In this study we have performed a detailed analysis of EGF-responsive precursors as they develop into neurons and astrocytes using antibodies to nestin, microtubule-associated protein 2 (MAP-2c and MAP-2ab) and glial fibriallary acidic protein (GFAP). Surprisingly, at early time points, most GFAP-positive cells also stained for MAP-2c, and we postulate that this may be a normal stage of astroglial development. At 7 days most of the cells had developed into astrocytes and MAP-2ab-positive cells only represented 5% of the total neuronal population. This study shows that (i) MAP-2c is a marker for early precursors, (ii) the majority of cells developing from. EGF-responsive precursors develop into glia and (iii) only a small population of cells arising from expanded populations of EGF-responsive precursors develop into neurons expressing MAP-2ab. Thus, certain critical signals important for full neuronal differentiation may be missing from this system.