To examine the role of extracellular matrix (ECM)/integrin interactions in myelination we have analyzed oligodendrocyte differentiation and myelin membrane formation in oligodendrocytes grown in cell culture. We have found that the ECM substrates fibronectin, vitronectin, and laminin-2 (merosin) have no effect on differentiation, as measured by the appearance of myelin basic protein-expressing cells, but that laminin-2 substrates dramatically enhance myelin membrane formation. Blocking antibody and immunolocalization studies suggest that this effect is mediated via β1 integrins. The αv integrins expressed on oligodendrocytes, in contrast, are less effective at promoting membrane formation. These results show that the interaction between laminin-2 expressed in white matter tracts and oligodendrocyte laminin- binding integrins may be an important part of the signalling mechanisms that stimulate oligodendrocytes to elaborate the extensive myelin membrane required to wrap the axon and form the myelin sheath. The results also provide a logical explanation for the abnormalities of myelination observed in humans with merosin-deficient congenital muscular dystrophy.