Spontaneous morphological changes by overexpression of the growth-associated protein B-50/GAP-43 in a PC12 cell line

In order to study the direct effects of B-50 on neural cell morphology, rat B-50 cDNA was transfected into a PC12 cell line (PC-B2) exhibiting neurite outgrowth independent of the expression of endogenous B-50. The morphological changes were visualized by confocal scanning laser microscopy using fluorescence labelling for B-50 and for F-actin. The transfected cells exhibited filopodia and/or blebs on the plasma membrane, containing most of the B-50 immunoreactivity. No spontaneous neurite outgrowth was observed. Following NGF treatment transfected and nontransfected PC-B2 cells extended F-actin positive filopodia and neurites with a striking colocalisation of B-50 and F-actin. Our data show that the presence of B-50 can influence cell surface morphology independent of the presence of NGF. The colocalisation of B-50 and F-actin in the filopodial protrusions but not in the blebs might be indicative for a role of B-50 in actin polymerization and depolymerization.