Summary
The capability of epidermal growth factor (EGF) to stimulate DNA synthesis in neural cells was investigated in primary cultures of early postnatal mouse cerebellum. At concentrations of 10-8M, EGF stimulates DNA synthesis in astrocytes, which were identified immunocytologically by the cell type-specific marker, glial fibrillary acidic (GFA) protein. Astrocytes express cell-surface receptors for EGF as can be shown by binding of [125I]-labeled EGF to live monolayer cultures. In the presence of 10% horse serum, EGF stimulates DNA-synthesis by a factor of about two-fold. Stimulation by EGF over control values is approximately 4-fold in the presence of 1% serum and 6 to 10-fold in the absence of serum. Absolute numbers of astrocytes are increased after more prolonged action of EGF. DNA-synthesis in neurons or oligodendroglia is not significantly stimulated by EGF. EGF enhances cell survival of serum-deprived cerebellar cultures. Fibroblast growth factor does not increase DNA-synthesis in astrocytes under the conditions used in this study.
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Abbreviations
- BME :
-
basal medium, Eagle's
- BME-BSA :
-
basal medium, Eagle's containing 0.1% bovine serum albumin
- EDTA :
-
ethylene-diamino-N, N′-tetraacetic acid
- EGF :
-
epidermal growth factor
- FGF :
-
fibroblast growth factor
- GFA :
-
glial fibrillary acidic
- HS :
-
horse serum
- [3 H] TdR:
-
tritium-labeled thymidine
- PAP :
-
peroxidase-anti-peroxidase
- PBS :
-
phosphate-buffered saline
- SDS :
-
sodium dodecyl sulfate
- TCA :
-
trichloroacetic acid
References
Bignami A, Dahl D (1974) Astrocyte specific protein and neuroglial differentiation. An immunofluorescent study with antibodies to GFAP. J Comp Neurol 153:27–38
Brockes JP, Lemke GE (1981) The neuron as a source of mitogen; its influence on the proliferation of glial and non-neural cells. Society for Developmental Biology Symposium V-“Development in the Nervous System” Feldman JD, Garrod DR (eds) Cambridge Univ Press, in press
Carpenter G, Cohen S (1979) Epidermal growth factor. Ann Rev Biochem 48:193–216
Fields KL, Brockes JP, Mirsky R, Wendon LBM (1978) Cell Surface markers for distinguishing different types of rat DRG cells Cell 4:43–51
Gospodarowicz D, Greenburg G, Bialecki H, Zetter BR (1978) Factors involved in the modulation of cell proliferation in vivo and in vitro: The role of fibroblast and epidermal growth factors in the proliferative response of mammalian cells. In vitro 14:85–118
Gospodarowicz D, Moran JS (1976) Growth factors in mammalian cell culture. Ann Rev Biochem 44:531–559
Gospodarowicz D, Moran JS, Mescher AL (1976) Cellular Specifities to Fibroblast Growth Factor and Epidermal Growth Factor. The Thirty-Fifth Symposium of the Society for Developmental Biology. Academic Press, Inc. N.Y., 1978
Gray WR (1967) Dansyl chloride procedure. In: Hirs CHW (ed) Meth Enzym, Vol 11. Academic Press N.Y., pp 139–151
Heldin CH, Westermark B, Wasteson A (1979) Desensitisation of cultured glial cells to epidermal growth factor by receptor down-regulation. Nature 282:419–420
Lagenaur C, Sommer, I, Schachner M (1980) Subclass of astroglia in mouse cerebellum recognized by monoclonal antibody. Dev Biol 79:367–378
Ponten J, Macintyre EH (1968) Long term culture of normal and neoplastic human glia. Acta Path Microbiol Scand 74:465–486
Savage RC Jr, Cohen S (1972a) Epidermal growth factor and a new derivative. J Biol Chem 247:7609–7611
Savage RC Jr, Juagami T, Cohen S (1972b) The primary structure of epidermal growth factor. J Biol Chem 247:7612–7621
Schachner M, Willinger M (1979) Cell type specific cell surface antigens in the cerebellum. Progress in Brain Res 51:23–44
Schachner M, Hedley-Whyte ET, Hsu DW, Schoonmarker G, Bigami A (1977) Ultrastructural localisation of GFAP in mouse cerebellum by immunoperoxidase labeling. J Cell Biol 75:67–73
Schachner M, Kim SK, Zehnle R (1981) Developmental expression in central and peripheral nervous system of oligodendrocyte cell surface antigens (0 antigens) recognized by monoclonal antibodies. Dev Biol 83:328–338
Schnitzer J, Schachner M (1981) Expression of Thy-1, H-2 and NS-4 cell surface antigens and tetanus toxin receptors in developing and adult mouse cerebellum. J Neuroimmunol, in press
Smart I, Leblond CP (1961) Evidence for division and transformation of neuroglia cells in the mouse brain as derived from radioautography after injection of thymidine — 3H. J Comp Neurol 116:349–367
Sommer I, Schachner M (1981) Monoclonal antibodies (01 to 04) to oligodendrocyte cell surfaces: an immunocytological study in the central nervous system. Dev Biol 83:311–327
Sternberger L (1979) Immunocytochemistry. John Wiley and Sons, Inc NY (Second edition)
Vaheri A, Ruoslathi E, Westermark B, Ponten J (1976) A common cell-type specific surface antigen in cultured human glial cells and fibroblasts: loss in malignant cells. J Exp Med 143:64–72
Wand M, Zenthen E, Evans EA (1967) Tritiated thymidine: effect of decompostition by self radiolysis on specificity as a tracer for DNA synthesis. Science 157:436–438
Westermark B (1976) Density dependent proliferation of human glia cells stimulated by epidermal growth factor. BBRC 69:304–310
Westermark B, Wasteson A (1975) The response of cultured human and normal glia cells to growth factors. Adv Metab Disorb 8:85–100
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Leutz, A., Schachner, M. Epidermal growth factor stimulates DNA-synthesis of astrocytes in primary cerebellar cultures. Cell Tissue Res. 220, 393–404 (1981). https://doi.org/10.1007/BF00210517
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DOI: https://doi.org/10.1007/BF00210517