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Dexamethasone Inhibits Growth Factor-Induced Type 3 Deiodinase Activity and mRNA Expression in a Cultured Cell Line Derived from Rat Neonatal Brown Fat Vascular-Stromal Cells

Departments of Medicine and Physiology, Dartmouth Medical School, Lebanon, New Hampshire 03756

Address all correspondence and requests for reprints to: Donald L. St. Germain, Departments of Medicine and Physiology, Dartmouth Medical School, One Medical Center Drive, Lebanon, New Hampshire 03756. E-mail: . stgermain{at}dartmouth.edu

Studies examining the regulation of the type 3 deiodinase (D3) have been hampered by the lack of cell lines that constitutively express this enzyme. To address this issue, a new cell line, designated brown fat vascular-stromal (BVS-1), was generated by continuous subculturing of precursor cells derived from the vascular-stromal fraction of rat neonatal brown fat. BVS-1 cells did not differentiate into adipocytes when cultured for 5 d in DMEM supplemented with 2% newborn calf serum, 4 nM insulin, 2 nM T₃, and 10 nM dexamethasone (DEX). However, when cultured in regular medium, the cells expressed high levels of D3 activity (1–5 pmol/h per milligram protein) and mRNA. D3 mRNA was markedly induced by treatment for 6 h with epidermal growth factor, acid or basic fibroblast growth factors (10 ng/ml), or a 3-h treatment with a phorbol ester [12-O-tetradecanoylphorbol-13-acetate (TPA), 1 µM] or 10% fetal bovine serum. However, preincubation of cells overnight with 50 nM DEX completely blocked the D3-inducing effects of basic fibroblast growth factor. The DEX effect was partially blocked when a glucocorticoid receptor antagonist was present. Overnight DEX treatment (50 nM) also decreased basal D3 activity by 80%. In summary, we have established BVS-1 cells as a continuous cell line useful for studying the regulation of D3 expression. Furthermore, we have shown that DEX inhibits growth factor-induced D3 expression in these cells.

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