Culturing hippocampal neurons.

The procedure for making a low density culture of hippocampal neurons has been elaborated by Goslin and Banker. The viability of hippocampal neurons, which are sparsely disseminated on the glass surface, is maintained by a separately cultured glial monolayer; the glial feeder layer is grown on the bottom surface of the dish, while those neurons, placed face down, are attached on the coverslips. This method is originaLly designed for the observation of the maturation, polarity and axogenesis of a single neuron. In addition, this method can be applied for a variety of other purposes: (1) to observe synaptogenesis, (2) to analyze synaptic function electrophysiologically, (3) to analyze receptor functions and signaling cascades pharmacologically, (4) to visualize a molecular dynamics by time-lapse analyses of GFP-tagged molecules, and (5) to observe ultrastructure by an electron microscope. Furthermore, these neurons are useful even in biochemical experiments because they are relatively uniform without glial contamination and highly enriched in synaptic components.