The development of functional gap-junctional communication was studied in cells of the oligodendrocyte lineage. The presence of cell-cell coupling was determined by the passage of current between cells using a double whole-cell patch-clamp system or by injecting the low molecular weight dye Lucifer yellow into individual cells via a patch pepette and observing the diffusion of the dye into adjacent cells. The developmental stage of the cells under study was determined using antibodies to specific surface markers (04, 01, and 010) that characterize cells of sequential maturity along the oligodendrocyte lineage (Kuhlmann-Krieg et al., 1988; Sommer and Schachner, 1981; 1982). Both stages of precursor cells of this lineage, O4+ and O4-, almost never showed dye or electrical coupling, even though they were in close physical contact with other cells. The O1-positive oligodendrocytes with simple morphology were also noncoupled, with only few exceptions. In contrast, more than 40 percent of more mature, O10-positive oligodendrocytes showed cell-cell coupling detectable with both dye and current injection. Thus, the formation of gap junctions between cells of the oligodendrocyte lineage does not occur with, but some time after, the commitment of the cell to becoming an oligodendrocyte.