Researchers have developed many innovative ways to investigate the neural circuitry of hippocampus. However, it is still a difficult task because of its complexity and inaccessibility. Recently, a multi-electrode array system has been adapted to deal with the hippocampal neural circuit. With this tool, we have studied the spatiotemporal synaptic transmission changes in hippocampal slice culture. Three to five weeks old Sprague-Dawley mail rats were humanely killed by instant decapitation using a Guillotine and then whole brain was removed. Acute hippocampal slices were obtained by conventional methods. The experiments were performed in accordance with the animal care guidelines of the Korean Academy of Medical Sciences and planned to minimize the number of animals and their suffering. MEA system (multielectrode array; MCS GmbH, Germany) were used in the study. Train pulse (140μs biphasic, 3Hz for 4min and 65μA current) was applied to a selected channel at every hour for 6 hours (STG1004; MCS GmbH) and the responses were recorded in other 59 channels using Recorder-Rack software (MCS GmbH). Analysis of data was carried out by a histogram and pseudocolor imaging analysis methods that were developed in our laboratory. Ensemble of acute hippocampal slice and the MEA system has been successfully adapted to reveal the high-resolution spatiotemporal synaptic transmission changes in hippocampal brain slice.