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目的对比观察正常培养皮层神经元在N-甲基-D-天冬氨酸(NMDA)及其受体拮抗剂—MK-801作用下膜表面三维构像形态的改变。方法利用分辨率为0.1~0.01nm的原子力显微镜(AFM)对原代培养大鼠皮层神经细胞膜表面进行纳米尺度的扫描观测。结果正常神经元膜表面光滑,起伏均匀,隆起的颗粒状蛋白密集,间隔规律;NMDA损伤后神经元破碎,崩解,膜失去连续性;NMDA+MK-801作用下神经元膜皱折增加,边缘粗糙,起伏程度介于前两者之间。结论(1)AFM具有分辨率高,制样简单特点。(2)AFM能细微地分辨损伤保护作用后引起的细胞膜表面三维形态改变。(3)NMDA作用后膜结构开始解体,膜蛋白颗粒聚集增大,脂质凹陷加深,间距增宽,表面粗糙度增加。
Objective To compare the morphological changes of three-dimensional conformation of membrane surface in normal cultured cortical neurons under the action of N-methyl-D-aspartate (NMDA) and its receptor antagonist-MK-801. Methods The surface of primary cultured rat cortical neurons was scanned by nanoscale using atomic force microscopy (AFM) with a resolution of 0.1 ~ 0.01 nm. Results The normal neurons showed smooth and uniform surface, with densely populated granular proteins and regular intervals. The neurons were broken, disintegrated and the membrane lost continuity after NMDA injury. The neuronal membrane wrinkles increased under the action of NMDA + MK-801, Rough edges, undulation between the first two. Conclusion (1) AFM has high resolution, simple sample preparation. (2) AFM can finely distinguish the three-dimensional morphological changes of cell membrane surface caused by injury protection. (3) After the action of NMDA, the membrane structure began to disintegrate, the aggregation of membrane protein particles increased, the lipid depressions deepened, the spacing broadened and the surface roughness increased.