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Epidermal growth factor (EGF) stimulation of cultured brain cells. II. Increased production of extracellular soluble proteins.
Developmental Brain Research
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The production of extracellular soluble proteins was studied in serum-free aggregating cell cultures of fetal rat telencephalon labeled on culture day 7 with a mixture of radioactive amino acid precursors. Cultures treated continuously with epidermal growth factor (EGF; 20 ng/ml) showed a generally increased protein secretion and a particularly enhanced production of a few distinct extracellular proteins. The time lag of this response after an initial dose of EGF (25 ng/ml) on day 7 was 48 h. The total macromolecular radioactivity that accumulated within 96 h of labeling in the media of EGF-treated cultures was 175% of untreated controls, whereas no difference was found in the proportions of intracellular amino acid incorporation. Cultures which received a single dose of EGF (25 ng/ml) on day 1 showed still a greatly increased protein secretion on day 7. Prevention of extracellular protein accumulation by reducing the initial cell number and increasing the rate of media changes did not affect the EGF-induced stimulation of the two glial enzymes, glutamine synthetase and 2',3'-cyclic nucleotide 3'-phosphohydrolase. The results suggest that both the increased production of extracellular proteins and the enhanced development of glial enzymatic activities reflect the stimulated phenotypic expression of EGF-sensitive brain cells.
2',3'-Cyclic Nucleotide 3'-Phosphodiesterase, 2',3'-Cyclic-Nucleotide Phosphodiesterases/metabolism, Animals, Cell Aggregation, Cells, Cultured, Epidermal Growth Factor/pharmacology, Fetus, Glutamate-Ammonia Ligase/metabolism, Nerve Tissue Proteins/biosynthesis, Neuroglia/enzymology, Phosphoric Diester Hydrolases, Rats, Rats, Inbred Strains, Telencephalon/metabolism
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