Induction of the N-methyl-D-aspartate receptor subunit 1 in the immortalized neuronal progenitor cell line HC2S2 during differentiation into neurons.

Abstract

Conditionally immortalized neuronal progenitor cell line HC2S2 differentiates into mature neurons after suppression of the v-myc expression with tetracycline. Reverse transcription-polymerase chain reaction analyses were used to measure expression levels of N-metyl-D-aspartate receptor subunit 1 (NMDAR1) mRNAs encoding splice variants (NMDAR1a, -exon 5; NMDAR1b, +exon 5) in HC2S2 cells during the differentiation. Differential induction of NMDAR1a and NMDAR1b mRNAs was observed during the differentiation. Very low expression of NMDAR1 was observed in undifferentiated HC2S2 cells. NMDAR1a mRNA was induced coincidentally with the emergence of neurites, whereas NMDAR1b mRNA was induced at the time of network formation. Immunohistochemistry also demonstrated induction of NMDAR1 immunoreactivity in differentiated HC2S2 cells. In addition, expression of NMDAR2 mRNA and immunoreactivity was observed in undifferentiated and differentiated HC2S2 cells, suggesting that functional NMDA receptors are present in differentiated HC2S2 cells. While exposure to NMDA resulted in almost no cell death in undifferentiated HC2S2 cells, NMDA induced cell death in differentiated HC2S2 cells in a dose-dependent fashion. These findings suggest that the expression of NMDAR1 mRNA may be regulated by myc or its counterpart during neuronal terminal differentiation and that the splicing choice between NMDAR1a and NMDAR1b may vary according to the formation of neuronal network.