Intercellular communication mediated by gap junctions was investigated during oligodendrocyte differentiation in primary and secondary cell cultures from newborn and adult rats. Two types of communication were considered: ionic coupling and dye-coupling between similar oligodendrocytes selected at the same stage of differentiation (homotypic) and dye-coupling between oligodendrocytes and astrocytes (heterotypic). Intercellular diffusion of fluorescent probes and double whole-cell recordings were used to test the incidence of dye and ionic communication respectively. Progenitor cells, identified with A2B5 antibodies, were characterized by the absence of ionic and dye-coupling, whereas oligodendrocytes, identified with galactosylceramide antibodies, exhibited both types of communication. This homotypic coupling was inhibited by various uncoupling agents, but unaffected by treatments which increased the intracellular concentration of cAMP. In cocultures of astrocytes and oligodendrocytes, Lucifer yellow and sulphorhodamine B were exchanged in both directions. This heterotypic dye-coupling, which could be blocked by octanol, first appeared after 3 weeks in culture and increased to an incidence of 25% after 6 weeks, a developmental pattern comparable to homotypic dye-coupling between oligodendrocytes. In contrast, during the same period, progenitors and microglia were never observed to be dye-coupled with astrocytes.