A new method allowing long-term potentiation recordings in hippocampal organotypic slices.

BACKGROUND

Hippocampal organotypic slices are used to improve the understanding of synaptic plasticity mechanisms because they allow longer term studies compared to acute slices. However, it is more delicate to keep cultures alive in the recording system outside  conditions. Experiments from the organotypic cultures are common but the handling of slices is rarely described in the literature, even though tissue preservation is crucial. Instruments are sometimes required to extract the slices from the culture inserts but this approach is delicate and can lead to damage, given how strongly the slices are attached to the insert.

METHODS

A new configuration is proposed to secure the transfer of slices from the incubator to the recording chamber through an adaptor piece that can be designed for any model of chamber and/or insert. The adaptor is a Plexiglas ring in which a culture insert containing the slice can be easily introduced and stabilized. This system allows slices to be placed in the interface for electrophysiological investigations without having to detach them from the insert. That way, no damage is caused and the recording system can safely hold the slices, maintaining them close to culture conditions.

RESULTS

In addition to the description of the adaptation system, slices were characterized. Their viability was validated and microglial expression was observed. According to the experimental conditions, neuroprotective ramified microgliocytes are present. Dendritic spines studies were also performed to determine neuronal network maturity in culture. Moreover, SKF 83822 hydrobromide and three trains of 100 pulses at 100 Hz with a 10-min inter-train interval are suggested to induce long-term potentiation and to record an increase of fEPSP amplitude and slope.

CONCLUSION

This paper provides detailed information on the preparation and characterization of hippocampal organotypic slices, a new recording configuration more suitable for cultures, and a long-term potentiation protocol combining SKF and trains.