Spatiotemporal gradient of oligodendrocyte differentiation in chick optic tectum requires brain integrity and cell-cell interactions

Abstract

The development of oligodendrocytes in the chicken optic tectum (OT) was studied in vivo and in vitro by analyzing expression of myelin-associated glycoprotein (MAG) with a monoclonal antibody. MAG+ cells first appeared in the anterior OT on embryonic day (E) 12, were present throughout the anterior half on E15, and eventually filled the tectum on E17. This spatiotemporal appearance of MAG+ oligodendrocytes resembled two streams of cells entering the OT along the afferent and efferent axonal layers. However, experiments determined that this appearance of MAG immunoreactivity was the result of a gradient of oligodendrocyte differentiation and was not cell migration. First, retroviral vector labeling of OT progenitors in vivo on E3 resulted in labeled oligodendrocytes in late embryos. In addition, pieces of OT from as early as E3 kept in culture for a week developed numerous MAG+ oligodendrocytes. Pieces of both anterior and posterior E7 OT developed MAG+ oligodendrocytes after 3 days in culture, well ahead of their normal schedule in vivo. BrdU incorporation studies revealed that these cells were not born in culture, but merely differentiated. Monolayer cultures made from dissociated E10 or later OT cells developed MAG+ oligodendrocytes, but monolayers made from E7 OT cells did not. These experiments demonstrate that oligodendrocyte progenitors were present in the OT as early as E3, that they could differentiate precociously, and that their normal progressive differentiation in situ must be due to removal of inhibitory constraints rather than the onset of inductive factors. Also, certain cell-cell interactions occur between E7 and E10, which cannot be disrupted if oligodendrocyte differentiation is to occur. © 2003 Wiley-Liss, Inc.