Site selectivity of osteoblast gene expression response to thyroid hormone localized by in situ hybridization

We have previously reported that thyroid-stimulating hormone (TSH)-suppressive doses of L-thyroxine (L-T4) decrease femoral, but not vertebral, bone mineral density (BMD) in rats. L-T4-induced decreases in BMD were associated with increased expression of genes, reflecting osteoblast activity in mRNA extracted from whole femurs but not from vertebrae. To document that this skeletal selectivity reflected altered osteoblast activity, we studied gene expression by in situ hybridization in 8-wk-old rats treated with L-T4 (20 microg x 100 g body wt(-1) x day(-1)) for 4 wk. TSH-suppressive doses of L-T4 were associated with decreased femoral (0.299 +/- 0.005 vs. 0.273 +/- 0.005 g/cm2, P < 0.01), but not vertebral (0.222 +/- 0.004 vs. 0.218 +/- 0.003 g/cm2), BMD. In situ hybridization documented that L-T4 administration for 4 wk increased expression of osteocalcin and alkaline phosphatase mRNA in femoral, but not vertebral, osteoblasts. This study demonstrates a differential gene expression response of vertebral and femoral osteoblasts to L-T4. This altered degree of gene expression markers of osteoblast activity documented by in situ hybridization may in part explain the apparent clinical differences in the effect of L-T4 on femoral and vertebral BMD.