Cell blotting and isoelectric focusing of neuroblastoma-derived heparin-binding neurotrophic activities: detection of basic fibroblast growth factor protein and mRNA.
Neuroblastoma cells have been shown to contain growth factors, which may be involved in the regulation of cell proliferation and/or differentiation. We have examined whether human IMR-32 neuroblastoma cells store factors with a capacity to promote neuron survival and differentiation. Heparin affinity chromatography, cell blotting, and isoelectric focusing of IMR-32 cell extracts revealed multiple neurotrophic activities at molecular weights of 16.8, 24.1, 39.0, 45.3, 52.3 and isoelectric points of 4.0, 5.0, 8.3, 9.0. Immunocytochemistry, immunoblotting, and radio-immunoassay with specific antibodies suggests that one neurotrophic activity is immunologically identical with basic fibroblast growth factor (bFGF). This assumption is supported by Northern blot analysis, which shows a 6.0 kb bFGF transcript.
Wewetzer
K
K
Department of Anatomy and Cell Biology, University of Heidelberg, Germany.
Janet
T
T
Heymann
D
D
Unsicker
K
K
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology; D002642:Chick Embryo; D002846:Chromatography, Affinity; D016207:Cytokines / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000276:immunology / Q000378:metabolism*; D005726:Ganglia, Parasympathetic / Q000166:cytology; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007525:Isoelectric Focusing; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000302:isolation & purification; D011863:Radioimmunoassay; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D016207:Cytokines; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; C066845:pleiotrophin; D006493:Heparin