Time-dependent changes in NMDA receptor expression in neurones cultured from rat brain

Several studies have shown marked changes in the regional expression pattern of N-methyl-D-aspartate (NMDA) receptor subunit composition in vivo in the developing brain. Similar developmental changes may also occur in vitro. Thus, displacement of [3H]MK-801 binding by the subunit-selective NMDA antagonist, ifenprodil, in membrane homogenates prepared from cultured neurones, has been shown to comprise two components, the relative proportion of which changed with time in culture [11]. In the present experiments we have used electrophysiological methods to determine the influence of time in culture on NMDA receptor subtype expression pattern in individual neurones. Shortly after plating (13 days in vitro (DIV)), approximately 70% of total NMDA-induced inward current in voltage-clamped rat cortical neurones is antagonised by ifenprodil with relatively high affinity (approximately 1 microM). By 65 DIV, however, the high-affinity component contributed to only approximately 20% of the overall antagonism. Cerebellar granule cells also appear to undergo a similar change in their NMDA receptor expression with the exception that, in general, they appear initially to show considerably less of the ifenprodil high-affinity component and this disappears completely by 15 DIV. These experiments suggest that individual cortical and granule cells express at least two different NMDA receptor subtypes and that their relative proportion changes with time in culture.