Suppression by antisense mRNA demonstrates a requirement for the glial fibrillary acidic protein in the formation of stable astrocytic processes in response to neurons.
Open Access
- 15 March 1991
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 112 (6), 1205-1213
- https://doi.org/10.1083/jcb.112.6.1205
Abstract
The glial fibrillary acidic protein (GFAP) is a glial-specific intermediate filament protein, which is expressed in astrocytes in the central nervous system, as well as in astrocytoma cell lines. To investigate the function of GFAP, we have studied the human astrocytoma cell line, U251, which constitutively expresses GFAP and vimentin in the same 10-nm filaments. These cells respond to neurons in vitro in the same way as primary astrocytes: they withdraw from the cell cycle, support neuronal cell survival and neurite outgrowth, and they extend complex, GFAP-positive processes. To determine the role of GFAP in these responses, we have specifically suppressed its expression by stably transfecting the U251 cells with an antisense GFAP construct. Two stable antisense cell lines from separate transfections were isolated and were shown to be GFAP negative by Northern and Western blot analyses, and by immunofluorescence studies. The antisense cell lines were inhibited in their ability to extend significant glial processes in response to neurons. In culture with primary neurons, the average increase in process length of the U251 cells was nearly 400%, as compared to only 14% for the antisense transfectants. The other neuron induced responses of astrocytes, i.e., proliferative arrest and neuronal support, were not affected in these cell lines. These data support the conclusion that the glial-specific intermediate filament protein, GFAP, is required for the formation of stable astrocytic processes in response to neurons.Keywords
This publication has 21 references indexed in Scilit:
- C17, a retrovirally immortalized neuronal cell line, inhibits the proliferation of astrocytes and astrocytoma cells by a contact‐mediated mechanismGlia, 1990
- Sequence of a cDNA clone encoding mouse glial fibrillary acidic protein: structural conservation of intermediate filaments.Proceedings of the National Academy of Sciences, 1984
- Identification of glial filament protein and vimentin in the same intermediate filament system in human glioma cells.Proceedings of the National Academy of Sciences, 1984
- Development of cerebellar astroglia: Transitions in form and cytoskeletal contentDevelopmental Biology, 1984
- VIMENTIN, THE 57000 MOLECULAR-WEIGHT PROTEIN OF FIBROBLAST FILAMENTS, IS THE MAJOR CYTOSKELETAL COMPONENT IN IMMATURE GLIA1981
- Immunofluorescence localization of HeLa cell microtubule-associated proteins on microtubules in vitro and in vivo.The Journal of cell biology, 1980
- Selective iodination and polypeptide composition of pinocytic vesicles.The Journal of cell biology, 1980
- DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells.Proceedings of the National Academy of Sciences, 1979
- Preparative and analytical purification of DNA from agarose.Proceedings of the National Academy of Sciences, 1979
- Mode of cell migration to the superficial layers of fetal monkey neocortexJournal of Comparative Neurology, 1972