Channel properties of NMDA receptors on magnocellular neuroendocrine cells cultured from the rat supraoptic nucleus

Application of N-methyl-d-aspartate (NMDA) to the supraoptic nucleus of the hypothalamus (SON) generates clustered firing that may be important in hormone release. However, synaptically evoked EPSPs recorded from SON neurons exhibit varying contributions from NMDA receptors. We used the high resolution of single-channel recording to examine the receptor and ion channel properties of NMDA receptors expressed by SON neurons in `punch' culture. Biocytin introduced into individual neurons during patch clamp recording revealed large (32.1±3.3 μm), oblong somas and bipolar extensions typical of magnocellular neuroendocrine cells (MNCs). Rapid application of NMDA (100–300 μM) in the presence of 10 μM glycine to outside-out macropatches resulted in openings with an average conductance of 46.9 pS and reversal potential of +3.9 mV. Increasing glycine from 0.03 to 30 μM increased the apparent frequency, duration and occurrence of overlapping NMDA-elicited openings. NMDA responses were inhibited by Mg²⁺ in a voltage-dependent manner and by the NMDA-site antagonist, d-(−)-2-amino-5-phosphonovaleric acid (D-APV). Application of saturating NMDA or glycine alone with the glycine-site antagonist, 5,7-dichlorokynurenate (DCK) or with D-APV, respectively, did not result in agonist-induced openings. NR1 immunoreactivity was observed in large neurons (>25 μm) with MNC-like morphology. These single-channel and immunocytochemical data confirm the presence of functional NR1-containing NMDA receptors in MNCs.