Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Sep 1;90(3):622–630. doi: 10.1083/jcb.90.3.622

Astroglial cells provide a template for the positioning of developing cerebellar neurons in vitro

PMCID: PMC2111904  PMID: 6793601

Abstract

Indirect immunocytochemical staining with antisera raised against purified glial filament protein and a neurofilament polypeptide was used to study cell interactions between astrocytes and neurons dissociated from embryonic and early postnatal cerebellum. Staining with antibodies raised against purified glial filament protein revealed that greater than 99% of all processes present in cerebellar cultures during the 1st wk in vitro were glial in origin. After 1 wk in culture, unstained processes that were presumably neuronal were observed. Stained astroglial processes formed a dense network that served as a template for cerebellar neurons, identified by indirect immunocytochemical localization of tetanus toxin. More than 90% of neurons from postnatal days 1 or 7 were positioned within one cell diameter of a glial process. In contrast, less than 40% of the neurons dissociated from early embryonic cerebellum were located adjacent to a glial process. Staining with antibodies raised against purified glial filament protein also revealed differences in astroglial morphology that were under developmental regulation. Astroglial cells from embryonic cerebellum were fewer in number and had thick, unbranched processes. Those from postnatal day 1 were more slender, branched, and stellate. Those from postnatal day 7 were highly branched and stellate. Some veil-like astroglial processes were also observed in cells from postnatal animals. These morphological changes were also observed when cells from embryonic day 13 were maintained for a week in vitro. No specific staining of embryonic or postnatal cerebellum cells was observed with antibodies raised against purified neurofilament polypeptides.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Antanitus D. S., Choi B. H., Lapham L. W. Immunofluorescence staining of astrocytes in vitro using antiserum to glial fibrillary acidic protein. Brain Res. 1975 May 23;89(2):363–367. doi: 10.1016/0006-8993(75)90729-5. [DOI] [PubMed] [Google Scholar]
  2. Bignami A., Dahl D. Astrocyte-specific protein and neuroglial differentiation. An immunofluorescence study with antibodies to the glial fibrillary acidic protein. J Comp Neurol. 1974 Jan 1;153(1):27–38. doi: 10.1002/cne.901530104. [DOI] [PubMed] [Google Scholar]
  3. Bignami A., Dahl D. Differentiation of astrocytes in the cerebellar cortex and the pyramidal tracts of the newborn rat. An immunofluorescence study with antibodies to a protein specific to astrocytes. Brain Res. 1973 Jan 30;49(2):393–402. doi: 10.1016/0006-8993(73)90430-7. [DOI] [PubMed] [Google Scholar]
  4. Bignami A., Eng L. F., Dahl D., Uyeda C. T. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res. 1972 Aug 25;43(2):429–435. doi: 10.1016/0006-8993(72)90398-8. [DOI] [PubMed] [Google Scholar]
  5. Del Cerro M., Swarz J. R. Prenatal development of Bergmann glial fibres in rodent cerebellum. J Neurocytol. 1976 Dec;5(6):669–676. doi: 10.1007/BF01181580. [DOI] [PubMed] [Google Scholar]
  6. Eng L. F., Vanderhaeghen J. J., Bignami A., Gerstl B. An acidic protein isolated from fibrous astrocytes. Brain Res. 1971 May 7;28(2):351–354. doi: 10.1016/0006-8993(71)90668-8. [DOI] [PubMed] [Google Scholar]
  7. Fields K. L., Brockes J. P., Mirsky R., Wendon L. M. Cell surface markers for distinguishing different types of rat dorsal root ganglion cells in culture. Cell. 1978 May;14(1):43–51. doi: 10.1016/0092-8674(78)90299-4. [DOI] [PubMed] [Google Scholar]
  8. Hatten M. E. Cell assembly patterns of embryonic mouse cerebellar cells on carbohydrate-derivatized polylysine culture substrata. J Cell Biol. 1981 Apr;89(1):54–61. doi: 10.1083/jcb.89.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hatten M. E., Sidman R. L. Cell reassociation behavior and lectin-induced agglutination of embryonic mouse cells from different brain regions. Exp Cell Res. 1978 Apr;113(1):111–125. doi: 10.1016/0014-4827(78)90092-7. [DOI] [PubMed] [Google Scholar]
  10. Levitt P., Rakic P. Immunoperoxidase localization of glial fibrillary acidic protein in radial glial cells and astrocytes of the developing rhesus monkey brain. J Comp Neurol. 1980 Oct 1;193(3):815–840. doi: 10.1002/cne.901930316. [DOI] [PubMed] [Google Scholar]
  11. Osborn M., Weber K. The display of microtubules in transformed cells. Cell. 1977 Nov;12(3):561–571. doi: 10.1016/0092-8674(77)90257-4. [DOI] [PubMed] [Google Scholar]
  12. Rakic P. Neuron-glia relationship during granule cell migration in developing cerebellar cortex. A Golgi and electronmicroscopic study in Macacus Rhesus. J Comp Neurol. 1971 Mar;141(3):283–312. doi: 10.1002/cne.901410303. [DOI] [PubMed] [Google Scholar]
  13. Rakic P., Sidman R. L. Sequence of developmental abnormalities leading to granule cell deficit in cerebellar cortex of weaver mutant mice. J Comp Neurol. 1973 Nov 15;152(2):103–132. doi: 10.1002/cne.901520202. [DOI] [PubMed] [Google Scholar]
  14. Rakic P., Stensas L. J., Sayre E., Sidman R. L. Computer-aided three-dimensional reconstruction and quantitative analysis of cells from serial electron microscopic montages of foetal monkey brain. Nature. 1974 Jul 5;250(461):31–34. doi: 10.1038/250031a0. [DOI] [PubMed] [Google Scholar]
  15. Schachner M., Hedley-Whyte E. T., Hsu D. W., Schoonmaker G., Bignami A. Ultrastructural localization of glial fibrillary acidic protein in mouse cerebellum by immunoperoxidase labeling. J Cell Biol. 1977 Oct;75(1):67–73. doi: 10.1083/jcb.75.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yen S. H., Fields K. L. Antibodies to neurofilament, glial filament, and fibroblast intermediate filament proteins bind to different cell types of the nervous system. J Cell Biol. 1981 Jan;88(1):115–126. doi: 10.1083/jcb.88.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES