Astrogliosis in culture. IV. Effects of basic fibroblast growth factor

Y. J. Hou, A. C H Yu, J. M R Z Garcia, A. Aotaki-Keen, Y. L. Lee, L. F. Eng, Leonard M Hjelmeland, V. K. Menon

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Abstract

Previous studies have shown that the mechanical wounding of 3-week-old cultured rat astrocytes results in cell proliferation and hypertrophy resembling astrocyte responses to a brain injury in vivo. We now report the effects of basic fibroblast growth factor (bFGF) and an anti-bFGF antibody on astrocyte morphology, proliferation, and migration following in vitro wounding of confluent secondary cultures. Addition of bFGF (20 ng/ml) to wounded cultures induced morphological changes characteristic of differentiation in wounded and nonwounded areas of the culture. Combined treatment with bFGF and an anti-bFGF antibody (100 μg/ml) prevented this effect. Astrocyte proliferation along the edges of a scratch wound was at maximum 24 hr after wounding in cells growing in Eagle's minimum essential medium (EMEM) containing 10% serum. Low serum concentration and treatment with dibutyryl cyclic adenosine monophosphate (dbc-AMP) reduced injury- associated astrocyte proliferation. Addition of bFGF to cultures in EMEM with serum increased astrocyte proliferation at 18 and 24 hr after wounding. This effect was reduced considerably by treatment of cultures with bFGF in combination with an anti-bFGF antibody. The combined treatment and the antibody alone reduced cell division to a level lower than in control cultures. Twenty-four hr following wounding, astrocytes along the edges of the wound exhibited extension of thick, flat processes into the wound area. At 3 and 5 days after wounding, a bodily migration of astrocytes into the wounded area was observed. Addition of bFGF significantly increased astrocyte migration 1 day after wounding, with maximum effect on day 3 and no subsequent increase on day 5. A combination of bFGF and anti-bFGF antibody as well as the antibody alone reduced astrocyte migration to a level lower than in controls. Immunohistochemical localization and isoform pattern of bFGF in astrocytes did not change with dbc-AMP treatment or wounding. We conclude that mechanically wounded confluent astrocytes respond to bFGF added to the culture medium by enhancing cell division, differentiation, and migration. In addition, the results of the antibody treatment also suggest a role for endogenous bFGF in astrocyte proliferation and migration elicited by wounding in vitro. These results support the notion that in vivo, both bFGF released by injury and endogenous bFGF synthesized by astrocytes, contribute to the cellular responses that lead to astrogliosis.

Original languageEnglish (US)
Pages (from-to)359-370
Number of pages12
JournalJournal of Neuroscience Research
Volume40
Issue number3
DOIs
StatePublished - 1995

Keywords

  • astrogliosis
  • culture
  • glia
  • trauma

ASJC Scopus subject areas

  • Neuroscience(all)

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    Hou, Y. J., Yu, A. C. H., Garcia, J. M. R. Z., Aotaki-Keen, A., Lee, Y. L., Eng, L. F., Hjelmeland, L. M., & Menon, V. K. (1995). Astrogliosis in culture. IV. Effects of basic fibroblast growth factor. Journal of Neuroscience Research, 40(3), 359-370. https://doi.org/10.1002/jnr.490400310