Vasopressin and oxytocin neuroendocrine cells within the supraoptic nucleus of the adult hypothalamus (SON) display mRNA expression for the NMDA receptor subunits, NR1 and NR2B, NR2C and NR2D. The NR2B subunit confers slow decay kinetics (relative to NR1/NR2A receptors) and high magnesium sensitivity to NMDA receptor responses--properties which may contribute to the NMDA receptor-mediated bursting manifested by these cells. Therefore, we examined NR2B protein expression and its developmental profile in the SON and compared it to that in the cortex and cerebellum--areas which have been studied previously. We performed Western blot analysis on SON homogenates from embryonic, postnatal (PN7, 14, 21), and adult rats using an NR2B-specific antibody. Adult NR2B levels in the SON and PVN were similar but low relative to those of cortex. SON NR2B protein levels rose in the first postnatal week, remained high through PN21, and later declined to significantly lower levels in the adult. A similar profile was observed in cerebellum, where NR2B expression displayed a sharp peak at PN14 and later declined to minimal or undetectable levels in the adult. In contrast, NR2B continued to be overexpressed through adulthood in the cortex. The ontogenic pattern for NR1 expression, which included unregulation during early postnatal life and adulthood, was similar in the SON and cortex. A different pattern was observed for the cerebellum, where NR1 levels increased gradually after ED17 to reach significantly greater adult levels. Of all three areas studied, the SON displayed the earliest developmental rise in NR1 levels. SON explant cultures proved to be a useful preparation, since they contained neurons which synthesized NR1 and NR2B subunits in quantities similar to those of ED17 SON. Our findings suggest that NMDA receptors on SON neuroendocrine cells are assembled using NR1 and NR2B subunits, and that their plastic expression in early postnatal life may play a role during development.