Expression of the immunoglobulin superfamily cell adhesion molecule F3 by oligodendrocyte-lineage cells.
We have analysed the expression of glycosylphosphatidylinositol (GPI)-anchored proteins by oligodendrocyte-lineage cells. Biosynthetic labeling of mouse oligodendroglial primary cultures and an oligodendroglial precursor cell line demonstrated that these cells synthesise a variety of different GPI-anchored proteins. GPI-anchored proteins were isolated as a bulk preparation from the precursor cell line, and the individual proteins separated by 2D gel electrophoresis and analysed by microsequencing after tryptic digestion of the separated components. One of the most prominent GPI-anchored proteins synthesised by the cell line was identified as the cell adhesion molecule F3, previously thought to be exclusively expressed by neurons. Western blotting and immunoprecipitation with several polyclonal sera confirmed the expression of F3 by oligodendrocyte-lineage cells and demonstrated the presence of F3 in myelin. Double staining with a panel of oligodendrocyte-specific antibodies and anti-F3 antibodies of cerebellar cultures, as well as oligodendrocytes isolated by panning, showed a colocalization of F3 with oligodendrocyte markers. Oligodendrocyte F3 is shown to be susceptible to phosphatidylinositol-phospholipase C (PI-PLC) cleavage, similar to neuronal F3. Northern blots demonstrated that the oligodendroglial F3 mRNA is the same size as the neuronal message; however, no F3 mRNA could be detected in cortical astrocytes and an astrocytic cell line. Thus, in addition to the expression by neurons, the cell-type specificity of F3 expression must be extended to oligodendroglial cells, underscoring the importance of this Ig superfamily member in the nervous system.
Koch
T
T
Department of Neurobiology, University of Heidelberg, Germany.
Brugger
T
T
Bach
A
A
Gennarini
G
G
Trotter
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D015152:Blotting, Northern; D015153:Blotting, Western; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism*; D019070:Cell Lineage; D002478:Cells, Cultured; D058966:Contactins; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D051379:Mice; D009185:Myelin Proteins / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D011817:Rabbits
D000911:Antibodies, Monoclonal; D015816:Cell Adhesion Molecules, Neuronal; D058966:Contactins; D017261:Glycosylphosphatidylinositols; D009185:Myelin Proteins; D011506:Proteins