d. 5-HT elicited cytotoxicity in a dose-dependent manner, and the survival rate of neuronal cells was decreased to 64.9+/-5.0% at 0.1 mM concentration. Chromatin staining with Hoechst 33258 and electron microscopy revealed that the 5-HT-induced neuronal death was entirely due to necrosis. Pretreatments with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antisense oligonucleotide and several classical apoptotic inhibitors did not exhibit neuroprotection in this paradigm. Northern blot analysis showed that the enhancement of GAPDH mRNA levels was undetected during cell death. The present results demonstrate that GAPDH overexpression is not involved in the 5-HT-induced necrotic death pathway.