The Cytoskeleton of Primary Astrocytes in Culture Contains Actin, Glial Fibrillary Acidic Protein, and the Fibroblast‐Type Filament Protein, Vimentin

The Cytoskeleton of Primary Astrocytes in Culture Contains Actin, Glial Fibrillary Acidic... Abstract: Primary astrocytes were cultured from the forebrains of 1‐day‐old rats. Immunofluorescence microscopy showed that approximately 80% of the cells were positive for glial fibrillary acidic protein (GFAP) and >80% were stained with an antiserum to the molecular weight 58,000 fibroblast intermediate filament protein (vimentin). Gel electrophoresis of Triton‐insoluble cytoskeleton preparations from these cultures revealed three major bands having molecular weights of 58,000, 51,000, and 42,000, together with some prominent lower‐molecular‐weight species. The protein of molecular weight 51,000 was not present in preparations from fibroblasts. Each of the three major astrocyte proteins was subjected to limited proteolysis, while two of the proteins were cleaved by cyanogen bromide. The electrophoretic peptide patterns of the 58,000 protein were similar to those of vimentin isolated from NIL‐8 fibroblasts, and the patterns of the 51,000 protein were similar to those of GFAP isolated from rat spinal cord. The patterns of the protein of molecular weight 42,000 resembled those of muscle actin. Rocket immunoelectrophoresis showed that the 51,000 astrocyte protein reacted with an antiserum to bovine GFAP, but the 58,000 and 42,000 proteins failed to react. We conclude that the major proteins of cytoskeleton preparations from cultured primary astrocytes are vimentin (58,000), GFAP (51,000), and actin (42,000), and that our data show no obvious structural relationship among them. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neurochemistry Wiley

The Cytoskeleton of Primary Astrocytes in Culture Contains Actin, Glial Fibrillary Acidic Protein, and the Fibroblast‐Type Filament Protein, Vimentin

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Publisher
Wiley
Copyright
Copyright © 1981 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0022-3042
eISSN
1471-4159
D.O.I.
10.1111/j.1471-4159.1981.tb05302.x
Publisher site
See Article on Publisher Site

Abstract

Abstract: Primary astrocytes were cultured from the forebrains of 1‐day‐old rats. Immunofluorescence microscopy showed that approximately 80% of the cells were positive for glial fibrillary acidic protein (GFAP) and >80% were stained with an antiserum to the molecular weight 58,000 fibroblast intermediate filament protein (vimentin). Gel electrophoresis of Triton‐insoluble cytoskeleton preparations from these cultures revealed three major bands having molecular weights of 58,000, 51,000, and 42,000, together with some prominent lower‐molecular‐weight species. The protein of molecular weight 51,000 was not present in preparations from fibroblasts. Each of the three major astrocyte proteins was subjected to limited proteolysis, while two of the proteins were cleaved by cyanogen bromide. The electrophoretic peptide patterns of the 58,000 protein were similar to those of vimentin isolated from NIL‐8 fibroblasts, and the patterns of the 51,000 protein were similar to those of GFAP isolated from rat spinal cord. The patterns of the protein of molecular weight 42,000 resembled those of muscle actin. Rocket immunoelectrophoresis showed that the 51,000 astrocyte protein reacted with an antiserum to bovine GFAP, but the 58,000 and 42,000 proteins failed to react. We conclude that the major proteins of cytoskeleton preparations from cultured primary astrocytes are vimentin (58,000), GFAP (51,000), and actin (42,000), and that our data show no obvious structural relationship among them.

Journal

Journal of NeurochemistryWiley

Published: Jul 1, 1981

References

  • Intermediate filaments from bovine, rat, and human CNS: Mapping analysis of the major proteins
    Chiu, Chiu; Korey, Korey; Norton, Norton
  • Isolation and characterization of glial filaments from human brain
    Goldman, Goldman; Schaumburg, Schaumburg; Norton, Norton
  • Kinetics of adenosine uptake into astrocytes
    Hertz, Hertz
  • Filament organization revealed in platinum replicas of freeze‐dried cytoskeletons
    Heuser, Heuser; Kirschner, Kirschner
  • Ultrastructural localization of glial fibrillary acidic protein in mouse cerebellum by immunoperoxidase labelling
    Schachner, Schachner; Hedley‐Whyte, Hedley‐Whyte; Hsu, Hsu; Schoonmaker, Schoonmaker; Bignami, Bignami
  • Antibodies to neurofilament, glial filament and fibroblast intermediate filament proteins bind to different cell types of the nervous system
    Yen, Yen; Fields, Fields

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