目的了解大鼠耳蜗螺旋神经节神经元(SGN)中NMDA受体亚单位的基因表达水平,为进一步探讨SGN中NMDA受体的功能提供依据。方法利用倍比稀释的出生3 d的大鼠正常脑组织CDNA构建NMDA受体亚单位NR1、NR2(A~D),NR3(A~B)与甘油醛-3磷-酸脱氢酶(GAP-DH)基因的相对标准曲线,通过实验验证NR1、NR2(A~D),NR3(A~B)分别和内参基因GAPDH是否有一致的扩增效率,应用△Ct法对NMDA受体各亚单位的表达进行相对定量。结果 NMDA受体亚单位中由高到低依次为NR2B、NR3A、NR1、NR2D、NR3B、R2A、NR2C;除NR2D与NR3B外,其余各亚单位之间比较均有统计学意义(P〈0.01)。结论实时荧光定量PCR可以对SGN中NMDA受体亚单位的表达进行全面定量分析,进一步证实了NMDA受体各亚单位在耳蜗SGN中的表达水平。
Objective To detect the expressions of the rat spiral ganglion neurons(SGN) NMDA receptor subunit gene in order to provide the basis for the futher study of NMDA receptor function.Methods The realtive standard curve of the rat NMDA receptor subunits NR1,NR2A-NR2D,NR3A-3B and glyceraldehyde-3-phosphate dehydrogenase(GAPDH) gene established by dilution the cDNA got from normal brain tissue of rat,The amplification efficiencies of the target(NR1,NR2A-NR2D,NR3A-3B) were tested and the reference(GAPDH)genes tested for the △Ct calculaion to be valid.Results The rat NMDA receptor subunits NR1 had the highest expression followed by NR3A,NR1,NR2D,NR3B,NR2A,NR2C.The difference was statistically significant(P0.01).Conclusion Real-time PCR can give a comprehensive quantitative analysis for the SGN NMDA recepter subnits,and futher confirm the expression of the NMDA receptor subnit in the SGN.