Ultrastructural differentiation and synaptogenesis in aggregating rotation cultures of rat cerebral cells

Cerebral cortices from fetal rats were dissociated into single cells by either trypsinization or mechanical sieving. Then the cells were allowed to form aggregates in rotation cultures. At 7, 14, 21, 28, and 35 days, aggregates were processed for electron microscopic study of morphological differentiation with special emphasis on synaptogenesis. Whether the tissue was initially dissociated by trypsin treatment or by mechanical sieving, the aggregating cultures did not exhibit any apparent differences in ultrastructural differentiation and synaptogenesis. At day 7, most neurons were immature and the extracellular space was large. Cell processes had not yet branched extensively but did contain numerous microtubules. A few immature synapses were observed. Astrocytes and oligodendrocytes displayed many of their typical cytological features. At day 14, dendritic spines had developed, some of which formed axodendritic spinous synapses. Multilayered myelin sheaths were tightly wrapped around axons. At day 21, synapses appeared mature and their number per unit area was maximal. The extracellular space had greatly decreased. At this age, asymmetrical synapses had increased approximately sixfold, whereas symmetrical synapses increased only fourfold when compared with the 7-day-old aggregates. Multifocal degeneration became apparent at 28 days and was accompanied by a significant decline in the number of synapses.