NMDA受体在爪蟾卵母细胞和HEK-293细胞表达体系中的建立

摘要

Objective:To establish Xenopus laevis oocytes and HEK-293 cells models expressing N-methyl-D-aspartate receptors (NMDAR) comprising NR2AR/NR2AR subunits.Methods:Microinjection and liposome transfection were performed respectively to establish Xenopus laevis oocytes and HEK-293 cells expressing NMDAR.Currents induced by NMDAR subunits NR2AR and NR2BR were recorded by whole-cell patch-clamp.Results:NMDAR subtypes NR2AR and NR2BR-mediated whole cell currents were detected.The success rate of NMDAR gene injection was 100％ in Xenopus laevis oocytes,whereas NMDAR mediated currents were not detected in the HEK-293 cells,and the transfection success rate was (6.7±1.39)％.Conclusion:NMDAR was expressed in Xenopus laevis oocytes,providing a cell model to investigate the effects of drugs on the current of NMDAR.However,we failed to examine the currents in HEK-293 expression system,which might be related to low transfection efficiency.%目的:建立特异性表达NMDA受体(NMDAR) NR2AR、NR2BR亚型的爪蟾卵母细胞和HEK-293细胞模型,选出合适的表达体系模型,为研究药物对NMDAR介导电流的影响奠定基础.方法:分别采用显微注射法和脂质体转染法将NMDAR表达在爪蟾卵母细胞和HEK-293细胞上,利用全细胞膜片钳技术,记录NMDAR亚型NR2AR、NR2BR介导的电流.结果:在建立的爪蟾卵母表达体系中,NMDAR基因注射成功率为100％,且能检测到NMDAR亚型NR2AR和NR2BR介导的内向电流,而在HEK 293表达体系中,转染成功率为(6.7±1.39)％,未检测到NMDAR介导的电流.结论:成功建立爪蟾卵母细胞NMDAR表达体系,可作为研究药物对爪蟾卵母细胞表达体系上NMDAR电流影响的合适的细胞表达体系模型.在HEK-293表达体系上未检测到相关电流,可能与其转染后细胞活性大大降低、脂质体转染效率低等因素有关.