Spontaneous neuronal discharge patterns in developing organotypic mega-co-cultures of neonatal rat cerebral cortex

Sagittal slices of neonatal rat neocortex, extending from the prefrontal to the occipital area, were cultured separately or in pairs, oriented in such a way that axons projecting from the ventricular surface of each explant could innervate the other one. Functional connections were made between as well as within the explants, and spontaneous field potentials and associated action potentials occurred in variable bursts, and with varying degrees of synchrony. Spike-train analysis revealed that the activity patterns seen in these ‘mega’ co-cultures closely mimic ‘tracé alternant’ patterns, consisting of trains of burst discharges recurring several times per minute, which are characteristic for the immature intact cerebral cortex during slow-wave sleep. The prefrontal region was consistently less active than the occipital area but the two were qualitatively similar with respect to their patterns of neuronal firing. Isolated mega-cultures, on the other hand, despite their large size, exhibited only intermittent brief bursts that closely resembled those observed both in occipital cortex tissue fragments and in dissociated cell cultures. The mega-co-culture preparation thus appears to give the best currently available approximation to intrinsic cerebral discharge patterns in vivo.