At embryonic stages of development, oligodendrocyte precursors (OPCs) generated in the preoptic area colonize the entire optic nerve (ON). Different factors controlling migration of ON OPCs have been identified, including secreted growth factors, morphogens and guidance cues, as well as cell adhesion molecules. We have shown previously that the soluble form of the extracellular matrix (ECM) protein anosmin-1, impairs OPC migration induced by FGF-2. In the present work, we show that anosmin-1 is expressed by both migrating OPCs and axons of the retinal ganglion cells in the embryonic ON. In vitro, we observe that OPC migration is strongly impaired by contact with anosmin-1 when used as a substrate and, in contrast to previous results, this effect is independent of FGF-2/FGFR1 signaling. We also show that OPCs preferentially adhere to anosmin-1 when compared with other ECM molecules used as substrates, and that when the endogenous anosmin-1 expressed by OPCs is blocked, OPC adhesion to all the different substrates (including anosmin-1), is significantly reduced. This novel effect of anosmin-1 on cell adhesion is also independent of FGF-2/FGFR1. We finally demonstrate that the blockade of the endogenous anosmin-1 expressed by OPCs impairs their migration. Our data suggest that the endogenous anosmin-1 expressed by OPCs is necessary for the correct adhesion of these cells to the different components of the ECM (including anosmin-1 itself), contributing to the migration of these cells.