NMDAR1单克隆抗体抑制谷氨酸诱导的大鼠海马神经元Ca2+内流

目的观察人N-甲基-D-门冬氨酸受体(NMDAR,NR)主亚基(NR1)单克隆抗体mAbN1对谷氨酸诱导的大鼠海马神经元Ca2 内流的影响。方法建立谷氨酸介导的大鼠海马神经元兴奋毒性损伤模型,以mAbN1及MK-801分别预处理海马神经元,用Fluo-3/AM法,在激光扫描共聚焦显微镜下观察对细胞内游离Ca2 浓度([Ca2 ]i)的影响。结果mAbN1能显著抑制谷氨酸所致海马神经元[Ca2 ]i升高,此作用强于MK-801,且其本身对生理状态下神经元[Ca2 ]i无影响。结论mAbN1的抗兴奋毒性作用可能是通过改变NR的蛋白质二级结构从而影响兴奋毒性作用中的Ca2 内流实现的。

Anti-NR1 monoclonal antibody inhibits the glutamate-induced intracellular free calcium increase in rat hippocampal neurons

Objective To study the effect of mAbN1,a monoclonal antibody against N-methyl-D-aspartate receptor subunit 1(NR1)on Ca2 influx after glutamate stimulation in cultured rat hippocampal neurons.Methods Excitotoxicity was induced by glutamate in cultured hippocampal neurons.Confocal laser scanning microscopy was used to observe the changes in intracellular free calcium concentration([Ca2 ]i)at the level of cultured hippocampal neurons following pretreatment with mAbN1 and MK-801.Intracellular free calcium was imaged after loading cells with the fluorescent dye indicator fluo-3/AM.Results Our findings indicate that as compared with MK-801,mAbN1 can more significantly attenuate the glutamate-induced [Ca2 ]i increase,and it has no effect on [Ca2 ]i in physiological condition.Conclusion mAbN1 may alter the secondary structure of NR,consequently influence Ca2 influx in excitotoxicity process.