Steroids and Triiodothyronine Reduce Nerve Growth Factor Concentrations in Medium Conditioned by L-929 Fibroblasts*

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Abstract
Thyroid hormone, testosterone, and glucocorticoids stimulate the production of nerve growth factor (NGF) in mouse submandibular glands, but little is known about the influences of these hormones on other sources of NGF. In this study we tested the effects of thyroid hormone and steroids on NGF levels in medium conditioned by L-929 fibroblasts (Lcells), a mouse cell line that secretes NGF in culture. NGF concentrations in conditioned medium were measured by RIA and were normalized for cell number. In the absence of hormones, NGF levels in medium averaged 5.57 ng/ml. T3, cortisone, and aldosterone decreased NGF levels below the sensitivity of the RIA (0.37 ng/ml), with respective ID50 values of 1 × 10M-5, 1 × 10-8, and 5 × 10-8 M. Testosterone decreased NGF levels by 66%, with an IDso of 9 × 10-9 M, and 17β-estradiol decreased it by up to 50%, although the hormone′s effects had not plateaued at 10-6 M. NGF concentrations were unaffected by progesterone at doses as high as 10-6 M. Cell viability was unaffected by hormone treatment, and the total amount of secreted protein in medium decreased only in cultures treated with high doses of cortisone and aldosterone. Qualitatively similar results were obtained by bioassay; steroids markedly reduced, but did not eliminate, the nerve growth-promoting effects of L-cells on embryonic chick dorsal root ganglia in cocultures. These data indicate that hormones that stimulate NGF production in salivary glands inhibit NGF production by L-cells in culture. (Endocrinology118: 1417-1425, 1986)