Expression of neuromodulin (GAP-43) and its regulation by basic fibroblast growth factor during the differentiation of O-2A progenitor cells.

Abstract

In a recent work we have shown that neuromodulin (Nm, also known as GAP-43), a protein kinase C substrate, previously believed to be expressed exclusively in neurons, is also present in glial cells. Here we investigated the expression of Nm and its mRNA in O-2A glial progenitor cells (common precursor for oligodendrocytes and type-2 astrocytes) during their development in secondary culture and under the influence of basic fibroblast growth factor (bFGF). The different stages of oligodendrocyte development were characterized by the expression of surface markers: A2B5, which identifies O-2A glial precursor cells, and O4 and galactocerebroside (GC), which characterize later developmental stages. The number of cells expressing Nm (about 90% at culture initiation) decreased rapidly during the first 2 days and reached a plateau at around 30-40%. The level of Nm mRNA followed a similar kinetic. Immunocytochemistry demonstrated that at 4 days in vitro about 25-30% cells were A2B5+, 30-40% Nm+, a high percentage (60-70%) O4+, and 35-40% GC+. Nearly all of the morphologically immature A2B5+ cells expressed also the Nm antigen, very few of the O4+ cells still expressed Nm and almost no cells expressed both GC and Nm. Most O4+ cells developed a typical oligodendrocyte morphology and were essentially GC+. This study also showed that in the presence of serum, the A2B5+ Nm+ and O4+ Nm+ (GC-) cells retained their bipotentiality and differentiated into GFAP+ (glial fibrillary acidic protein) Nm+ type-2 astrocytes. The bFGF was found to stimulate the proliferation of Nm+ 0-2A precursor cells and to increase the level of Nm mRNA. At 4 days under this culture condition, the predominant cell type was A2B5+ and Nm+. Only 25-35% of the cells were O4+, but 90-95% of them were Nm+. Very few GC+ cells were visible in the presence of bFGF, but 20-40% of them were Nm+. These data indicate that Nm is essentially associated to glial O-2A precursor cells and further confirm that bFGF blocks the differentiation of these cells. It is suggested that Nm plays a role in the plasticity (developmental potential) of the bipotential 0-2A progenitor cells.