Abstract
We have grown neonatal mouse astrocytes (2 days old) in cell culture for nine days followed by mechanical removal of the contaminating oligodendrocytes. Astrocytes are continued in culture in serum-free medium for an additional 5 days. Assays for both glutamine synthetase and glial fibrillary acidic protein revealed the cultures to contain predominantly astrocytes. Cell pellet lysates and the glial conditioned medium (GCM) from these cultures were assayed for βNGF synthesis and secretion by radioimmunoassay (RIA) using βNGF purified from male mouse submaxillary gland as standard. Biosynthetic labeling with S35-cystine labelled at 24 hour intervals for 120 hours demonstrated cellular synthesis of NGF between 72 and 96 hours with secretion of labelled NGF into the GCM between 96 and 120 hours. Immunoprecipitates from cell homogenate supernatants and GCM on polyacrylamide gel electrophoresis (PAGE) showed bands in the same molecular weight range as purified βNGF. Partial purification of the NGF-like material in the GCM with Sephadex G-100 gel filtration showed both immunoreactivity and bioactivity; bioactivity was blocked by anti-NGF antiserum.
Results and conclusions: 1) An NGF-like factor (NGF-LF) is synthesized and secreted by neonatal mouse astrocytes in culture, 2) NGF-LF is similar, if not identical to βNGF synthesized in the mouse submaxillary gland, 3) NGF-LF is produced locally in normal brain tissue, 4) NGF-LF may be important in the developing CNS.
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Weichsel, M., Tarris, R. & Fisher, D. 332 IN VITRO SYNTHESIS AND SECRETION OF A NERVE GROWTH FACTOR (NGF) BY NEONATAL MOUSE ASTROCYTES. Pediatr Res 19, 166 (1985). https://doi.org/10.1203/00006450-198504000-00362
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DOI: https://doi.org/10.1203/00006450-198504000-00362