Expression of basic fibroblast growth factor and its receptors in human fetal microglia cells

Abstract

The presence of basic fibroblast growth factor (bFGF) and FGF receptors was investigated in microglia cells derived from human fetal brain long-term cultures. Production of bFGF was suggested through the capability of microglial extracts to stimulate plasminogen activator (PA) synthesis in endothelial cells. The identity of PA-stimulating activity with bFGF was confirmed by its high affinity for heparin and its cross-reactivity with polyclonal antibodies to human recombinant bFGF. These antibodies recognized a cell-associated Mr 18,000 protein as well as trace amounts of the Mr 24,000 bFGF isoform in Western blot. All microglial cells showed bFGF immunoreactivity in the cytoplasm and, sometimes, in the nucleus.Scatchard plot analysis of 125I-bFGF binding data revealed the presence of low affinity heparan-sulphate proteoglycans (380,000±60,000 sites/cell; Kd = 30±9 pM). Immunocytochemistry confirmed the presence of FGF receptors (10,300+2500 sites/cell; Kd = 30±9 pM). Immunocytochemistry confirmed the presence of FGF receptor (1/flg) on the cell surface of some, but not all microglial cells, with prevalent association to ameboid microglia. Transcripts for FGF receptors 1,2,3 and 4 were found in microglia by Northern blot analysis.Co-expression of bFGF and its receptors in human fetal microglia suggests an autocrine role of bFGF in these cells.