Oligodendrocytes elaborate an extensive membrane network that ensheathes CNS axons in multilamellar wrappings. A compaction process excludes much of the cytoplasm in mature myelin membranes, giving rise to distinct lipid/protein compositions in two membrane compartments (compact myelin and membranes of the cell body and processes). Insofar as oligodendrocytes arise from neuroepithelial progenitors, it seems likely that some elements are shared for protein targeting by these two cell types. We hypothesized that certain membrane proteins targeting different oligodendroglial membrane compartments would preferentially sort to opposite domains when transfected into Madin-Darby canine kidney (MDCK) epithelial cells. Myelin/oligodendrocyte glycoprotein (MOG) is found in uncompacted membrane (cell body, processes), and it sorts exclusively to MDCK basolateral membrane. Proteolipid protein (PLP) is found in compact myelin, and it sorts exclusively to MDCK apical membrane. Myelin-associated glycoprotein (MAG) is primarily in the periaxonal inner loop of myelin; however, it fails to target preferentially within MDCK cells. This inability of MAG to sort within MDCK cells suggests a lack of required oligodendroglial-specific targeting components. In contrast, the sorting machinery in both oligodendrocytes and MDCK cells recognizes targeting signals for MOG and PLP, and we propose that these oligodendroglial membrane proteins delineate cognate basolateral and apical domains, respectively. J Neurosci. Res. 66:1140–1148, 2001. © 2001 Wiley-Liss, Inc.