Microtubule associated protein (MAP1B) is present in cultured oligodendrocytes and co-localizes with tubulin.
Differentiation of oligodendrocytes is accompanied by the extension of processes and the assembly of the myelin membrane. It is likely that the cytoskeleton plays an important role in this process in terms of changes in cell shape, transport of myelin components, and organization of the myelin membrane. Oligodendrocytes contain microtubules (MT) which associate with other components of the cytoskeleton, and microtubule associated proteins (MAPs) may mediate some of these interactions. In this study we have shown the presence of MAP1B in oligodendrocytes grown in primary glial cultures by double-label immunofluorescence using antibodies to galactocerebroside (GC) and MAP1B. The staining of the cultures showed that GC-positive oligodendrocytes were also stained with MAP1B antibodies. However, MAP1B stain was limited to cell bodies and processes, whereas GC stain was also seen in flattened membrane sheets and punctate staining in processes. MAP1B staining was also compared with that of myelin proteolipid (PLP), myelin basic protein (MBP) and beta-tubulin in secondary glial cultures that were enriched for oligodendrocytes. The results showed a typical staining of cell bodies and membranous profiles using PLP antibodies, and the staining of cell bodies and flattened regions of membranous sheets by MBP antibodies. In contrast, both polyclonal and monoclonal antibodies to MAP1B showed a uniform diffuse staining of cell bodies, major processes, and fine interconnected processes. Double-labeling of the cells showed that MAP1B was co-localized with tubulin, but was not present in glial fibrillary acidic protein (GFAP)-positive astrocytes. Western and Northern blot analyses of primary glial cultures showed that MAP1B had a molecular mass of 320 kDa and a mRNA of 10 kb. These values are identical to those previously reported for brain MAP1B (Safaei and Fischer, 1989) and demonstrate the presence of MAP1B in oligodendrocytes.
Fischer
I
I
Department of Biochemistry, E.K. Shriver Center, Waltham, Massachusetts 02254.
Konola
J
J
Cochary
E
E
D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000276:immunology; D001921:Brain / Q000166:cytology; D002454:Cell Differentiation; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D009836:Oligodendroglia / Q000737:chemistry* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000032:analysis; D014404:Tubulin / Q000032:analysis*
D000911:Antibodies, Monoclonal; D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D014404:Tubulin; C085284:microtubule-associated protein 1B