Astroglia inhibit the proliferation of neocortical cells and prevent the generation of small GABAergic neurons in vitro.

We quantitatively studied the dynamics of rat neocortical precursor proliferation in vitro, and additionally examined the effects of neuron-glia interactions on the proliferation and differentiation of neurons, and particularly of gamma-aminobutyric acid (GABA)-containing cells. In cultures grown on glia-free substrate, cellular proliferation was detected at least until the end of the second week in vitro, but most neurons which expressed detectable amounts of microtubule-associated protein at 12 days in vitro were generated early during the first week. Further double-labelling experiments, combining 5'-bromo-2'-deoxyuridine with GABA or beta-tubulin III immunohistochemistry, provided direct evidence that neuronal proliferation continued through the second week in vitro, and that a population of small GABAergic neurons was generated between 3 and 12 days in vitro. Culturing cells on a glial substrate significantly reduced the generation of small GABAergic cells and strongly inhibited the total cell proliferation. Inhibition also occurred if astrocytes were added to the culture after 6 days in vitro, but was significantly decreased if cells were grown on a fixed glial substrate, suggesting that the effect might be at least partially mediated by active interactions between neurons and glia. In conclusion, our results show that the sustained proliferation of precursor cells in neocortical cultures is necessary for the differentiation of small GABAergic neurons, and that mature astroglia effectively inhibit the proliferation of neocortical precursors thereby affecting the appearance of a population of GABAergic cells.