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The Journal of Neuroscience, October 15, 2002, 22(20):8902-8910
Relationship between Availability of NMDA Receptor Subunits and Their Expression at the Synapse
Kate Prybylowski1, Zhanyan Fu2, Gabriele Losi2, Lynda M. Hawkins1, JianHong Luo2, Kai Chang1, Robert J. Wenthold1, and Stefano Vicini2 1 Laboratory of Neurochemistry, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, Maryland 20892, and 2 Department of Physiology and Biophysics, Georgetown University Medical Center, Washington, DC 20057
The effect of increasing the expression of NMDA subunits in cerebellar granule cells (CGCs) by transfection was studied to determine how the availability of various NMDA subunits controls both the total pool of functional receptors and the synaptic pool. Overexpression of either NR2A or NR2B, but not splice variants of NR1, by transfection caused a significant increase in the total number of functional NMDA receptors and in surface NR1 subunit cluster density in CGCs in primary culture. These data solidify the central role of NR2 subunit availability in determining the number of cell surface receptors. Overexpression of either NR2A or NR2B significantly altered the deactivation kinetics of NMDA-mediated miniature EPSCs (NMDA-mEPSCs). However, there was no significant effect of NR2 subunit overexpression on the mEPSC amplitude or single-channel conductance. NR2 subunit overexpression did not change the rate of block by MK-801 of NMDA-mediated currents in excised patches from CGCs, indicating that subunit composition does not regulate peak open probability of the channel in CGCs. With the overexpression of a mutant of NR2B lacking the PDZ binding domain, there was an increase in the total number of NMDA receptors without a change in mEPSC kinetics. Therefore, the entry of NMDA receptors into the synapse requires a PDZ binding domain and is limited by means other than receptor subunit availability.
Key words: cerebellar granule cells; ifenprodil sensitivity; NMDA-mEPSCs; peak open probability; PDZ binding domain; receptor targeting
Copyright © 2002 Society for Neuroscience 0270-6474/02/22208902-09$05.00/0
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