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目的观察原代培养神经元在红藻氨酸(KA)作用下细胞表面形态的三维构像变化。方法利用原子力显微镜(AFM)对大鼠海马神经元经不同浓度的KA(25和250 μmol/L,50和100 nmol/L)分别作用10 min和100 min后的表面结构进行纳米级水平扫描和观测。结果正常海马神经元表面光滑,起伏均匀、规律;KA作用后神经元呈退行性改变, 表现为胞体肿胀,胞膜表面粗糙,出现隆起和“孔洞”样结构,并且其变化程度分别与作用时间和KA浓度呈量-效关系。结论KA对海马神经元毒性作用后胞膜表面超微结构所产生的明显变化,可借助AFM清晰观察,并定量分析。
Objective To observe the three-dimensional conformational changes of the cell surface morphology of primary cultured neurons under the action of kainic acid (KA). Methods Atomic force microscopy (AFM) was used to scan the surface structures of rat hippocampal neurons after exposure to KA (25 and 250 μmol / L, 50 and 100 nmol / L) for 10 min and 100 min, respectively. Observation. Results The neurons in normal hippocampus had a smooth, uniform surface and regular pattern. After KA, the neurons showed degenerative changes, showing swelling of cell body, rough surface of cell membrane and uplift and “hole” -like structures. The changes were related to the action time And KA concentration in a dose-effect relationship. Conclusion The obvious changes of membrane surface ultrastructure induced by KA on the neurons of hippocampus can be clearly observed and quantitatively analyzed by AFM.