Glucocorticoid receptors and inhibition of neonatal mouse dermal fibroblast growth in primary culture.
Primary cultures of dermal fibroblasts from neonatal mice were used to investigate some of the anti-inflammatory effects of glucocorticoids in vitro as influenced by the genetic background of two different strains of mice (A/J and C47 Bl/6J). Fibroblasts were cultured in the absence or presence of various glucocorticoids for 2-7 days. After 4-7 days in the presence of steroid, DNA synthesis was reduced by 50-85% while protein synthesis was inhibited by 50-60%. Corticosterone produced a dose-dependent inhibition of DNA synthesis in these cells with a 50% reduction occurring at 10 nM. Specific, high affinity, low capacity binding proteins for [3H]dexamethasone or [3H]triamcinolone acetonide were identified in the cytoplasm of neonatal dermal fibroblasts which had an apparent Kd of 9 nM and approximately 5,200-6,400 binding sites/cell. Sedimentation analysis of the [3H]triamcinolone acetonide-receptor complexes on low salt glycerol gradients exhibited binding in the 7 to 8 S region of the gradients. These studies demonstrate that inhibition of growth of primary cultures of mouse neonatal dermal fibroblasts by glucocorticoids is probably mediated by a receptor-mediated pathway, and that this primary culture system might be useful in delineating other anti-inflammatory effects of glucocorticoids in vitro.
D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D003907:Dexamethasone / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D051379:Mice; D014176:Protein Biosynthesis; D011965:Receptors, Glucocorticoid / Q000187:drug effects*; D011987:Receptors, Steroid / Q000187:drug effects*; D012867:Skin / Q000166:cytology*
D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D003907:Dexamethasone; D004247:DNA