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Related Experiment Videos

Fibroblast growth factors decrease inducible nitric oxide synthase mRNA accumulation in bovine retinal pigmented

O Goureau1, V Faure, Y Courtois

  • 1Laboratoire de Recherches Gérontologiques, Institut National de la Santé et de la Recherche Médicale, Paris, France.

European Journal of Biochemistry
|June 15, 1995
PubMed
Summary
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Fibroblast growth factors (FGF) inhibit inducible nitric oxide (NO) synthase expression in bovine retinal pigmented epithelial (RPE) cells. This FGF-mediated reduction in NO synthase may protect the retina from inflammatory damage.

Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Immunology

Background:

  • Bovine retinal pigmented epithelial (RPE) cells express inducible nitric oxide (NO) synthase upon activation with interferon gamma (IFN-γ) and lipopolysaccharide (LPS).
  • Fibroblast growth factors (FGF), specifically FGF-1 and FGF-2, are known inhibitors of this NO synthase activity.

Purpose of the Study:

  • To elucidate the molecular mechanisms by which FGF inhibits NO synthase activity in RPE cells.
  • To investigate the effect of FGF on the expression and stability of inducible NO synthase mRNA.

Main Methods:

  • Immunocytochemistry and Western blot analysis to quantify NO synthase protein levels.
  • Northern blot analysis using a mouse macrophage NO synthase cDNA probe to assess mRNA levels.
  • Message stability studies to determine the impact of FGF on mRNA degradation.

Related Experiment Videos

  • Assessment of IFN-γ receptor binding in the presence of FGF.

    • Main Results:

    • FGF-2 co-stimulation with LPS/IFN-γ resulted in lower NO synthase accumulation in RPE cells.
    • FGF-1 and FGF-2 significantly reduced the level of inducible NO synthase mRNA in LPS/IFN-γ-activated RPE cells.
    • FGF did not affect mRNA degradation, suggesting regulation at the expression level.
    • FGF did not alter IFN-γ binding to its receptors.

    Conclusions:

    • FGF inhibits inducible NO synthase expression in RPE cells, likely at the transcriptional level.
    • FGF does not appear to act by altering mRNA stability or IFN-γ receptor binding.
    • FGF's inhibition of NO synthase may offer retinal protection against inflammatory and endotoxin-mediated damage.