The developmental potential of progenitors at two final stages of the macroglial lineage giving rise to oligodendrocytes in postnatal rat brain was studied in response to defined and serum inducers of astrocyte gene expression. Cell immunoselection [with Gd3 ganglioside, O4 and galactocerebroside (GalC) antibodies] was used to isolate G+D3O4- and O4+GalC- phenotypes directly from premyelinating cerebrum. In a basal defined culture medium, G+D3O4- progenitors differentiated infrequently into oligodendrocytes on a growth substratum comprised of meningeal cell-derived extracellular matrix. Their conversion into astrocytes, as determined by immunofluorescence analysis of glial fibrillary acidic protein expression, was induced by oncostatin-M as well as leukemia inhibitory factor (LIF) and ciliary neurotrophic factor, but not interleukin-6, and required extracellular matrix. By comparison, O4+GalC- progenitors were refractory to astrocyte induction under these conditions, as in short-term cultures of optic nerve, and differentiated into myelinogenic oligodendrocytes instead. Only in response to an overriding stimulus in fetal bovine serum did O4+GalC- progenitors, like their immediate precursors, become astrocytic. These data functionally distinguish two classes of astrocyte-inducing agents to provide clear evidence of an oligodendroblast, a progenitor defined by surface phenotype (O4+GalC-) and an altered response of the oligodendrocyte lineage to cytokines using signal transducer LIFR beta.