INFLUENCE OF PARTIALLY C-TERMINAL DELETION OF NR2A SUBUNITS ON THE SURFACE EXPRESSION AND FUNCTION OF NMDA RECEPTOR CHANNELS / 解剖学报

ABSTRACT

Objective To investigate whether the partially C-terminal deletion of NR2A subunit alters the surface expression and channel function of NMDA receptors in both HEK293 cells and cultured hippocampal neurons of rats. Methods Four plasmids for NR2A mutants with N-terminally GFP-tagged and C-terminal deletion NR2A?C1(?897L-1017S),NR2A?C2(?1024D-1142P),NR2A?C3(?1149D-1347G),and NR2A?C4(?1354S-1464V) were generated,and transfected into HEK293 cells and hippocampal neurons in culture.Surface staining was performed using anti-GFP antibody and Cy3 conjugated secondary antibody.Glutamate evoked currents were also detected using whole-cell recording. Results Positive surface staining was found for all the HEK293 cell co-transfected NR1-1a/NR2A?C1,NR1-1a/NR2A?C2,NR1-1a/NR2A?C3 or NR1-1a/NR2A?C4,and quantitative analysis showed no significant decrease in surface expression level when compared to that from NR1-1a/NR2A transfection.Glutamate-evoked currents were recorded in HEK293 cells co-transfected with NR1-1a/NR2A?C2 or NR1-1a/NR2A?C4.Surface expression level of NMDA receptor clusters on dendrites was significantly decreased in the neurons transfected with NR2A?C1,NR2A?C2 or NR2A?C3 than in those transfected with NR2A.Conclusion C-terminal deletion of NR2A subunit differentially effects surface expression of NMDA receptors in HEK293 cells and in hippocampal neurons in culture.