An antisense construct reduces XXXX receptor 2A expression and receptor-mediated excitotoxicity as determined by a novel flow cytometric approach.

@entity11588 @entity1024 @entity63 @entity6806 @entity179 @entity1566
The @entity1024 receptor (NMDAR) is a major neurotransmitter receptor in the central nervous system (CNS), with functional roles in learning, memory, and sensation. Several mechanisms potentiate NMDARs, and NMDAR hyperexcitability plays pathophysiological roles in many conditions, such as @entity179 , @entity63 , and chronic conditions arising from @entity1566 . Previous research suggests that the @entity11588 subunit of the receptor contributes to NMDAR excitotoxicity in heterologous cells and in neurons in vivo. To investigate the role of @entity11588 in NMDAR excitotoxicity, we have developed a system based on flow cytometry that allows rapid evaluation of the effect of antisense constructs on protein expression and channel function. The enhanced yellow fluorescent protein (EYFP) was fused to obligatory NMDAR subunits, allowing expression to be monitored in living cultured cells. An @entity11588 antisense construct, asNR2A, specifically and effectively reduced @entity11588 -EYFP expression. @entity6806 and @entity11588 fusion proteins formed functional, excitotoxic channels upon co-expression. The asNR2A RNA significantly reduced NMDAR excitotoxicity when @entity11588 levels were limiting for channel formation. Using our assay system, further optimization can be achieved rapidly. The asNR2A construct and the assays developed for this study can be used to provide insights into NMDAR biology and disease.
