原代培养海马神经元Drebrin和Synaptophysin的表达与动态变化

The expression and dynamic changes of drebrin and synaptophysin in primary cultured hippocampal neurons

目的:观察原代海马神经元不同时期树突棘的变化以及树突棘蛋白Drebrin和突触囊泡膜蛋白SYN的表达水平。方法:光镜观察原代培养7、14和20 d的海马神经元的形态改变。应用免疫荧光对Drebrin和SYN进行染色,观察树突棘形态和数量的改变。进一步应用Western Blot检测Drebrin和SYN蛋白的表达变化。结果:培养7 d的海马神经元,未见树突棘结构,SYN斑点状阳性产物较少。随着海马神经元培养时间的加长,突起数量逐渐增加,14 d和20 d的树突棘数量和SYN阳性产物均显著增加。Drebrin和SYN蛋白表达水平也随培养时间的加长而显著增加。结论:原代培养的海马神经元在14 d已有树突棘形成,20 d显示出成熟树突棘的形态和数量。Drebrin和SYN的蛋白表达水平反映了树突棘和突触的形成和功能状态。

Objective： To observe the changes of dendritic spines in different stages of primary cultured hippocampal neurons and the expression level of dendritic spines protein drebrin and synaptic vesicle membrane protein synaptophysin （SYN）. Methods： Morphological changes of hippocampal neurons were observed at 7 d, 14 d and 20 d. Immunofluores- cence staining of drebrin and SYN was performed to observe the changes of morphology and number of dendritic spines. The expression of drebrin and SYN were detected by Western Blot. Results： Cultured hippocampal neurons of 7 d, there were no dendritic spines, and SYN positive spots were less. With the increase of time for culturing hippocampal neuron, the number of neurites increased gradually. At 14 d and 20 d, the number of dendritic spines and SYN positive spots were significantly increased. The expression level of drebrin and SYN were significantly increased with the increase of culture time. Conclusion： The primary cultured hippocampal neurons had dendritic spines at 14 d, and the dendritic spines matured at 20 d. The expression level of drebrin and SYN reflects the formation and functional status of dendritic spines and synapses.