Culturing hippocampal neurons
Culturing hippocampal neurons
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DOI：
10.1038/nprot.2006.356
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Banker, Gary
中科院分区：
文献类型：
--
作者：
Kaech, Stefanie;Banker, Gary
我们提供了从胚胎大鼠或小鼠制备海马神经元低密度离散细胞培养物的实验方案。神经元培养在多聚赖氨酸处理过的盖玻片上，这些盖玻片悬浮在星形胶质细胞饲养层上方，并维持在无血清培养基中。按照该方案培养时，海马神经元会适当极化，形成广泛的轴突和树突分支，并彼此形成大量功能性突触连接。海马神经元培养物已被广泛用于观察内源性或表达的蛋白质的亚细胞定位、蛋白质运输成像以及确定神经元极性发育、树突生长和突触形成的分子机制。胶质饲养层培养物的制备必须提前2周开始，准备用作神经元生长底物的盖玻片需要5天。解剖海马并铺板海马神经元需要2 - 3小时。
We provide protocols for preparing low-density dissociated-cell cultures of hippocampal neurons from embryonic rats or mice. The neurons are cultured on polylysine-treated coverslips, which are suspended above an astrocyte feeder layer and maintained in serum-free medium. When cultured according to this protocol, hippocampal neurons become appropriately polarized, develop extensive axonal and dendritic arbors and form numerous, functional synaptic connections with one another. Hippocampal cultures have been used widely for visualizing the subcellular localization of endogenous or expressed proteins, for imaging protein trafficking and for defining the molecular mechanisms underlying the development of neuronal polarity, dendritic growth and synapse formation. Preparation of glial feeder cultures must begin 2 weeks in advance, and it takes 5 d to prepare coverslips as a substrate for neuronal growth. Dissecting the hippocampus and plating hippocampal neurons takes 2-3 h.