Organotypic culture techniques and study of neural toxicity

Chunxi Han, Nahoko Kasai, and Keiichi Torimitsu
Materials Science Laboratory

@Organotypic slice culture technique of rat hippocampus is very important tool to study the physiological and pharmacological properties of neuron and neuronal circuits. Because the slices retain the cytoarchitecture and neuronal circuits of the tissue and because they are relatively easy to prepare, it is one of the best materials to study long-term studies of neuronal network. In epileptic animal models and human temporal lobe epilepsy, many studies have reported that the CA1 and CA3 sectors of the hippocampus are susceptible to neuronal cell death. However, little attention has been paid to neuronal cell death in CA2 sector.
@In this study, we applied the rat hippocampal slice culture technique to test whether the CA2 sector is more resistant than other sectors to chronic stimulation with bicuculline (BiC), a GABAA receptor antagonist [1][2]. The results indicated that selective neuronal cell death in the CA2 sector is induced by exposure to BiC for 12 h (Fig. 1A). After 24 h, cell death was observed in an extended area, predominantly in the CA3 sector. We investigated the type of voltage dependent calcium channel (VDCC) involved in the cell death. It was revealed that the neuronal cell death process involves a Ca2+ influx (Fig. 1B) via P/Q-type VDCC [1].

[P] C. Han, N. Kasai, K. Torimitsu, Neuroreport 16 (2005) 333-336.
[2] C. Han, N. Kasai, K. Torimitsu, Bul. JSN. 42 (2003) 238.
Fig. 1. Time- and region-dependent neuronal cell death (A) and Ca2+ influx (B) were induced by exposing the hippocampal slice cultures to BiC.