Cognitive mapping functions of the hippocampus critically depend on the recurrent network of the CA3 pyramidal cells. However, it is still not known in detail how network activity patterns emerge, or how they encode information. By using functional multineuron calcium imaging, we simultaneously recorded the activity of >100 neurons in the CA3 region of hippocampal slice cultures. We utilized a novel computational method to analyze the multichannel spike trains and to depict functional neuronal assemblies. By means of event synchronization and the correlation matrix analysis method, we found that: 1), the average functional neuronal cluster consists of 23 neurons, and neurons could be part of multiple assemblies; 2), the clustering strength, size, and mean distance among cells in neuronal assemblies follow a power-law-like distribution; 3), the clustering strength and size of neuronal assemblies are not correlated with the total number of neurons and their physical distance; and 4), the clustering distance of neuronal assemblies is weakly correlated with the total number of neurons and their physical distance. These findings suggest that the functional organization of the spontaneously firing CA3 hippocampal network is a scale-free structure in slice culture.
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